Lee Shou-Lun, Hsu Wei-Hsiang, Tu Chia-Ming, Wang Wen-Han, Yang Cheng-Yao, Lee Hsien-Kuang, Chin Ting-Yu
Department of Biological Science and Technology, China Medical University, Taichung, Taiwan.
Department of Bioscience Technology, Chung Yuan Christian University, Taoyuan, Taiwan.
Evid Based Complement Alternat Med. 2021 Nov 12;2021:6065168. doi: 10.1155/2021/6065168. eCollection 2021.
The extract of freshwater clams has been used to protect the body against liver diseases in traditional folk medicine. This study aims at investigating the effects of freshwater clam extract on activated hepatic stellate cells (aHSCs), which are critical contributors to liver fibrosis.
The aHSCs used in this study were derived from hepatic stellate cells that were isolated and purified from the livers of male Wistar rats and then transformed into the activated phenotype by culturing on uncoated plastic dishes. Freshwater clam extract (CE) was collected after the outflow from the live freshwater clams in a water bath at 100°C for 60 min. The effects of CE on aHSCs were analyzed by MTT assay, flow cytometry, Oil Red O (ORO) staining, western blot, and real-time RT-PCR.
The results indicated that CE suppressed the proliferation of aHSCs through G0/G1 cell cycle arrest by downregulating cyclin D1 and upregulating p27. The expression levels of -SMA, collagen I, TGF-, and TNF- were inhibited in the CE-treated aHSCs. In addition, the CE treatment increased the lipid contents in aHSCs by promoting PPAR expression. Furthermore, CE modulated the expression of ECM-related genes, i.e., by upregulating MMP-9 and downregulating TIMP-II.
These data revealed that CE could induce the deactivation of aHSCs. We therefore suggest that CE has potential as an adjuvant therapeutic agent against hepatic fibrosis.
淡水蚌提取物在传统民间医学中已被用于保护机体免受肝脏疾病侵害。本研究旨在探讨淡水蚌提取物对活化肝星状细胞（aHSCs）的影响，活化肝星状细胞是肝纤维化的关键促成因素。
本研究中使用的aHSCs来源于从雄性Wistar大鼠肝脏中分离纯化的肝星状细胞，然后通过在未包被的塑料培养皿上培养转化为活化表型。在100°C水浴中对活淡水蚌进行60分钟流水处理后收集淡水蚌提取物（CE）。通过MTT法、流式细胞术、油红O（ORO）染色、蛋白质印迹法和实时逆转录聚合酶链反应分析CE对aHSCs的影响。
结果表明，CE通过下调细胞周期蛋白D1和上调p27使aHSCs的增殖停滞于G0/G1期，从而抑制其增殖。在CE处理的aHSCs中，α-SMA、I型胶原蛋白、转化生长因子-β和肿瘤坏死因子-α的表达水平受到抑制。此外，CE处理通过促进过氧化物酶体增殖物激活受体（PPAR）表达增加了aHSCs中的脂质含量。此外，CE调节细胞外基质（ECM）相关基因的表达，即通过上调基质金属蛋白酶-9（MMP-9）和下调金属蛋白酶组织抑制因子-II（TIMP-II）。
这些数据表明CE可诱导aHSCs失活。因此，我们认为CE有潜力作为抗肝纤维化的辅助治疗药物。
