壳聚糖中药复方防治非酒精性脂肪性肝炎的实验研究

目的探明壳聚糖中药复方拮抗非酒精性脂肪性肝炎(NASH)的作用及其机制。方法健康雄性C57BL/6J小鼠18只,随机分为3组,每组6只。采用高脂、胆碱-蛋氨酸缺乏(high fat,methionine and choline deficient diet,MCD)饮食4周建立小鼠NASH模型,应用壳聚糖中药复方进行干预实验,并以胆碱-蛋氨酸充足饮食设立对照组。酶法测定血清丙氨酸氨基转移酶(alanine aminotransferase,ALT)、总胆固醇(total cholesterol,TC)和三酰甘油(triglyeride,TG)水平;双抗体夹心ELISA法测定血清肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和白介素-6(interleukin-6,IL-6)水平;光镜下观察肝组织脂肪变程度、炎症活动及纤维化程度;实时定量PCR法检测肝组织TNF-α和IL-6mRNA的表达。结果对照组小鼠肝脏未出现明显的脂肪变及炎症;模型组小鼠肝脂肪变及炎症明显,TNF-α和IL-6的表达显著升高(P<0.01);壳聚糖中药复方干预组肝组织脂肪变及炎性均较模型组明显减轻,TNF-α和IL-6的表达明显降低(P<0.01)。结论壳聚糖中药复方可通过抑制炎症因子TNF-α和IL-6的表达、纠正脂代谢紊乱来阻止或延缓非酒精性脂肪性肝炎的进展,是防治脂肪性肝病的有效措施之一。

Experimental study on the mechanism of chitosan recipe in preventing and treating non-alcoholic steatohepatitis

Objective To investigate the potential effect and molecular mechanism of chitosan recipe in preventing and treating non-alcoholic steatohepatitis(NASH).Methods Eighteen healthy male C57BL/6J mice were randomly divided into three groups:control group,model group and intervention group,with 6 mice in each group.The mice in control group were fed with methionine-choline-sufficient diet,and the mice in model group and intervention group were fed with high fat and methionine-choline-deficient(MCD)diet for 4 weeks to induce NASH,besides,the mice in intervention group were administrated with chitosan recipe solution(0.7g/kg)by daily intragastric gavag.Four weeks later,all the mice were sacrificed,and the serum levels of alanine aminotransferase(ALT),total cholesterol(TC)and triglyceride(TG)were detected by using automatic biochemistry analyzer.The serum levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)were measured by double antibody sandwich Enzyme-linked immunosorbent assay(ELISA).The histological changes were observed under light microscope.The mRNA expressions of TNF-αand IL-6 were detected by real-time quantitative polymerase chain reaction(RQ-PCR).Results As compared with the mice in control group,the mice in model group developed obvious steatohepatitis with hepatocyte ballooning changes,and the levels of TNF-αand IL-6 were significantly increased(P<0.01),furthermore,these pathological changes in intervention group were significantly lighter than those in model group,and the levels of TNF-αand IL-6 were significantly decreased(P<0.01).Conclusion Chitosan recipe can prevent and delay the progression of NASH by inhibiting the expressions of TNF-αand IL-6 and correcting lipid disorders,which is one of the effective measures for preventing and treating fatty hepatic diseases.