出 处:《中国病理生理杂志》2017年第4期661-668,共8页Chinese Journal of Pathophysiology
基 金:北京市自然科学基金资助项目(No.7102013);北京市中医药科技一般规划项目(No.JJ2013-02);北京市教委面上项目(No.KM20160025004)
摘 要:目的:探讨槲芪散是否通过调控Hedgehog信号通路抑制大鼠肝癌前病变的机制。方法:采用改良的Solt-Farber二步法复制大鼠肝癌前病变模型,于肝大部切除术后3 d开始给予槲芪散(8 g/kg)灌胃治疗,4周后收集血清和肝脏标本。分别采用HE染色、免疫组织化学、免疫荧光染色、Western blot、实时荧光定量PCR等方法,观察肝脏病理变化,检测肝组织谷胱甘肽S-转移酶π(glutathione S-transferase-π,GST-π)、甲胎蛋白(alpha-fetoprotein,AFP)、OV6、白蛋白(albumin,ALB)、Hedgehog信号分子Shh、Smo、Gli2及下游靶分子cyclin D、cyclin E的表达水平;利用比色法检测血清丙氨酸转氨酶(alanine aminotransferase,ALT)、天冬氨酸转氨酶(aspartate aminotransferase,AST)和γ-谷氨酰转移酶(gamma-glutamyltransferase,GGT)水平;给予Hedgehog信号通路抑制剂环杷明观察槲芪散在体外对肝卵圆细胞的影响。结果:与模型组比较,槲芪散可降低肝癌前病变大鼠血清ALT、AST和GGT水平,减轻肝癌前病变大鼠肝脏的病理变化,降低肝癌前病变标志物GST-π和AFP的表达;槲芪散可促使肝卵圆细胞标志物OV6和肝细胞标志物ALB的表达;槲芪散可激活肝脏Hedgehog信号通路,增加Shh、Smo、Gli2及其下游靶分子cyclin D、cyclin E的表达。结论:中药复方槲芪散可通过激活Hedgehog信号通路促进肝前体细胞增殖和诱导肝前体细胞向肝细胞分化,抑制肝癌前病变的形成,促进肝脏修复。AIM: To explore the influence of Huqi San on the Hedgehog signaling pathway in rats with prehepatocarcinoma. METHODS: The model of prehepatocarcinoma in the rats was established by a modified solt-farber method. The rats were intragastric administrated with Huqi San solution for 3 d after subtotal hepatectomy. Four weeks after administration of the Huqi San solution,the hepatic damage was observed by histopathological analysis. The protein expression of glutathione S-transferase-π( GST-π),alpha-fetoprotein( AFP),OV6,albumin( ALB) and glioma-associated oncogene homolog 2( Gli2) was detected by immunohistochemistry and immunofluorescence staining. The expression of Sonic hedgehog( Shh),Smoothened( Smo),Gli2,cyclin D and cyclin E at mRNA and protein levels in the rats was determined by RT-qPCR and Western blot,respectively. The serum levels of alanine aminotransferase( ALT),aspartate aminotransferase( AST) and gamma-glutamyltransferase( GGT) were assayed using diagnostic kits. RESULTS: Compared with model group,Huqi San decreased the serum levels of ALT,AST and GGT,and alleviated the pathological changes in prehepatocarcinoma rats. Huqi San inhibited the protein expression of GST-π and AFP( P〈0. 05) in the prehepatocarcinoma rats. Huqi San also promoted the protein expression of OV6 and ALB( P〈0. 05). Furthermore,Huqi San activated Hedgehog signaling pathway and its downstream targeting molecules such as Shh,Smo,Gli2,cyclin D and cyclin E. In addition,the results in vitro showed that Huqi San may activate Hedgehog signaling pathway and promoted oval cell proliferation. CONCLUSION: Huqi San not only promotes hepatic progenitor cell proliferation,but also induces hepatic progenitor cell differentiation and inhibits prehepatocarcinoma in the rats probably via activating Hedgehog signaling pathway.
关 键 词:肝癌前病变 HEDGEHOG信号通路 肝前体细胞 槲芪散
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