Feng Li-Li, Cao Wen-Fu
Department of Integration of Traditional Chinese Medicine and Western Medicine, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
Zhongguo Zhong Yao Za Zhi. 2017 Mar;42(5):964-969. doi: 10.19540/j.cnki.cjcmm.20170121.015.
To explore the mechanism of Ezhu-containing serum in inhibiting the expression of sonic hedgehog(Shh) and glioma-associated oncogene homolog-1(Gli1) in hepatic stellate cells(HSCs) induced by leptin. Twenty sprague-dawley (SD) rats were randomly divided into 2 groups (n=10), and given Ezhu-decoction and physiological saline by gavage for 10 days to prepare drug-containing serums. The HSCs during the exponential growth phase were divided into 7 groups： blank control group, model group, hedgehog pathway inhibitor(cyclopamine) group, Ezhu group, Ezhu and cyclopamine group, hedgehog pathway agonost(pumorphamine) group, Ezhu and purmorphamine group. HSCs were cultured in vitro and induced with 100 μg•L ⁻¹ leptin(except for the blank control group), then treated separately with the corresponding drugs for 24 hours. After the cells were collected, HSCs proliferation was detected using MTT colorimetric assay; the expressions of Shh and Gli1 were determined by PT-PCR, Western blot and immunofluorescence, respectively. The expressions of Shh and Gli1 were significantly increased after the HSCs of rats were induced by leptin (compared with the blank control group, P<0.01). After being interfered with Hh pathway inhibitor (cyclopamine) and Ezhu-containing serum, the expressions of Shh and Gli1 were decreased significantly(compared with the model group, P<0.01). After Ezhu-containing serum was used to interfere the Hh pathway inhibitor group, the mRNA and protein expressions of Shh and Gli1 were decreased significantly(compared with the model group, P<0.01). After Ezhu-containing serum was used to interfere the purmorphamine group, the mRNA and protein expressions of Shh and Gli1 decreased significantly(compared with the purmorphamine group, P<0.01). Ezhu-containing serum plays an important role in inhibiting HSCs activation by taking part in hedgehog signaling pathway, so as to regulate the expression of Shh and Gli1 in leptin-induced HSCs and then inhibit liver fibrosis.
探讨含莪术血清抑制瘦素诱导的肝星状细胞（HSCs）中声波刺猬因子（Shh）和胶质瘤相关癌基因同源物1（Gli1）表达的机制。将20只Sprague-Dawley（SD）大鼠随机分为2组（n = 10），分别给予莪术水煎剂和生理盐水灌胃10天制备含药血清。将指数生长期的HSCs分为7组：空白对照组、模型组、刺猬信号通路抑制剂（环杷明）组、莪术组、莪术与环杷明组、刺猬信号通路激动剂（普莫法明）组、莪术与普莫法明组。体外培养HSCs并用100μg•L⁻¹瘦素诱导（空白对照组除外），然后分别用相应药物处理24小时。收集细胞后，采用MTT比色法检测HSCs增殖；分别通过PT-PCR、Western blot和免疫荧光法检测Shh和Gli1的表达。大鼠HSCs经瘦素诱导后，Shh和Gli1的表达显著增加（与空白对照组相比，P<0.01）。经Hh信号通路抑制剂（环杷明）和含莪术血清干预后，Shh和Gli1的表达显著降低（与模型组相比，P<0.01）。含莪术血清干预Hh信号通路抑制剂组后，Shh和Gli1的mRNA和蛋白表达显著降低（与模型组相比，P<0.01）。含莪术血清干预普莫法明组后，Shh和Gli1的mRNA和蛋白表达显著降低（与普莫法明组相比，P<0.01）。含莪术血清通过参与刺猬信号通路在抑制HSCs激活中起重要作用从而调控瘦素诱导的HSCs中Shh和Gli1的表达进而抑制肝纤维化。
