苦参素穴位注射对肝纤维化防治及对肝组织金属蛋白酶组织抑制因子2表达影响

Prophylactic and Therapeutic Effects of Acupoint Injection with oxymatrine on Rat Liver Fibrosis and hepatic expression of tissue inhitor of metal protcase-2

目的:观察苦参素足三里(OM ZSL)注射预防及治疗大鼠肝纤维化的疗效及对肝组织中TIMP-2表达的影响。方法:采用四氯化碳诱导大鼠肝纤维化模型,SD大鼠35只,随机分为5组:正常对照组、模型对照组、苦参素腹腔注射(OM ip)对照组、OM ZSL注射治疗组、OM ZSL注射预防组。实验至10周末,测定血清ALT、AST、TBA、白蛋白(ALB)及肝组织中总蛋白(TP)水平。光镜、电镜观察肝细胞结构和肝纤维化程度;RT-PCR法检测肝组织中基质金属蛋白酶组织抑制因子-2(TIMP-2)mRNA的表达。结果:OM ZSL注射预防与治疗给药均能明显降低四氯化碳诱导肝纤维化大鼠ALT、AST、TBA含量,增加ALB及TP水平,(P<0.05,P<0.01)。病理学观察,OM ZSL注射防、治组大鼠肝组织病理改变明显减轻,假小叶结构明显减少;且OM ZSL预防组肝纤维化程度轻于治疗组。OM ZSL防、治组肝组织中TIMP-2 mRNA的表达显著低于模型对照组(P<0.05,P<0.01)。结论:OM ZSL注射对四氯化碳诱导的肝纤维化有较好的预防及治疗作用,其部分机制是通过保护肝细胞,减少TIMP-2 mRNA的表达,促进细胞外基质的降解,抑制细胞外基质沉积,从而减轻或逆转肝纤维化。

Objective：To investigate the prophylactic and therapeutic effects of oxymatrine on experimental liver fibrosis and hepatic expression of tissue inhitor of metal protcase-2. Methods ：The liver fibrosis model in rats was established by injecting carbon tetrachloride（ CCL4 ）. Thirty-five SD rats were randomly divided into normal control group, model control group, control group of oxymatrine intraperitoneal injection, treatment group of oxymatrine injection into zusanli（ZSL） and preventive group of oxymatrine injection into ZSL. At the end of the tenth week, The serum levels of alanine aminotransferase（ALT） ,aspartate aminotransferase（AST） , total bile acid （TBA） , albumin （ALB） , and total protein （TP） were measured. The histopathological changes were observed by light and electron microscopy. Reverse transcription polymerase chain reaction was used to detect the expressions of tissue inhibitor of metalloproteinase-2 （ TIMP-2 ） mRNA in liver tissue. Results ： In the treatment and preventive groups of oxymatrine injection into ZSL, the levels of serum ALT, AST,and TBA were markedly reduced, ALB,TP were increased markedly in liver fibrosis rats （P 〈 0.05,P 〈 0.01 ）. The pathological improvements were observed. In preventive and therapeutic groups of oxymatrine injection into ZSL, deposition of collageneous fiber in the liver tissue was markedly alleviated compared with model group and liver pseudoluboli reduced. The pathological studies showed that the liver fibrosis degree of oxymatrine ZSL preventive group was lighter than that of treatment group. The level of TIMP-2 mRNA in the liver were significantly reduced in preventive group and therapeutic group（ P 〈 0.01 ）. Conclusion：Oxymatrine injection in- to ZSL has obvious prophylactic and therapeutic effect on rats liver fibrosis induced by CCL4. Its antifibrotic mechanism was partly protecting hepatocyte,down- regulation on TIMP -2 mRNA expression , facilitating degradation of the extracellular matrix（ECM） and preventing ECM deposition in liver.