Anti-oxidation and lifespan-extension activities of ReishiMax in oxidative stress and aging models

Abstract

Background: Ganoderma lucidum (Reishi) has long been used as a medicinal herb for immune modulation, cancer prevention and adjuvant therapy. ReishiMax, a proprietary extract of G. lucidum (6% Reishi triterpenes, 13.5% Reishi polysaccharides), enhances proliferations of macrophages, B, T and NK lymphocytes, increases serum IgA, IgG, IgM & secretion of IL2 and Interferon, decreases IL5 secretion, and inhibits cancer malignancy in intro and in mice models.
Objective: This study was to examine the anti-oxidation and lifespan-extension effects of ReishiMax in oxidative-stress and normal aging mouse models.
Methods: In the anti-oxidation study, ICR mice were randomized to receive vehicle or ReishiMax at a dose of 175, 350 or 700 mg/kg for 60 days via gavage. They received a single dose of 11 Gy 60Co γ-irradiation on Day 60 and sacrificed on Day 64. Anti-oxidation markers were tested in plasma or liver homogenates, including catalase (CAT), superoxide dismutase (SOD), plasma total thiol groups, glutathione peroxidase (GSH-Px), glutathione reductase (GSH-Rd). In the lifespan study, ICR mice (12 months of age; 100 males & 100 females) were randomized to receive either rat chaw alone or the forage prior mixed with ReishiMax at a dose of 175, 350 or 700 mg/kg. Calorie intake was adjusted twice a week to match the control levels for males and females. Death in each groups was recorded every day until all mice die, followed by computing average lifespan, maximal lifespan (average of longest 10% lifespan), and survival plot.
Results: In the anti-oxidation study, γ-irradiation reduced plasma total thiol groups, RBC CAT & SOD, liver CAT, GSH-Px, GSH-Rd, compared to non-irradiation controls. Compared to irradiation controls, 60 days of oral ReishiMax increased RBC SOD by 9-17% (p=0.027; 0.037; p<0.001), RBC CAT by 55-64% and liver CAT by 19-30% (all p<0.01). Oral ReishiMax at a dose of 350 mg/kg increased the plasma thiol groups by 12% (p=0.008). ReishiMax therapy (175 or 350mg/kg) increased liver GSH-Rd by 8.8-9.2% (p=0.03). ReishiMax therapy (700mg/kg) increased liver GSH-Px by 10% (p=0.003). In the lifespan study, ReishiMax treatment showed: ①no significant differences in body weight and calorie intake among the groups; ②compared to controls, the lifespan was extended 30-66 days at 50% survival, 46-110 days at 20% survival, 46-121 days at 4% survival, and 186-262 days (19.5%-27.5%) at 0% survival (the longest lifespan individuals). Kaplan-Meier survivor analysis showed significant extensions of lifespan and reduction of death risks with ReishiMax, with the best survivor curve for ReishiMax 175 mg/kg therapy (equivalent to the human dose) (p=0.023). ③The average lifespan of control mice was 665±151 days, with no significant difference between genders. Compare to controls, the average lifespan of mice in the 3 ReishiMax dosage groups was extended 63 days (9.5%), 68days (10.3%), and 45 days (6.8%), respectively. ④The maximal lifespan of control mice was 910±44 days, with no significant difference between genders. Compare to controls, the maximal lifespan of mice in the 3 ReishiMax dosage groups was significantly extended 191 days (21.0%), 134 days (14.8%), and 124 days (13.7%), respectively.
Conclusion: Our data demonstrated that ReishiMax enhances body’ anti-oxidant function as one of the potential anti-aging mechanisms, and extends the lifespan in normal aging mice. Using ReishiMax as the example, our study proves the anti-aging belief of G. lucidum, the belief has been held in Chinese society for thousands of years.

Full text

Anti-oxidation and lifespan-
extension activities of ReishiMax
in oxidative stress and aging
models
灵芝提取物ReishiMax抗氧化及延长
小鼠寿命的实验研究
Jieying Yang
Pharmanex Beijing Clinical Pharmacology Center

实验一：动物抗氧化实验
Animal anti-oxidation study
60Co-γ射线照射诱导小鼠氧化压力模型
Oxidative damage aging mice
实验二：动物生存实验
Animal life span study
自然衰老小鼠
Natural aging mice
灵芝提取物（RM）抗氧化及延长小鼠寿命的研究
ReishiMax Anti-aging studies in animal models

实验一：灵芝提取物（RM）在氧化压力小鼠中抗氧化作用
To Examine the Anti-oxidant Effect of ReishiMax in
Oxidative Stress Male Mice
0 (days) 60 64
Collect: Liver Blood
11 Gy of 60Co
Non-irradiation control (n=15)
ReishiMax 700 mg/kg (n=15)
Irradiation control (n=15)
ReishiMax 175 mg/kg (n=15)
ReishiMax 350 mg/kg (n=15)

灵芝提取物增加血浆中巯基含量
Increases in Plasma Thiol Groups in Mice Given
ReishiMax and 60Co Irradiation
0.4
0.5
0.6
0.7
0.8
0.9
1.0
+ 25.6%
P<0.001
+ 12.8%
P=0.039
+ 14.3%
P=0.016
+ 21.9%
P<0.001
Plasma thiol groups(mmol/L)
Non-
irradiation
control
Irradiation
control
175 350 700
ReishiMax (mg/kg)
60Co -11Gy

RBC SOD (U/g Hb)
Non-
irradiation
control
Irradiation
control
175 350 700
ReishiMax (mg/kg)
60Co -11Gy
6,000
7,000
8,000
9,000
10,000
11,000
12,000
+ 11.2%
P<0.001
+ 10.1%
P=0.005
+ 9.5%
P=0.009
+ 17.8%
P<0.001
灵芝提取物增加红细胞中SOD活性
Increases in RBC SOD in Mice Given
ReishiMax and 60Co Irradiation

灵芝提取物增加肝脏及红细胞中CAT活性
Increases in Liver & RBC CAT in Mice Given
ReishiMax and 60Co Irradiation
0
3
6
9
12
15
18
+ 42.4%
P<0.001
+ 55.0%
P<0.001
+ 60.6%
P<0.001
+ 64.3%
P<0.001
Non-
irradiation
control
Irradiation
control
175 350 700
ReishiMax (mg/kg)
60Co -11Gy
RBC
(U/mg protein)
Non-
irradiation
control
Irradiation
control
175 350 700
ReishiMax (mg/kg)
60Co -11Gy
0
5
10
15
20
25
30
35
40
+ 38.1%
P<0.001
+ 30.1%
P<0.001
+ 19.2%
P0.005
+ 23.1%
P=0.001
Liver
(U/mg protein)

灵芝提取物增加肝脏GSH-Px活性
Increases in Liver GSH-Px in Mice Given
ReishiMax and 60Co Irradiation
GSH-Px (U/mg protein)
Non-
irradiation
control
Irradiation
control
175 350 700
ReishiMax (mg/kg)
60Co -11Gy
200
250
300
350
400
450
500
+ 15.0%
P<0.001
+ 10.5%
P=0.003

灵芝提取物增加肝脏GSH-Px活性
Increases in Liver GSH-Px in Mice Given
ReishiMax and 60Co Radiation
GSH-Rd (U/mg protein)
Non-
irradiation
control
Irradiation
control
500 1000 1500
ReishiMax (mg/kg)
60Co -11Gy
0
4
8
12
16
20
24
+ 14.6%
P<0.001
+ 8.8%
P=0.034
+ 9.2%
P=0.026

实验一小结 Summary (1)
经灵芝提取物治疗后：
ReishiMax treatment causes:
↑血浆中巯基含量
Plasma thiol groups
↑红细胞中SOD活性
RBC SOD
↑肝脏和红细胞中CAT活性
Liver & RBC CAT
↑肝脏中谷胱甘肽过氧化物酶和谷胱甘肽还
原酶活性
↑Liver GSH-Px and GSH-Rd

实验二：灵芝提取物（RM）在自然衰老小鼠中
延长生存时间的作用
To Examine the Lifespan Extension Effect of
ReishiMax in Natural Aging Mice
Placebo control (n=50)
RM 700 mg/kg (n=50)
RM 350 mg/kg (n=50)
RM 175 mg/kg (n=50)
~ 3 years
All Mice Death
12 m-old

Time (weeks)
Body Weight (g)
20
25
30
35
40
45
50
55
010 20 30 40 50 60 70
RM 175 RM 700
Control
RM 350
雄性小鼠体重变化情况
Body Weight Changes in Males

雌性小鼠体重变化情况
Body Weight Changes in Females
Time (weeks)
Body Weight (g)
20
25
30
35
40
45
50
55
010 20 30 40 50 60
RM 175 RM 700
Control
RM 350

Time (days)
灵芝提取物延长小鼠的生存时间
ReishiMax Extended Lifespan
Kaplan-Meier Cumulative Survival Curves
Cumulative Survival Rate
365 565 765 965 1165 1365
0
0.2
0.4
0.6
0.8
1.0
Control
RM 700
RM 350
RM 175
灵芝提取物延长小鼠的生存时间
ReishiMax Extended Lifespan

灵芝提取物延长小鼠的生存时间
ReishiMax Extended Lifespan
Cumulative Survival Rate
Time (days)
Kaplan-Meier Cumulative Survival Curves
0
0.2
0.4
0.6
0.8
1.0
365 565 765 965 1165 1365
Control
RM 175
P=0.0226

灵芝提取物延长小鼠的生存时间
ReishiMax Extended Lifespan
Cumulative Survival Rate
Time (days)
Kaplan-Meier Cumulative Survival Curves
365 565 765 965 1165
0
0.2
0.4
0.6
0.8
1.0
Control
RM 350
P=0.0281

030 60 90 120 150 180 210 240 270
90%
70%
50%
30%
20%
10%
4%
0%
lifespan extended (days)
% survival
90% 70% 50% 30% 20% 10% 4% 0%
700mg/kg 44 830 45 46 61 46 242
350mg/kg 56 49 65 51 53 98 121 186
175 mg/kg 20 13 66 50 110 182 120 262
不同存活率，灵芝提取物延长小鼠的生存时间
Lifespan Extended (Days) at the % Survival

灵芝提取物延长小鼠平均寿命
ReishiMax Extended Average Lifespan
600
620
640
660
680
700
720
740
760
780
Placebo
control
175 350 700
Average lifespan (days)
ReishiMax (mg/kg.bw)
+ 69 days
+ 10.3%
+ 64 days
+ 9.6% + 45 days
+ 6.8%

The longest 10% lifespan (days)
Placebo
control
ReishiMax
500 mg/kg
ReishiMax
1500 mg/kg
ReishiMax
1000 mg/kg
P=0.003
P=0.049
P=0.033
800 900 1000 1100 1200 1300
灵芝提取物延长小鼠最高寿命
ReishiMax Extended maximal Lifespan
+191days
+134days
+124days

实验二小结 Summary (2)
灵芝提取物对小鼠寿命的影响：
ReishiMax extended mice lifespan:
↑Kaplan-Meier生存分析显示，RM延长小鼠生存时
间，降低死亡风险
Kaplan-Meier survivor analysis showed significant
extensions of lifespan and reduction of death risks
↑治疗组小鼠死亡中位数分别延长64,69和35天
The median lifespan of mice is extended for 64，69，35 days
↑三个治疗组小鼠平均寿命延长45-69天
The median lifespan of mice is increased 10-66 days
↑小鼠的最高寿命显著延长
The maximal lifespan of mice is significantly extend

总结 Conclusions
灵芝提取物（ReishiMax）：
实验一：动物抗氧化实验
Animal anti-oxidation study
有效减少体内氧化损伤，改善机体抗氧化能力
Reduce the body’ oxidative damage and
enhances anti-oxidant function
实验二：动物生存实验
Animal life span study
延长正常老龄小鼠的寿命
Extends the lifespan in normal aging mice