Evaluating pulmonary toxicity of Shuang-Huang-Lian in vitro and in vivo.
ABSTRACT ETHNOPHARMAOCOLOGICAL RELEVANCE: Shuang-Huang-Lian (SHL) is a traditional Chinese formula and has been used for the treatment of respiratory tract infections by inhalation. However, the pulmonary toxicity via inhalation is largely uninvestigated.
To evaluate the pulmonary toxicity of SHL following in vivo intratracheal spray to rats and in vitro exposures to A549 and Calu-3 cells.
Calu-3 and A549 cells were exposed to SHL, chlorogenic acid, baicalin and forsythin solutions and in vitro cytotoxicity was evaluated using an MTT assay, whilst rats were subjected to intratracheal administration of SHL solutions and in vivo toxicity was indicated by assaying the LDH activity and total protein content in bronchoalveolar lavage fluid (BALF) and observing the histopathologic changes of the lungs. Secretion of inflammatory mediators, including IL-6, IL-8 and TNF-α, in cell culture media and BALF was quantified by ELISA.
The MTT cell viability data revealed the presence of minor toxicity to Calu-3 or A549 cells following exposure to SHL and its major ingredients for 24h or 48 h. However, the cell cultural media showed no sign of inflammatory responses. The in vivo results showed that exposures to SHL at doses of up to 50mg/kg did not significantly increase the total protein content, the LDH activity and the concentrations of IL-6, IL-8 and TNF-α in BALF. However, although intratracheal sprayed SHL at doses of up to 6 mg/kg for histopathologic study and up to 25mg/kg for cell counts showed no sign of adverse effects, inhaled SHL at elevated doses appeared to induce alveolar fusion in the lung and significant increases in the cell number of monocytes and granulocytes in the BALF.
The results demonstrated that the pulmonary safety of inhaled SHL was dependent on the administered dose. Inhalation therapy of SHL may be safely used when the inhaled dose was properly controlled.
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ABSTRACT: The development of dry powder inhalation (DPI) products of traditional Chinese medicine (TCM) remains to be a challenge due to chemical complexity and batch-to-batch variations in constituent composition. This study was to investigate the feasibility of using spray-dried corrugated particles to improve the aerodynamic performance of a TCM, Shuang-Huang-Lian (SHL), in carrier-based DPI. Particles with different surface roughness were spray-dried by the addition of leucine and concomitant manipulation of spray-drying parameters. The surface roughness was determined by atomic force microscopy, whilst the aerodynamic performance of drug particle-mannitol/lactose blends was evaluated using a next-generation pharmaceutical impactor through a Cyclohaler. Although the emission efficiency for corrugated particle-based DPI was ~10% lower than that for smooth SHL, the fine particle fractions (FPF(<4.4 μm)) of 32.4-36.8% for the former were significantly higher than those of 14.7-16.2% for the latter. In particular, the FPF and fraction of drug detached from the carrier appeared not to be significantly affected by the variation in constituent composition of SHL. This study demonstrates that the use of corrugated particles in carrier-based DPI improved aerosol performance by facilitating drug detachment from the carrier, independent of variation in constituent composition, and such particles were potentially applicable to the development of SHL DPI products.AAPS PharmSciTech 05/2012; 13(3):816-25. · 1.58 Impact Factor
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ABSTRACT: A sensitive and specific high performance liquid chromatography coupled with mass spectrometric (LC-MS) method was developed and validated for the simultaneous determination of three main active constituents of Shuang-huang-lian injection with and without the combination use of levofloxacin injection in rat plasma. After addition of the internal standard rutin, plasma samples were protein precipitated with acetonitrile, the chromatographic separation was achieved on a Kromasil C₁₈ column (250 mm × 4.6 mm, 5 μm), using a gradient mobile phase system of acetonitrile-water containing 0.05% formic acid. The analytes were detected without interference in the selected ion monitoring (SIM) mode with positive electrospray ionization. The linear range was 0.04-20 μg/mL for chlorogenic acid, 0.8-400 μg/mL for baicalin and 0.01-5.0 μg/mL for phillyrin, respectively. The accuracy (relative error, R.E.%) were between -2.7 and 3.4%, while the intra-day and inter-day precisions were less than 9.2 and 9.6% for the three analytes, respectively. This method was successfully applied to the drug interaction study of Shuang-huang-lian freeze-dried powder combined with levofloxacin injection after intravenous administration to rats. The results indicated that there were obvious differences in the pharmacokinetic behaviors after combination compared with only administration of Shuang-huang-lian injection.Journal of chromatography. B, Analytical technologies in the biomedical and life sciences 05/2012; 898:130-5. · 2.78 Impact Factor
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ABSTRACT: Shuang-huang-lian injection (SHLI) is a famous Chinese patent medicine, which has been wildly used in clinic for the treatment of acute respiratory tract infection, pneumonia, influenza, etc. The existing randomized controlled trial (RCT) studies suggested that SHLI could afford a certain anti-febrile action. However, seldom does research concern the pharmacological mechanisms of SHLI. In the current study, we explored plasma metabolomic profiling technique and selected potential metabolic markers to reveal the antipyretic mechanism of SHLI on yeast-induced pyrexia rat model using UPLC-Q-TOF/MS coupled with multivariate statistical analysis and pattern recognition techniques. We discovered a significant perturbance of metabolic profile in the plasma of fever rats and obvious reversion in SHLI-administered rats. Eight potential biomarkers, i.e. 1) 3-hydeoxybutyric acid, 2) leucine, 3) 16∶0 LPC, 4) allocholic acid, 5) vitamin B2, 6) Cys-Lys-His, 7) 18∶2 LPC, and 8) 3-hydroxychola-7, 22-dien-24-oic acid, were screened out by OPLS-DA approach. Five potential perturbed metabolic pathways, i.e. 1) valine, leucine, and isoleucine biosynthesis, 2) glycerophospholipid metabolism, 3) ketone bodies synthesis and degradation, 4) bile acid biosynthesis, and 5) riboflavin metabolism, were revealed to relate to the antipyretic mechanisms of SHLI. Overall, we investigated antipyretic mechanisms of SHLI at metabolomic level for the first time, and the obtained results highlights the necessity of adopting metabolomics as a reliable tool for understanding the holism and synergism of Chinese patent drug.PLoS ONE 01/2014; 9(6):e100017. · 3.73 Impact Factor