| Sanhuanglian mixture is developed to use in prevention and treatment of streptococcal disease in Tilapia.The present study was designed to study in vivo and in vitro anti-inflammatory effects of this formulation,and establish the quality standard and investigate the stability of this mixture.The anti-inflammatory effect of the mixture is detected by observing the inhibitory action on xylene-induced mouse ear edema model and the LPS-stimulated RAW264.7 cells.The results showed that the ear edema rate in mice treated with Sanhuanglian mixture at dose of 30 mL/kg body weight or 15 mL/kg body weight were 20.66%or 49.46%respectively and were significantly lower than negative control group(p<0.01).Besides thymus index in middle dose group,the index of liver,kidney,spleen and thymus in other experimental groups were not significantly different with the control group.The levels of NO,TNF-α,IL-1β,IL-6 and IL-8 in LPS-stimulated RAW264.7 cells were significantly decreased by Sanhuanglian mixture in a dose-dependent manner.These results showed that Sanhuanglian mixture had anti-inflammation effects both in vivo and in vitro without side effects and immune inhibition.Thin layer chromatography(TLC)used to identify the mixture.The results show Scutellaria baicalensis and fructus forsythiae image of TLC spots were clear,and specific spots were the same with the standard.This method can be used for quality control of the mixture.High performance liquid chromatography(HPLC)was used to analyse the main component of baicalin and forsythin in the mixture.The results showed that the method is specific,sensitive,reproducible,and can be used for the quality control of the mixture.According to the findings,baicalin content should be not less than 4.02mg/mL,forsythin content should be not less than 195.90 μg/mL.Sanhuanglian mixture is not sensitive to light by light acceleration test.Results of accelerated test show that the mixture is stable,in line with quality standard. |
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