三七总皂苷对肝纤维化大鼠基质金属蛋白酶-13及其抑制因子-1表达的影响

Effect of Panax notoginseng saponins on expressions of MMP-13 and TIMP-1 in rats with hepatic fibrosis

目的:观察三七总皂苷(PNS)对四氯化碳(CCl4)诱导的肝纤维化大鼠基质金属蛋白酶(MMP)-13、基质金属蛋白酶抑制因子(TIMP)-1表达的影响,探讨PNS抗肝纤维化作用的可能机制。方法:将大鼠随机分成正常组、模型组、PNS低、中、高剂量(50,100,200 mg.kg-1)组和秋水仙碱(Col,0.1 mg.kg-1)组。除正常组外其余各组采用CCl4皮下注射的方法诱导肝纤维化大鼠模型,每周2次,连续18周。于造模第9周起,给药组分别灌胃相应的受试药物,正常组和模型组灌胃等容量的生理盐水,疗程10周。实验结束后,计算肝脏、脾脏指数;比色法测定血清中ALT,AST含量;同时取固定部位肝脏组织,HE染色观察肝组织病理学改变,免疫组化技术测定肝组织中MMP-13,TIMP-1蛋白的表达,RT-PCR技术测定MMP-13,TIMP-1mRNA的表达。结果:与模型组相比,三七总皂苷(100,200 mg.kg-1)不仅可减轻大鼠肝纤维化程度,降低肝脏、脾脏指数及血清ALT,AST的含量,还可显著升高肝纤维化大鼠肝脏MMP-13的表达,降低TIMP-1的表达。结论:三七总皂苷对大鼠肝纤维化具有一定的保护作用,其机制可能与上调MMP-13,抑制TIMP-1的表达,促进胶原降解有关。

Objective： To investigate the effect of Panax notoginseng saponins （PNS） on the expressions of matrix metallopro- teinase （MMP） -13 and its tissue inhibitor of metalloproteinase （TIMP） -1 in carbon tetrachloride （ CC14 ） -induced hepatic fibrosis rats, and explore the possible mechanism of PNS＇s effect against hepatic fibrosis. Method： The rats were randomly divided into 6 groups： the normal group, the model group, PNS （50, 100, 200 mg ·kg^-1 ） treatment groups and the Col （0. 1 mg·kg^-1 ） group. Apart from the normal group, all of the remaining groups were subcutaneously injected with CC14 twice a week for 18 weeks, in order to estab- lish the hepatic fibrosis rat model. Since the 9th weeks, each treatment group was orally administered with corresponding drugs, and the normal group and the model group were orally administered with equal volume of normal saline for 10 weeks. After the end of the exper- iment, liver and spleen indexes were calculated ; the levels of serum ALT and AST were measured by chromatometry. Liver tissues were collected to detect the pathological alteration HE staining; protein expressions of MMP-13 and TIMP-1 were determined with immunino- chemistry. Moreover, MMP-13 and TIMP-1 mRNA expressions was detected by RT-PCR technology. Result： Compared with the model group, PNS （100, 200 mg·kg^-1） significantly mitigated hepatic fibrosis in rats, reduced liver and spleen indexes, ALT and AST contents in serum, and TIMP-1 expression, and notably increased MMP-13 expression in rats with hepatic fibrosis. Conclusion： P. notoginseng saponins have certain protective effect in rats with hepatic fibrosis. Its mechanism is related to up-regulating MMP-3, inhibiting TIMP-1 expression and improving collagen degradation.