扶正化瘀胶囊抑制BALB/C小鼠血吸虫病肝纤维化的机制

Experimental study on the mechanism of action of Fuzheng huayu recipe on Schistosoma japanicum liver fibrosis in BALB/C Mice

目的:观察扶正化瘀胶囊治疗日本血吸虫病鼠早期肝纤维化的疗效并初步探讨相关机制。方法:将45只BALB/C小鼠随机分为对照组、感染组、治疗组3组,每组各15只小鼠,采用日本血吸虫尾蚴(20±1)条/只,经腹部皮肤感染BALB/C小鼠建立血吸虫病肝纤维化模型,治疗组给予扶正化瘀方4.6 g.(kg.d)-1,灌胃,qd,疗程8周。HE和Masson染色观察肝组织病理学改变,免疫组化检测转化生长因子β1(TGF-β1)、α平滑肌动蛋白(α-SMA)和血管紧张素Ⅱ1型受体(AT1R)在肝组织的表达,实时荧光定量(RT-PCR)检测检测TGF-β1、α-SMA和AT1R的mRNA水平。结果:与感染组相比,治疗组小鼠肝脏组织病理损害减轻,肝脏的胶原面积、虫卵肉芽肿结节面积分别为(0.15±0.02)mm2和(0.12±0.04)mm2,比感染组小鼠肝脏的胶原面积(0.29±0.03)mm2和虫卵肉芽肿结节面积(0.23±0.06)mm2均明显减小(P<0.05),免疫组化及RT-PCR均显示治疗组小鼠肝脏α-SMA、TGF-β1、AT1R表达显著低于感染组(P均<0.05)。结论:扶正化瘀胶囊可抑制小鼠鼠血吸虫病肝纤维化程度,其作用机制可能与其下调AT1R表达,抑制TGF-β1、α-SMA的转录,减少胶原沉积有关。

OBJECTIVE To explore the effects of Fuzheng huayu recipe on schistosoma japanicum liver fibrosis and investi gate its of mechanism of action involving in Schistosorna japanicom liver fibrosis. METHODS 45 BALB/C mice were divided into 3 groups, normal group （n = 15）, infected group （n = 15） and treated group （n = 15）. The infected group and treated group were infected by （20 ± 1） cercarie of Schistosoma japonicum per mouse. 8 weeks later, the schistosomal liver fibrosis models were successfully induced. The mices of treated group were treated by intragastric administration with Fuzheng huayu recipe 4. 6 g,（kg.d） for 8 weeks. After 8 weeks of treatment, hepatic pathological changes and collagen fibers deposition were observed by HE staining and Masson staining. Immunohistochemistry was used to measure the expression and location of msmooth muscle actin （mSMA）, transforming growth factor-beta 1 （TGF-β1） and angiotensin ± type 1 receptor（AT1R）. The expression levels of mSMA mRNA, TGF-β1 mRNA and AT1R mRNA were measured by quantitative RT PCR. RESULTS Compared with the infected group, liver pathological damage in treated group were reduced. Area of collagen liver and area of eggs granulomatous nodule in the treated group （0. 15± 0. 02） mm2 and （0. 12 ± 0. 04） mm2 , respectively） was significantly less than those in the infected group （0. 29 ± 0. 03） mm2 （P〈0. 05） ; （0.23 ± 0. 06） mm2 （P〈0. 05）, respectively）. Immuno histochemistry and RT-PCR showed that in the treated group α-SMA, TGF β1, AT1R expression in the liver tissue was signif icantly lower than that of the infected group （P〈0. 05）. CONCLUSION Fuzheng huayu recipe may inhibit liver fibrosis pro gression due to schistosoma japanicum in BALB/C mice and its mechanism may be associated with downregulation ATIR ex- pression, inhibiting of transcription of TGF-β1, α-SMA, and then reduction of collagen deposition.