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Anxiolytic-like effect of succinic acid in mice


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The putative anxiolytic activity of succinic acid was examined in male mice by using a number of experimental paradigms of anxiety and compared with that of the known anxiolytic compound diazepam. Use of the elevated plus-maze test revealed that diazepam (1.0, 2.0 and 4.0 mg/kg, PO) or succinic acid (3.0 or 6.0 mg/kg, PO) increased the percentage of entries into open arms and of time spent on open arms. In novel food consumption test, succinic acid (3.0, 6.0, and 12.0 mg/kg, IP) caused significant increases in food intake during 5 min when compared with the vehicle. In the stress-induced hyperthermia test, 40 min after drug administration rectal temperature was measured, succinic acid at dose of 1.5 mg/kg, inhibited stress-induced hyperthermia. Thus, these findings indicated that, in contrast with diazepam, succinic acid exhibits anxiolytic-like effect.
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Anxiolytic-like effect of succinic acid in mice
Si Wei Chen*, Qin Xin
, Wei Xi Kong, Li Min, Jing Fang Li
Department of Pharmacology, Shenyang Pharmaceutical University, Box 41, 103 Wenhua Road, 110016 Shenyang, PR China
Received 28 February 2003; accepted 4 June 2003
The putative anxiolytic activity of succinic acid was examined in male mice by using a number of
experimental paradigms of anxiety and compared with that of the known anxiolytic compound diazepam. Use
of the elevated plus-maze test revealed that diazepam (1.0, 2.0 and 4.0 mg/kg, PO) or succinic acid (3.0 or 6.0
mg/kg, PO) increased the percentage of entries into open arms and of time spent on open arms. In novel food
consumption test, succinic acid (3.0, 6.0, and 12.0 mg/kg, IP) caused significant increases in food intake
during 5 min when compared with the vehicle. In the stress-induced hyperthermia test, 40 min after drug
administration rectal temperature was measured, succinic acid at dose of 1.5 mg/kg, inhibited stress-induced
hyperthermia. Thus, these findings indicated that, in contrast with diazepam, succinic acid exhibits anxiolytic-
like effect.
D2003 Elsevier Inc. All rights reserved.
Keywords: Succinic acid; Anxiolytic; Elevated plus-maze; Novel food consumption; Hyperthermia; Mice
Succinic acid is an endogenous metabolite of Krebs cycle and GABA-succinate shunt in the brain
(Sanders et al., 1969). It has been reported that succinic acid had sedative effect (4000 mg/kg) and
protective action against high pressure oxygen convulsion (1180 mg/kg) (Sanders et al., 1969; Gessa
et al., 1968). Jin et al. also reported that succinic acid at dose of 1180 mg/kg produced significant
0024-3205/$ - see front matter D2003 Elsevier Inc. All rights reserved.
* Corresponding author. Tel.: +86-24-23909448; fax: +86-24-24513031.
E-mail address: (S.W. Chen).
Present address: Department of Pharmacology, Jining Medical College, 45 Jianshe Road, 272013 Jining, Shandong
Province, China.
Life Sciences 73 (2003) 3257 3264
decrease of the locomotor activity and hyperthermia and prolonged the pentobarbital sleeping time
significantly in mice. Moreover, succinic acid was found to protect rats against audiogenic seizure
and inhibit the electroshock seizure in mice, but not the convulsion induced by picrotoxin, strychnine
and semicarbazide (Jin and Zhang, 1980). Recently, Yue et al. reported that succinic acid (100400
mg/kg) dose-dependently inhibited pentylenetetrazol chemical and amygdale kindled seizer (Yue et
al., 2002).
The purpose of the present work was to investigate the effects of succinic acid in the elevated plus-
maze, novel food consumption, and stress-induced hyperthermia tests. These animal models have been
validated as tests for anxiety, using behavioral and physiological measures, and have shown good
sensitivity to anxiolytic drugs.
Materials and methods
Male Swiss mice (Experimental Animal center of Shenyang Pharmaceutical University) weighing
2022 g were used. For Elevated plus-maze test, mice were maintained under a 12 h reversed light
cycle (light off 07:00) at room temperature (22 F2jC) for at least 10 day (Ferrari et al., 1998)
For novel food consumption test and stress-induced hyperthermia test, mice were maintained on a
non-reversed 12L: 12D cycle conditions (lights on: 07:00). Five animals were housed in a cage
(25 14 12 cm). Food and water were freely available at all times. Each mouse was used only
Succinic acid was purchased from Shenyang Reagent Co. (Shenyang, China). Diazepam was obtained
from Hubei Pharmaceutical Factory (Hubei, China). Tween 80 was purchased from Shenyang Dongxing
Reagent Factory (Shenyang, China). Succinic acid was dissolved in 0.9% saline, then the solutions were
adjusted to pH 7.2 f7.5 with diluted NaOH. Diazepam was ultrasonically dispersed in distilled water
to which Tween 80 (2 drops/10 ml) had been added. Test drugs were administered PO or IP in a volume
of 10 ml/kg, control animals were administered with the corresponding vehicles. All drugs were freshly
prepared before each treatment trial.
Elevated plus-maze test
The elevated plus-maze comprised two open arms (30 5 cm), facing each other, and two
closed arms (30 515 cm) which extended from a central platform (5 5 cm) to form a
plus sign. The open arms included a clear, slightly raised edge (0.25 cm height) to reduce the
likelihood of animals falling-over. The closed arms were enclosed with three roof-open walls (15
cm height) made from clear plexiglas (Lister, 1987; Cole and Rodgers, 1994). The maze floor was
constructed of black plexiglas. The apparatus was raised to a height of 45 cm by a single central
support (Kuribara et al., 1998). Four 25-W red fluorescent lights arranged as a cross at 100 cm
S.W. Chen et al. / Life Sciences 73 (2003) 3257–32643258
above the maze were used as the source of illumination (Ferrari et al., 1998; Hasenohrl et al.,
Mice were randomly assigned to eight experimental groups and then adjusted slightly to match the
average body weights of the groups (n = 10). All testing was conducted during the midportion of the
dark phase of light cycle and in an order counter balanced for treatment condition and commenced by
placing a mouse on the central platform of the maze facing an open arm. The number of entries into, and
the time spent in, each of the two types of arm, were counted during 5 min. Arm entry was defined as all
four paws having crossed the dividing line between an arm and the central area (Lin et al., 1998). The
maze was thoroughly cleaned with water after each trial to remove any trace of odour. The sessions were
recorded by an overhead video camera linked to a monitor and video recorder in an adjacent laboratory.
All behavioral recordings were carried out with the observer unaware of the treatment of the mice. Test
drugs and the vehicle were administered PO 40 min prior to the plus-maze test.
Novel food consumption in mice
This test was performed according to a procedure described by Keane (Keane et al., 1988). Mice
were subjected to food deprivation for 16 h before the test. Groups of 910 mice were administered
either succinic acid (IP) or diazepam (PO), and 30 min later the animals were placed individually in
plastic cages (25 14 14 cm) for 5 min. Each cage contained a weighed amount of a novel food
source (a mixture of one-third bean oil and two-thirds standard rodent food powder) in a Petri dish. At
the end of the 5 min test period the remaining food and crumb in the cage were collected and weighed
(Britton and Britton, 1981). Food consumption, defined as milligrams eaten in 5 min per gram of body
weight, was calculated for each mouse and the mean ( FSEM) was determined for each treatment
Stress-induced hyperthermia test
The test procedure for stress-induced hyperthermia was similar to that described by Lecci et al. (Lecci
et al., 1990). Temperature measurements were carried out in the same room where mice were housed.
Rectal temperature was measured to the nearest 0.1 jC by a digital thermometer, model JM 624u. The
probe (2-mm diameter), dipped into liquid paraffin before insertion, was held in the rectum until steady
readings were obtained for 20 s and the mouse was reallocated to its cage after each temperature
Eighteen animals were housed per cage (50 25 20 cm). The animals were administered orally
or intraperitoneally with either a drug or vehicle 40 min (succinic acid and its corresponding vehicle) or
50 min (diazepam and its vehicle) before temperature measurement. After diazepam, succinic acid were
given, the rectal temperature was recorded in the first and last three out of 18 mice allocated to each
cage, maintaining the same order as that of treatment, while mice numbers 4 through 15 inclusive were
simply removed and again returned to the cage. The whole procedure required about 10 min. All
experiments were performed between 8 and 12 a.m.
Statistical analyses
Data given represent mean FSEM values. In the elevated plus-maze and novel food consumption
test, data were analyzed by one-way ANOVA. In the stress-induced hyperthermia test, results were
analyzed by factorial analysis of variances (ANOVA). Whenever ANOVA was significant, further
comparisons between vehicle- and drug-treatment groups were performed using the Dunnett’s t-test. All
S.W. Chen et al. / Life Sciences 73 (2003) 3257–3264 3259
analysis was performed using the software SPSS V11.0 for windows. The level of statistical significance
adopted was P< 0.05.
Elevated plus-maze test
In the plus-maze test, conducted 40 min after administration of 3.0, 6.0, and 12.0 mg/kg, succinic acid
did not significantly alter the total number of arm entries made by mice [F(3, 36) = 0.62, P> 0.05, Table
1]. However, succinic acid at doses of 3.0 and 6.0 mg/kg resulted in a significant increase in the
percentage of entries into open arms respect to total arms entries [F(3,36) = 3.92, P< 0.05], as well as
the percentage of time spent in open arms [F(3,36) = 3.20, P< 0.05]. At 12.0 mg/kg, succinic acid-
Table 1
Effects of succinic acid on total arm entries by male mice on the elevated plus-maze
Drug Dose (mg/kg) Total arm entries
Vehicle 15.7 F1.0
Succinic acid 3 18.4 F1.2
6 18.0 F1.3
12 17.9 F2.4
Vehicle 16.3 F1.4
Diazepam 1 31.9 F2.2**
2 24.9 F1.3
4 25.4 F5.4
Values represent means FSEM from 10.
** P< 0.01, Dunnett’s t-test after ANOVA.
Fig. 1. Effects of succinic acid (A) and diazepam (B) in the elevated plus-maze in male mice (n = 10). Results are expressed as
means FSEM. The following parameters are shown: %EOA (percentage of entries into open arms respect to total entries into
the arms); %TOA (percentage of time spent in open arms respect to total time spent in the arms). Mice were given a 5 min test
40 min after PO with diazepam (1, 2, and 4 mg/kg) or succinic acid (3, 6, and 12 mg/kg). Significant differences calculated from
two-tailed Dunnett’ t-test are expressed by *P< 0.05, **P< 0.01 vs. respective vehicle after one way ANOVA.
S.W. Chen et al. / Life Sciences 73 (2003) 3257–32643260
treated mice were not different from vehicle control [Fig. 1(A)]. Diazepam at dose of 1.0 mg/kg
significantly increased the total number of arm entries made by mice [F(3,36) = 4.38, P< 0.01, Table 1].
The ANOVA showed that at doses of 1.0, 2.0, and 4.0 mg/kg, diazepam significantly elevated both the
percentage of entries into open arms and of time spent in open arms [F(3,36) = 3.90, P< 0.05; F(3,36) =
5.08, P< 0.01. Fig. 1(B)].
Novel food consumption test
Fig. 2. shows the mean values of food intake (milligrams per gram) in a 5 min period. Forty minutes
after PO administration, both succinic acid (3, 6, and 12 mg/kg) and diazepam (0.5, and 1 mg/kg)
Fig. 2. Effects of succinic acid and diazepam in the novel food consumption test. Experiment were carried out in groups of
910 mice, each mouse was subjected to food deprivation for 16 h before testing. The values represent the mean FSEM of
food intake (milligrams per gram) in a 5 min period. Significant differences from control: *P< 0.05, **P< 0.01.
Table 2
Effects of administration of succinic acid and diazepam on hyperthermia of mice removed at a later time
Drug Dose Route Time Rectal temperature DT ANOVA interaction
(mg/kg) (min) First three Last three
Vehicle IP 40 37.5 F0.1 38.2 F0.2* 0.7
Succinic acid 1.5 IP 40 37.1 F0.2 37.2 F0.2
0.1 F(3, 64) = 1.68, P> 0.05
3.0 IP 40 37.6 F0.3 37.5 F0.3 0.1
6.0 IP 40 37.0 F0.2 37.6 F0.2* 0.6
Vehicle PO 50 37.4 F0.1 38.3 F0.3** 0.9
Diazepam 1.25 PO 50 37.0 F0.2 37.3 F0.3 0.3 F(3, 64) = 3.31, P= 0.025
2.5 PO 50 36.9 F0.3 37.1 F0.3
5.0 PO 50 37.5 F0.2 36.7 F0.4
Values represent means FSEM from 9 mice. Time indicates the lag time between treatments and test. Dunnett’s t-test.
P< 0.05.
P< 0.01 vs corresponding vehicle; t-test.
*P< 0.05.
**P< 0.01 vs first-three.
S.W. Chen et al. / Life Sciences 73 (2003) 3257–3264 3261
increased the amount of novel food consumption. On the average, the food consumption in the succinic
acid-treated group was significantly different from the vehicle-treated group [F(3,35) = 4.42, P< 0.01].
Diazepam as a reference drug for anxiolytic action, also increased the amount of food consumption [F(3,
35) = 3.27, P< 0.05].
Stress-induced hyperthermia test
The effects of succinic acid at doses of 1.5, 3.0 or 6.0 mg/kg and diazepam at doses of 1.25, 2.5, and
5.0 mg/kg are shown in Table 2. It was observed that the hyperthermia of the last three showed an
significant increase in the vehicle-group. The hyperthermia of the last three in the succinic acid-group
and diazepam-group were not significantly different from the first three except for the succinic acid 6
mg/kg group. Succinic acid (1.5 mg/kg), diazepam (5 mg/kg) inhibited the hyperthermia of the last three
[F(3, 32) = 3.28, p < 0.05; F(3, 32) = 4.65, P < 0.01]. Note that none of the treatments of succinic acid
significantly affected basal core body temperature.
The present study is the first to demonstrate the anxiolytic effect of succinic acid on results from the
elevated plus-maze (a model of anxiety in an approach-avoidance conflict), novel food consumption, and
stress-induced hyperthermia (a model of anticipatory anxiety), which are animal models of anxiety that
have been subjected to through critical appraisal to demonstrate central nervous system activity of drugs.
The elevated plus-maze is an animal analogue that identifies clinically established anxiolytic and
anxiogenic drugs, and is currently used in the screening of drugs that act on the central nervous system
(Hasenohrl et al., 1996; Bhattacharya and Mitra, 1991; Pellow and File, 1986; Pellow et al., 1985).
Normally, mice spent most of their time in the closed arms and avoided the open arms (fear of the open
and high places possibly) (Ang and Hung, 1999). In the test the percentage of entries into open arms and
of time spent in open arms have generally been used as indices of the anxiety. These values were
increased by anxiolytic agents and reduced by anxiogenic agents (Pellow et al., 1985). In this study,
succinic acid at 3 and 6 mg/kg have anxiolytic effects, as indicated by increased the amount of open time
and the number of open entries. At dose of 12 mg/kg, succinic acid did not influence the mice behavior
on the elevated plus-maze. The anxiolytic profile of succinic acid revealed in the present experiment
showed some differences with regard to the known anxiolytic profile of benzodiazepins in this model.
Succinic acid did not affect motor activity. However, diazepam at 1 mg/kg increased motor activity. This
effect of diazepam might be correlated with disinhibition and with ataxia and motor impairment,
Animals generally show neophobia when they are presented with novel food or are placed in a novel
environment (Miyamoto et al., 1992). Anxiolytics facilitate the ‘‘resolution’’ of that apparent neophabia
by increasing the amount of food eaten (Britton and Britton, 1981). In the novel food consumption test,
succinic acid had significantly increased the amount of food intake in mice compared with control. This
finding suggests that succinic acid has reduced fear of eating novel food in mice, and produced
anxiolytic effect.
Various stressors such as handling, restraint, exposure to a novel environment, and foot shock are
known to cause increases in body temperature (Pechnick and Morgan, 1987; York and Regan, 1982).
S.W. Chen et al. / Life Sciences 73 (2003) 3257–32643262
Among animals of the same cage, mice removed last from their home cage always have higher rectal
temperatures than mice removed first from this cage (Lecci et al., 1990; Zethof et al., 1995; Borsini et al.,
1989). This phenomenon is called stress-induced hyperthermia and has been interpreted as being caused
by anticipatory fear of an aversive event (handling). Interestingly, interactions between body temperature
and emotional states seen to exist in man also. The stress-induced rise in rectal temperature can be
blocked by various mechanisms. Benzodiazepine and non-benzodiazepine anxiolytics were capable of
antagonizing stress-induced hyperthermia. The effect of succinic acid on stress-induced hyperthermia
can be considered as specific, because the compound did not affect the core temperature per se:
obviously, succinic acid counteracted the anxiety-dependent variable, and produced anxiolytic activity
similar to diazepam.
Our preliminary study showed that succinic acid did not produce the anxiolytic effect at doses lower
than 1.5 mg/kg or higher than 12 mg/kg such as 24 and 48 mg/kg on the elevated plus-maze
(unpublished data). Succinic acid exhibited an inverted U-shaped dose-response curve for the percentage
of open arm entries based on the present and preliminary set of data. Consistent with many reports
(Pellow and File, 1986; Dawson and Tricklebank 1995), diazepam resulted in a dose-dependent
anxiolytic effect. Succinic acid at 1.5 f12 mg/kg elicited significant anxiolytic effect and did not
cause the change of motor activity. Gessa et al. reported that succinic acid induced a central depressant
action at high doses, for example sedation at 4000 mg/kg (Gessa et al., 1968), this dose are 2666 times
higher than the minimum dose for significant anxiolytic effect (1.5 mg/kg, IP). Thus it is expected that
when the anxiolytic effect is considered as the main effect, the central depressant effect of succinic acid
is very weak.
Succinic acid is widely distributed in the nervous systems. The distribution of succinate in rabbit
revealed that there are regional differences in succinate concentrations within the CNS. Concentrations
of succinate in cerebral cortex, cerebral white matter, and cerebellum are 0.19, 0.24 and 0.35 Amol/g
respectively (Petroff et al., 1988). Succinic acid mainly produces from the tricarboxylic acid cycle and
GABA-succinate shunt in the brain. The possible physiological importance of the GABA-shunt is seen if
one recognizes that succinate markedly stimulates respiration and oxidative phosphorylation. GABA
could serve as a rapidly available source of succinate under physiological stress conditions. It was
reported that succinate oxidation could monopolize the respiratory-electron transport chain which is the
major souce of ATP production. But the effect of low milligram doses of succinic acid on endogenous
levels of this substance in the brain has not been reported.
Although the behavioural effects of succinic acid have been repeatedly reported, the neural
mechanisms underlying these effects have not been clearly elicited. Yue et al. suggested GABAergic
involvement in the anticonvulsion effect of succinic acid (Yue et al., 2002). Given that the role of
receptors in mediation of the anxiolytic effects of benzodiazepines has been well established, it
seems reasonable to propose a GABAergic mechanism for the effects of succinic acid on the results from
the plus-maze, novel food consumption, and stress-induced hyperthermia. Furthermore, we recently
found that the noncompetitive GABA
receptor antagonist, picrotoxin selectively antagonized the
anxiolytic-like effects of succinic acid on the percentage of open arm entries and of time spent in the
open arms of plus-maze anxiety (unpublished data). This suggests that the interaction between succinic
acid and GABA
receptors may be important for the anxiolytic effects of succinic acid.
In conclusion, this study supported the suggestion that succinic acid possesses anxiolytic effects in the
elevated plus-maze, food neophobia and stress-induced hyperthermia models. Furthermore, only low
doses succinic acid were needed to induce an anxiolytic action.
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... The negative ζ-potential prevents the particles from undergoing polyelectrolyte interactions with oppositely charged ions and thus provides the stability and mobility of the conjugates in the vitreous humor environment that consists of polyanionic hyaluronic acid [43]. Succinic anhydride (SA) was chosen as the surface modifier for CS since it is biotransformed in the body into succinic acid, a natural endogenous metabolite of the Krebs cycle, and is therefore non-toxic [44]. Succinylated CS, in addition to its high safety profile and low toxicity, is also less biodegradable than CS, so it is an excellent polymer platform for prolonged glucocorticoid delivery systems and has a release profile for the active pharmaceutical Table 1). ...
... The negative ζ-potential prevents the particles from undergoing polyelectrolyte interactions with oppositely charged ions and thus provides the stability and mobility of the conjugates in the vitreous humor environment that consists of polyanionic hyaluronic acid [43]. Succinic anhydride (SA) was chosen as the surface modifier for CS since it is biotransformed in the body into succinic acid, a natural endogenous metabolite of the Krebs cycle, and is therefore nontoxic [44]. Succinylated CS, in addition to its high safety profile and low toxicity, is also less biodegradable than CS, so it is an excellent polymer platform for prolonged glucocorticoid delivery systems and has a release profile for the active pharmaceutical substance of several months [33,45,46]. ...
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The development of intravitreal glucocorticoid delivery systems is a current global challenge for the treatment of inflammatory diseases of the posterior segment of the eye. The main advantages of these systems are that they can overcome anatomical and physiological ophthalmic barriers and increase local bioavailability while prolonging and controlling drug release over several months to improve the safety and effectiveness of glucocorticoid therapy. One approach to the development of optimal delivery systems for intravitreal injections is the conjugation of low-molecular-weight drugs with natural polymers to prevent their rapid elimination and provide targeted and controlled release. This study focuses on the development of a procedure for a two-step synthesis of dexamethasone (DEX) conjugates based on the natural polysaccharide chitosan (CS). We first used carbodiimide chemistry to conjugate DEX to CS via a succinyl linker, and we then modified the obtained systems with succinic anhydride to impart a negative ζ-potential to the polymer particle surface. The resulting polysaccharide carriers had a degree of substitution with DEX moieties of 2–4%, a DEX content of 50–85 μg/mg, and a degree of succinylation of 64–68%. The size of the obtained particles was 400–1100 nm, and the z-potential was −30 to −33 mV. In vitro release studies at pH 7.4 showed slow hydrolysis of the amide and ester bonds in the synthesized systems, with a total release of 8–10% for both DEX and succinyl dexamethasone (SucDEX) after 1 month. The developed conjugates showed a significant anti-inflammatory effect in TNFα-induced and LPS-induced inflammation models, suppressing CD54 expression in THP-1 cells by 2- and 4-fold, respectively. Thus, these novel succinyl chitosan-dexamethasone (SucCS-DEX) conjugates are promising ophthalmic carriers for intravitreal delivery.
... The functional properties of PGSu are similar to those of PGS. Succinic acid is a natural metabolite in the Krebs cycle; therefore, it is non-toxic to mammalian organisms [34]. The FDA (Food and Drug Administration)-approved succinic acid as a component of medical and pharmaceutical devices [35]. ...
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Glycerol polyesters have recently become objects of interest in tissue engineering. Barely known so far is poly(glycerol itaconate) (PGItc), a biocompatible, biodegradable polyester. Due to the presence of a C=C electron-acceptor moiety, it is possible to post-modify the product by Michael additions to change the properties of PGItc. Thus, using PGItc as one of the elements of cellular scaffold crosslinked in situ for bone tissue regeneration seems to be a very attractive yet unexplored solution. This work aims to optimize the synthesis of PGItc to obtain derivatives with a double bond in the side chain with the highest conversion rates. The experiments were performed with itaconic anhydride and glycerol using mathematical planning of experiments according to the Box-Behnken plan without solvent and catalyst. The input variables of the process were the ratio of the OH/COOH, temperature, and reaction time. The optimised output variables were: the degree of esterification (EDtitr), the degree of esterification calculated from the analysis of 1H NMR spectra (EDNMR), and the degree of itaconic anhydride conversion—calculation based on 13C NMR spectra (%X13CNMR). In each of statistical models, the significance of the changed synthesis parameters was determined. Optimal conditions are when OH/COOH ratio is equal to 1.5, temperature is 140 °C and time of reaction is 5 h. The higher OH/COOH ratio, temperature and longer the experiment time, the higher the value of the degree of esterification and the degree of anhydride conversion.
... To the best of our knowledge, no reports have identified altered succinic acid levels in humans with PTSD. Succinic acid has anxiolytic-like effects in rodents and it is possible that the lower levels of succinic acid in individuals with PTSD may promote anxiety (75). Another metabolite that was significantly different between veterans with PTSD and controls was acetic acid. ...
Introduction: Posttraumatic stress disorder (PTSD) is a psychiatric disorder, resulting from exposure to traumatic events. Current recommended first-line interventions for the treatment of PTSD include evidence-based psychotherapies, such as Cognitive Processing Therapy (CPT). Psychotherapies are effective for reducing PTSD symptoms, but approximately two-thirds of veterans continue to meet diagnostic criteria for PTSD after treatment suggesting there is an incomplete understanding of what factors sustain PTSD. The intestine can influence the brain and this study evaluated intestinal readouts in subjects with PTSD. Methods: Serum samples from controls without PTSD (n=40) from the Duke In-Trust Program were compared to veterans with PTSD (n=40) recruited from the Road Home Program at Rush University Medical Center. Assessments included microbial metabolites, intestinal barrier, and intestinal epithelial cell function. Additionally, intestinal readouts were assessed in PTSD subjects before and after a 3-week CPT-based intensive treatment program (ITP) to understand if treatment impacts the intestine. Results: Compared to controls, veterans with PTSD had a pro-inflammatory intestinal environment including lower levels of microbiota-derived metabolites acetic, lactic, and succinic acid, intestinal barrier dysfunction (lipopolysaccharide (LPS) and LPS binding protein), an increase in HMGB1, and a concurrent increase in the number of intestinal epithelial cell-derived extracellular vesicles. The ITP improved PTSD symptoms but no changes in intestinal outcomes were noted. Conclusion: This study confirms the intestine is abnormal in subjects with PTSD and suggests that effective treatment of PTSD does not alter intestinal readouts. Targeting beneficial changes in the intestine may be an approach to enhance existing PTSD treatments.
... barrier to boost cerebellar function, thus to improve motor activity, suggesting a direct action on the brain (34). Also, oral administration of succinate can increase the amount of food intake in mice (35). A previous study using another organic acid, acetate, has shown a parasympathetic nerve-dependent stimulation mechanism, whereby both acetate feeding and intracerebroventricular injection can promote fish feed intake and upregulate orexigenic genes in the brain (15). ...
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Succinate is widely used in the food and feed industry as an acidulant, flavoring additive, and antimicrobial agent. This study investigated the effects of dietary succinate on growth, energy budget, nutritional metabolism, protein succinylation, and gut microbiota composition of zebrafish. Zebrafish were fed a control-check (0% succinate) or four succinate-supplemented diets (0.05, 0.10, 0.15, and 0.2%) for 4 weeks. The results showed that dietary succinate at the 0.15% additive amount (S0.15) can optimally promote weight gain and feed intake. Whole body protein, fat, and energy deposition increased in the S0.15 group. Fasting plasma glucose level decreased in fish fed the S0.15 diet, along with improved glucose tolerance. Lipid synthesis in the intestine, liver, and muscle increased with S0.15 feeding. Diet with 0.15% succinate inhibited intestinal gluconeogenesis but promoted hepatic gluconeogenesis. Glycogen synthesis increased in the liver and muscle of S0.15-fed fish. Glycolysis was increased in the muscle of S0.15-fed fish. In addition, 0.15% succinate-supplemented diet inhibited protein degradation in the intestine, liver, and muscle. Interestingly, different protein succinylation patterns in the intestine and liver were observed in fish fed the S0.15 diet. Intestinal proteins with increased succinylation levels were enriched in the tricarboxylic acid cycle while proteins with decreased succinylation levels were enriched in pathways related to fatty acid and amino acid degradation. Hepatic proteins with increased succinylation levels were enriched in oxidative phosphorylation while proteins with decreased succinylation levels were enriched in the processes of protein processing and transport in the endoplasmic reticulum. Finally, fish fed the S0.15 diet had a higher abundance of Proteobacteria but a lower abundance of Fusobacteria and Cetobacterium. In conclusion, dietary succinate could promote growth and feed intake, promote lipid anabolism, improve glucose homeostasis, and spare protein. The effects of succinate on nutritional metabolism are associated with alterations in the levels of metabolic intermediates, transcriptional regulation, and protein succinylation levels. However, hepatic fat accumulation and gut microbiota dysbiosis induced by dietary succinate suggest potential risks of succinate application as a feed additive for fish. This study would be beneficial in understanding the application of succinate as an aquatic feed additive.
... 18 The situation is similar in succinic acid, which is (as ionized form) an essential metabolite of the Krebs cycle in the mitochondrial matrix. 19 Succinic acid is quite common in nature in most fruits and vegetables. 20 Adipic acid is not, however, one of the essential compounds in terms of metabolic processes occurring in living organisms. ...
Nowadays, glycerol polyesters are willingly studied, due to their potential to use in tissue engineering and drug delivery systems. Poly(glycerol citrate) is a quite new material that could have very interesting properties because of the easy formation of three-dimensional hydrophilic networks. Moreover, both glycerol and citric acid are commonly used chemicals in the food and pharmaceutical industry. They are cheap, well available, and well-known substances. In this work poly(glycerol citrate) was synthesized in bulk polycondensation with water removal. The polyester structure was widely characterized by FTIR, NMR, GPC, and the end group titration methods. The synthesis procedure was optimized and described via five mathematical models with the use of the design of experiments. The presented method was developed for further industrial use and allows for avoiding gelling phenomena. Based on this work, poly(glycerol citrate) could be obtained with desired properties, moreover, the process is prepared for scaling-up.
... Camphore, camphene, borneol, terpinene-4-ol, p-cymene, fenchol, isoborneol, abietic acid, pimaric acid and isopimaric acid inhibit the biological and biochemical functions of organisms, as well as inhibiting the growth of bacteria (P. aeruginosa, Escherichia coli, S. aureus and K. pneumoniae) [31]. Also, these compounds available in the amber make it potentially useful for the preparation of modern medicines because they exhibit antibacterial, antifungal, anti-inflammatory, insecticidal and antioxidant potential, P-cymene, limonene, terpinen-4-ol and pulegone exhibit antioxidative properties. ...
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Amber is a fossil residue from an etched resin from the ancient, used as a medicinal agent to control microbial infections, due to containing chemical compounds that detected by GC-MS analysis. Antibacterial activity was detected for both Dimethyl sulfoxide and ethanolic extracts of orange and brown amber. The largest zones of inhibition were 19 mm and 16 mm for Dimethyl sulfoxide extract of orange and brown amber at 250 mg/ml on Staphylococcus aureus and Pseudomonas aeruginosa respectively, while the largest inhibition zones of the ethanolic extracts were 20 and 22 mm for orange and brown amber at 100 mg/ml on E. coli. The GC-MS analysis revealed a total of 35 compounds in Baltic amber. Major chemical components identified in orange and brown amber included borneol (16.80% and 17.60%), isopimar acid methyl aster (17% and 13.65%), camphor (8.15% and 7.04%), 2-Fenchanol (7.44% and 7.76%), and m-cymene (6.24% and 5.40 %), respectively. Orange amber contains seven monoterpenes, six sesquiterpenes, and three diterpenes, while brown amber contains seven monoterpenes, four sesquiterpenes, and one diterpene.
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The anxiolytic effect of Eurycoma longifolia Jack in mice was examined. Fractions of E. longifolia Jack extract produced a significant increase in the number of squares crossed (controls=118.2±10.2 squares), but significantly decreased both the immobility (controls=39.4±4.0 sec) and fecal pellets (controls=12.3±2.1 fecal pellets) when compared with control mice in the open-field test; they significantly increased the number of entries (controls=6.7±0.5 entries) and time spent (controls=42.9±0.1 sec) in the open arms, but decreased both the number of entries (controls=13.2±0.7 entries) and time spent (controls=193.4±0.7 sec) when compared with the control mice in the closed arms of the elevated plusmaze test. Furthermore, fractions of E. longifolia Jack extract decreased the fighting episodes significantly (controls=18.0±0.4 fighting episodes) when compared with control mice. In addition, these results were found to be consistent with anxiolytic effect produced by diazepam. Hence, this study supports the medicinal use of this plant for anxiety therapy.
Age-related behavioral changes in the passive avoidance, food neophobia, elevated plus-maze, and water-lick conflict tests were studied using substrains of senescence-accelerated mouse (SAM-P/8 and SAM-R/1) at 2 to 20 months of age. SAM-P/8 mice exhibited a significant impairment of acquisition of passive avoidance compared with SAM-R/1 mice when they were trained repeatedly, and the acquired response in SAM-P/8 mice rapidly diminished in contrast to good retention in SAM-R/1 mice. SAM-P/8 mice showed an age-related decrease in the latency to eat novel food after a 24-h food deprivation as compared with SAM-R/1 mice at 2 to 12 months of age, despite no significant difference in latency to eat familiar food between the two strains. In the elevated plus-maze test, SAM-P/8 mice had apparent increases in the number of entries into open arms and time spent on open arms in comparison to SAM-R/1 mice at 4 through 12 months of age; this difference became obvious with aging, implying age-associated reduced anxiety in the SAM-P/8 strain. In addition, SAM-P/8 mice exhibited a significant increase in punished water drinking compared to SAM-R/1 mice in the water-lick conflict test, although unpunished water intake in SAM-P/u mice did not differ from that in the SAM-R/1 control. Aged SAM-R/1 mice, 20 months old, exhibited low anxiety-like behavior in the food neophobia and elevated plusmaze tests such as was seen in SAM-P/8 mice, when compared with young (4-month-old) SAM-R/1 mice. These results indicate that the SAM-P/8 strain shows age-related reduced anxiety-like behavior compared to the SAM-R/1 strain, which may be related to learning and memory impairment in the strain. It is suggested that the SAM-P/8 strain may be a useful animal model for studying emotional disorders and related memory impairments in dementia, which can be used at an early stage of age.
Propylene glycol is a common solvent often contained in injectable solutions of anxiolytics of low water-solubility, such as diazepam (Valium) and pentobarbital (Nembutal). Several studies have shown that propylene glycol can have an inhibitory effect on the central nervous system. This study, using ethanol for comparison, further examined whether propylene glycol has anti-anxiety properties. Use of the elevated plus-maze test with male mice revealed that propylene glycol at doses (27 or 41 mmol kg−1, i.p.) which did not affect general activity, increased the number of entries into open arms and of head dips over open arm edges, indicative of an anxiolytic effect. In parallel, ethanol (14 and 27 mmol kg−1, i.p.) caused an increase in the amount of time spent on open arms and number of entries into open arms, accompanied by reduction of returns into closed arms. These doses of ethanol had no significant effect on motor ability. The results suggest that propylene glycol can act as an anxiolytic agent and that its anxiolytic potency is weaker than that of ethanol. In addition to previous warnings about the pharmacological effects of propylene glycol, the findings of this study alert investigators to the anxiolytic properties of the compound when it is employed as a solvent in anxiety or anxiety-related studies.
To investigate whether an elevated plus-maze consisting of two open and two closed arms could be used as a model of anxiety in the mouse, NIH Swiss mice were tested in the apparatus immediately after a holeboard test. Factor analysis of data from undrugged animals tested in the holeboard and plus-maze yielded three orthogonal factors interpreted as assessing anxiety, directed exploration and locomotion. Anxiolytic drugs (chlordiazepoxide, sodium pentobarbital and ethanol) increased the proportion of time spent on the open arms, and anxiogenic drugs (FG 7142, caffeine and picrotoxin) reduced this measure. Amphetamine and imipramine failed to alter the indices of anxiety. The anxiolytic effect of chlordiazepoxide was reduced in mice that had previously experienced the plus-maze in an undrugged state. Testing animals in the holeboard immediately before the plus-maze test significantly elevated both the percentage of time spent on the open arms and the total number of arm entries, but did not affect the behavioral response to chlordiazepoxide. The plus-maze appears to be a useful test with which to investigate both anxiolytic and anxiogenic agents.
A novel test for the selective identification of anxiolytic and anxiogenic drug effects in the rat is described, using an elevated + -maze consisting of two open arms and two enclosed arms. The use of this test for detecting such drug effects was validated behaviourally, physiologically, and pharmacologically. Rats made significantly fewer entries into the open arms than into the closed arms, and spent significantly less time in open arms. Confinement to the open arms was associated with the observation of significantly more anxiety-related behaviours, and of significantly greater plasma corticosterone concentrations, than confinement to the closed arms. Neither novelty nor illumination was a significant contributor to the behaviour of the rats on the + -maze. A significant increase in the percentage of time spent on the open arms and the number of entries into the open arms was observed only within clinically effective anxiolytics (chlordiazepoxide, diazepam and, less effectively, phenobarbitone). Compounds that cause anxiety in man significantly reduced the percentage of entries into, and time spent on, the open arms (yohimbine, pentylenetetrazole, caffeine, amphetamine). Neither antidepressants nor major tranquilisers had a specific effect. Exposure to a holeboard immediately before placement on the + -maze showed that behaviour on the maze was not clearly correlated either with exploratory head-dipping or spontaneous locomotor activity.
In group-housed mice (ten per cage), mice removed last from their home cage always have higher rectal temperatures than mice removed first from this cage. Stress-induced hyperthermia is calculated as the difference (ΔT) between the basal temperature (mouse number 1) and the end temperature (mouse number 10) when the temperature of the ten mice is sequentially measured using a 1-min interval between rectal measurements. Using this protocol, various drugs, belonging to different pharmacological classes, were tested in order to investigate their putative anxiolytic effect, measured as a decrease in ΔT. Benzodiazepines (diazepam, alprazolam), alcohol, and some (flesinoxan, buspirone), but not all (ipsapirone) 5-HT1A receptor agonists had anxiolytic properties with this protocol. Clonidine (α2-adrenoceptor agonist) and prazosine (α1-adrenoceptor antagonist) had, but at high doses, some anxiolytic actions. Antidepressants (desipramine, fluvoxamine, nomifensine, tianeptine. amitriptyline, clomipramine, imipramine), serotonergic ligands (ondansetron, ketanserin, 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI), fenfluramine, metachlorophenylpiperazine (mCPP), eltoprazine) and various other drugs (phenobarbital, pentetrazol, haloperidol, apomorphine, amphetamine, (+)-N-[1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-1,4-benzodiazepin-3(R)-yl]-N′-(3-methylphenyl)urea (MSD 365260), dizocilpine and acetyl salicylic acid) had no anxiolytic activity. The stress-induced hyperthermia protocol used was unable to detect anxiogenic properties of drugs, probably due to a (physiological) ceiling in the maximal end temperature. The stress-induced hyperthermia protocol with mice can be used to measure anxiolytic properties of drugs and is a fast and robust model which does not need extensive training of animals.
Anxiolytic drugs including diazepam (DZP), chlordiazepoxide (CDX), pentabarbitol (PB) and ethanol (EtOH) produce specific alterations in the behavior of fasted rats given access to a single food pellet secured in the center of a novel open field environment. These drugs increase the total amount of food eaten in a 15 mind test and the mean amount eaten per approach to the fast food pedestal. This latter appears to be the more sensitive index of anxiolytic drug action and occurs at doses which have no effect on rearing or grooming. DZP was effective following either acute or chronic (15 day) treatment at doses which had no effect on the food consumption by fasted rats tested in their home cages. The effects of the sedative benzodiazepine, flurazepam, were similar to those of DZP but were not statistically significant. Behavioral effects similar to those of DZP were seen in animals received additional handling prior to testing or in animals habituated to the open field. Neither the anti-psychotic haloperidol nor morphine mimicked the actions of DZP.
The putative anxiolytic activity of the white and red varieties of ginseng, the root of Panax ginseng, was investigated in rats and mice using a number of experimental paradigms of anxiety and compared with that of diazepam. Pilot studies indicated that single-dose administration of ginseng had little to no acute behavioural effects, hence the two varieties of ginseng were administered orally at two dose levels twice daily for 5 days, while diazepam (1 mg/kg, i.p.) was administered acutely. White and red varieties of ginseng (20 and 50 mg/kg) showed positive results when tested against several paradigms of experimental anxiety. Both were effective in the open-field and elevated plus-maze tests and reduced conflict behaviour in thirsty rats and footshock-induced fighting in paired mice. Ginseng also attenuated pentylenetetrazole-induced decrease in rat brain MAO activity, confirming its anxiolytic activity since this has been proposed to be an endogenous marker for anxiety. The effects induced by white and red ginseng (50 mg/kg x 5 days) were comparable to those induced by diazepam (1 mg/kg).
The current study investigates the action of anxiolytics, antidepressants, neuroleptics, antipyretics, muscle relaxants, antihypertensives and naloxone in a novel animal model of anxiety, based on the evidence that mice removed last from their cage develop hyperthermia (stress-induced hyperthermia, SIH) when compared to those removed first. Alprazolam (0.15-0.6 mg/kg), chlordiazepoxide (25 mg/kg), estazolam (1 mg/kg), phenobarbital (20 mg/kg), ethanol (2 and 4 g/kg), buspirone (5 and 10 mg/kg) and prazosin (1 and 2 mg/kg), as well as repeatedly administered diazepam (5 mg/kg), inhibited SIH. In contrast, tofisopam (12.5-200 mg/kg), desipramine (15 and 30 mg/kg), amitriptyline (10 mg/kg), fluoxetine (10 and 20 mg/kg), tranylcypromine (5 and 10 mg/kg), chlorpromazine (1 and 2 mg/kg), clozapine (2 and 4 mg/kg), pimozide (0.5 and 1 mg/kg), l-sulpiride (15 and 30 mg/kg), l-propranolol (5 and 10 mg/kg), acetyl salicylic acid (200 and 400 mg/kg), indomethacin (2.5 and 5 mg/kg), verapamil (2.5 and 5 mg/kg), captopril (25 and 50 mg/kg), dantrolene (10 and 20 mg/kg), mephenesin (300 and 600 mg/kg), d-amphetamine (1 and 4 mg/kg) and naloxone (2.5 and 15 mg/kg) were inactive, as were 10 mg/kg imipramine, amitriptyline and fluoxetine injected every day for 21 days. Reserpine at high doses (1.25 and 2.5 mg/kg) but not at a lower dose (0.62 mg/kg) prevented SIH, but in this case animals showed a behavioural syndrome which could have interfered with the occurrence of the hyperthermia.