清肝方在ConA诱导的肝损伤中免疫调节作用的研究

Effects of QING GAN FANG on Immune-mediated Liver Injury Induced by ConA

目的:研究以清热利湿、疏肝解郁、柔肝缓急立法的清肝方对ConA诱导的免疫性肝损伤小鼠肝、脾、胸腺指数、转氨酶及病理改变的影响。方法:46只ICR雄性小鼠随机分成A(清肝方组)、B(清肝方加猪苓多糖腹腔注射组)、C(模型组)、D(正常对照组)四组。造模前,各组分别预治疗,其中,A组给予清肝方灌胃,B组予清肝方灌胃加腹腔注射猪苓多糖溶液,C、D组予蒸馏水灌胃,每日两次,共计5次,于末次给药后1h,采用0.2%的ConA溶液予小鼠尾静脉注射造模,分别于2h检测血清TNF-a,8h检测血清IFN-r、IL-4,10h用改良赖氏法检测血清中转氨酶的水平,并观察肝、脾病理变化情况。结果:A、B两组造模2h后,TNF-a水平明显降低;8h后血清IFN-r升高、IL-4水平降低;10h后,可明显降低肝、脾指数及血清转氨酶水平。组织结构分析表明清肝方及清肝方加猪苓多糖腹腔注射组,可明显减轻肝脏及脾脏的病理损伤。结论:清肝方通过激活细胞免疫,增强清除病毒的能力,发挥扶正祛邪之功效,同时,通过减少TNF-a,抑制NF-kB信号转导途径,减少后续炎症因子的释放,减轻ConA诱导的肝的免疫性损伤。

Objective：To investigate the effect of QING GAN FANG on the serum ALT. AST and pathologica -1 changes in ConA- induced hepatitis.Methods：46 1CR mice were randomly divided into four groups （A- QING GAN FANG group, B- QING GAN FANG together with polyporus polysaccharide （PPS） administered intraperitone -ally rote group, C-model group and D-normal group）, Among the four groups, mice were fed with QING GAN FANG. QING GAN FANG together with PPS administered intraperitoneally into, distilled water, distilled water in order,twice a day. add up to 5 times. 1 hour after medical treatments,ConA was injected intravenously at a dose of 20 mg/kg body weight to induce hepatitis. Serum concentrations of TNF-a were determined using specific enzyme -linked immunosorbent assay kits after 2 hours, both IFN-r and IL-4 were determin -ed after 8 hours in the same way; After 10 hours, the activities of liver enzymes aspartate transaminase （AST） and alanine transaminase （ALT） in the serum were determined, and liver slice were Stained with hematoxylin and eosin （H＆E）.Results： Compared to ConA group . both A and B groups resulted in significant suppression of TNF-α, IL-4,but activation of IFN- r. moreover, it could significantly decrease AST, ALT levels and prevent against liver injury at 10 hours after ConA challeng -e .Conclusions： QING GAN FANG could effectively protect hepatocyte from immune injury in ConA-induced hepatitis, by way of activating cell-mediated immtmity to clear the virus, and reducing the release of TNF-α to suppress the activation of NF-KB, thereby reducing the subsequent release of inflammatory factors.