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目的观察山奈酚(Kaempferol,KA)对肝组织转化生长因子(TGF)-β1/Smads信号通路的影响,探讨其预防血吸虫病肝纤维化的作用机制。方法 40只BALB/c小鼠随机分为正常对照组(8只)、吡喹酮组(8只)、吡喹酮+山奈酚4个剂量组(5、10、15、20 mg/kg)(每组6只)。除正常对照组外,其余5组小鼠同时感染日本血吸虫,感染6周后吡喹酮组及吡喹酮+山奈酚4个剂量组给予吡喹酮连续灌胃2 d,其后吡喹酮+山奈酚4个剂量组分别以5、10、15、20 mg/kg山奈酚连续灌胃6周,吡喹酮组予以等量生理盐水灌胃6周。干预结束后处死小鼠并取肝脏,以苏木精-伊红(HE)和Masson胶原纤维染色法观察小鼠肝虫卵肉芽肿面积和肝纤维化程度;以免疫组化染色观察小鼠肝组织TGF-β1、Smad2/3、Smad7蛋白的表达情况;采用实时荧光定量PCR检测肝组织Smad2/3、Smad7 mRNA的表达情况。结果与吡喹酮组小鼠相比,吡喹酮+山奈酚4个剂量组肝脏虫卵肉芽肿面积较小,纤维化程度较轻,肝组织Smad2/3蛋白及其mRNA表达水平较低,Smad7蛋白及其mRNA表达增加,差异均有统计学意义(P均<0.05)。结论山奈酚通过减少肝组织内TGF-β1、Smad2/3的表达及增加Smad7的表达,能明显减轻吡喹酮杀虫治疗后虫卵肉芽肿所致肝纤维化。
Objective To investigate the effect of Kaempferol (KA) on the expression of transforming growth factor (TGF) -β1 / Smads in liver tissue and its mechanism of action in preventing liver fibrosis induced by schistosomiasis. Methods 40 BALB / c mice were randomly divided into normal control group (n = 8), praziquantel group (n = 8), praziquantel + kaempferol four dose groups (5,10,15,20 mg / kg) (6 per group). In addition to the normal control group, the other 5 groups of mice were infected with Schistosoma japonicum at the same time. After 6 weeks of infection, praziquantel and praziquantel plus kaempferol were administered continuously for 2 days. Praziquantel + Kaempferol 4 doses were 5, 10, 15, 20 mg / kg kaempferol continuous gavage for 6 weeks, praziquantel group was given normal saline for 6 weeks. After the intervention, the mice were sacrificed and the livers were sacrificed to observe the granuloma area and the degree of liver fibrosis in mice by hematoxylin-eosin (HE) and Masson collagen staining. The expression of hepatic fibrosis was observed by immunohistochemistry The expression of TGF-β1, Smad2 / 3 and Smad7 proteins were detected by real-time fluorescence quantitative PCR. The expressions of Smad2 / 3 and Smad7 mRNA in liver tissues were detected by real-time fluorescence quantitative PCR. Results Compared with the praziquantel group, the four groups of praziquantel and kaempferol had smaller egg granuloma and lighter fibrosis, lower expression of Smad2 / 3 protein and mRNA, Smad7 protein and mRNA expression increased, the differences were statistically significant (P all <0.05). Conclusion Kaempferol could significantly reduce hepatic fibrosis induced by egg granuloma after praziquantel treatment by decreasing the expression of TGF-β1 and Smad2 / 3 and increasing the expression of Smad7 in liver tissue.