苦蘵睡茄内酯提取物下调YAP和TGF-β/Smad通路抑制肝星状细胞激活改善小鼠肝纤维化  

The withanolide extract of Physalis angulata alleviates the liver fibrosis through regulation of YAP and TGF-β/Smad pathway

作　　者：杨挺 陈馨霖 祝小云 项德娟 罗建光[1] 张浩 YANG Ting;CHEN Xin-lin;ZHU Xiao-yun;XIANG De-juan;LUO Jian-guang;ZHANG Hao(Jiangsu Key Laboratory of Bioactive Natural Product Research,State Key Laboratory of Natural Medicines,School of Traditional Chinese Pharmacy,China Pharmaceutical University,Nanjing 211198,China)

机构地区：[1]中国药科大学中药学院,天然药物活性组分与药效国家重点实验室,江苏省天然活性物质发现与研究重点实验室,江苏南京211198

出　　处：《药学学报》2021年第11期2985-2994,共10页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(82074068,81872889)。

摘　　要：肝纤维化是慢性肝损伤的常见瘢痕反应,细胞外基质的过度积累是其主要特征,其中肝星状细胞的过度激活是肝纤维化发生的关键步骤。苦蘵睡茄内酯提取物(the withanolide extract of Physalis angulata, WEP)是从中药苦蘵(Physalis angulata L.)中提取富集的部位,主要含有酸浆苦素型的睡茄内酯类化合物。本实验通过建立四氯化碳及胆总管结扎诱导的小鼠肝纤维化模型,以及转化生长因子β1 (transforming growth factor-β1, TGF-β1)处理诱导的人肝星状细胞LX-2活化模型,结合不同浓度的WEP处理,研究WEP对肝纤维化的治疗作用,并探讨潜在的作用机制。本文中所有动物实验都获得中国药科大学伦理学委员会批准。动物实验结果显示,与模型组相比, WEP可降低血清中谷草转氨酶(aspartate transaminase, AST)和谷丙转氨酶(alanine transaminase, ALT)水平,减轻肝损伤,显著降低胶原沉积,改善肝纤维化水平。WEP对LX-2细胞无明显细胞毒性,但可显著降低肝纤维化标志物Ⅰ型胶原α1 (collagen typeⅠα1 chain, COL1A1)及α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)的mRNA和蛋白表达,抑制肝星状细胞活化。在体内外模型中, WEP可抑制yes相关蛋白(yes-associated protein, YAP)的表达及Smad家族蛋白2 (Smad family member 2, Smad2)的磷酸化水平。综上, WEP可通过下调YAP和TGF-β/Smad通路抑制肝星状细胞活化,在体内外表现出良好的抗肝纤维化活性。Liver fibrosis is a common scarring response to virtually all forms of chronic liver injury and is characterized by excessive accumulation of extracellular matrix. Excessive activation of hepatic stellate cells(HSCs) is a key step in liver fibrogenesis. The withanolide extract of Physalis angulata(WEP) is a physalin-type withanolide enriched partition from Physalis angulata. In this study, liver fibrosis mice models were established by CCl4 and bile duct ligation(BDL) in vivo. Transforming growth factor-β1(TGF-β1) was given in vitro to induce activation of human hepatic stellate cells LX-2 and treated with WEP at different concentrations. All animal experiments in this paper have been approved by the Ethics Committee of China Pharmaceutical University. The in vivo results showed that, compared with the CCl;or BDL group, WEP could reduce collagen deposition and liver damage, and reduce the levels of aspartate transaminase(AST) and alanine transaminase(ALT) in serum. In vitro,WEP had no significant cytotoxicity to LX-2 cells, but significantly reduced the mRNA and protein expressions of fibrotic markers collagen type Ⅰ α 1 chain(COL1 A1) and α-smooth muscle actin(α-SMA) and inhibited the activation of hepatic stellate cells. In addition, WEP inhibited the expression of yes-associated protein(YAP) and the phosphorylation level of Smad family member 2(Smad2) in vivo and in vitro. In conclusion, WEP can inhibit hepatic stellate cells activation via regulating the YAP and TGF-β-Smad pathways, and shows promising anti-fibrosis activity in vitro and in vivo.

关 键 词：肝纤维化 肝星状细胞激活 苦蘵睡茄内酯提取物 TGF-Β/SMAD YAP 

分 类 号：R966[医药卫生—药理学]