Department of Gastroenterology, The Second Hospital of Hebei Medical University, Hebei Key Laboratory of Gastroenterology, Hebei Institute of Gastroenterology, Hebei 050000, P.R. China.
Mol Med Rep. 2012 Dec;6(6):1231-8. doi: 10.3892/mmr.2012.1076. Epub 2012 Sep 11.
During the process of liver fibrosis, hepatic stellate cells (HSCs) play a critical role in the excessive production of extracellular matrix (ECM). Previous studies have indicated that the monomer IH764-3, one of the major bioactive components of Salvia miltiorrhiza, is able to inhibit HSC proliferation and induce the apoptosis of activated HSCs in vitro. In the current study, we used a rat model of liver fibrosis induced by bile duct ligation (BDL) to investigate the effect of the monomer IH764-3 on the induction of apoptosis in HSCs in vivo. The rat model of liver fibrosis was established by BDL. Immunohistochemical staining of α-smooth muscle actin (α-SMA) was performed to detect HSC activation and proliferation and HSC apoptosis was detected by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and α-SMA immunohistochemical double staining. In addition, the protein expression levels of focal adhesion kinase (FAK), p-FAK (Tyr397), extracellular signal-regulated kinase (ERK) and p-ERK and the mRNA expression levels of FAK and ERK were measured by western blotting and reverse transcription-polymerase chain reaction (RT-PCR), respectively. The monomer IH764-3 was associated with a significant decrease in intrahepatic fibrogenesis and collagen deposition and attenuated the liver fibrosis induced by BDL. Immunohistochemical staining revealed that the expression of α-SMA in the IH764-3 group was significantly decreased compared with that in the model group (12.92±2.45 vs. 22.65±2.16%, P<0.01). TUNEL and α-SMA immunohistochemical double staining also confirmed that IH764-3 increased the apoptotic rate of the activated HSCs (34.8±4.5 vs. 4.72±0.37%, P<0.01). Moreover, the results revealed that IH764-3 downregulated the expression levels of FAK, p-FAK (Tyr397), ERK and p-ERK in the liver tissue of rats with liver fibrosis. The monomer IH764-3 ameliorates experimental liver fibrosis by inhibiting HSC proliferation and inducing HSC apoptosis, warranting its use as a potential therapeutic agent in the treatment of liver fibrosis.
在肝纤维化过程中，肝星状细胞（HSCs）在细胞外基质（ECM）的过度产生中起着关键作用。先前的研究表明，丹参的主要生物活性成分之一 IH764-3 能够抑制 HSC 的增殖并在体外诱导活化的 HSC 凋亡。在本研究中，我们使用胆管结扎（BDL）诱导的大鼠肝纤维化模型来研究单体 IH764-3 在体内诱导 HSC 凋亡的作用。通过 BDL 建立大鼠肝纤维化模型。通过α-平滑肌肌动蛋白（α-SMA）免疫组织化学染色检测 HSC 的活化和增殖，通过末端脱氧核苷酸转移酶 dUTP 缺口末端标记（TUNEL）和α-SMA 免疫组织化学双重染色检测 HSC 凋亡。此外，通过 Western blot 和逆转录-聚合酶链反应（RT-PCR）分别测量了粘着斑激酶（FAK）、p-FAK（Tyr397）、细胞外信号调节激酶（ERK）和 p-ERK 的蛋白表达水平以及 FAK 和 ERK 的 mRNA 表达水平。单体 IH764-3 与肝内纤维发生和胶原沉积的显著减少有关，并减轻了 BDL 诱导的肝纤维化。免疫组织化学染色显示，IH764-3 组的α-SMA 表达明显低于模型组（12.92±2.45 对 22.65±2.16%，P<0.01）。TUNEL 和α-SMA 免疫组织化学双重染色也证实 IH764-3 增加了活化的 HSCs 的凋亡率（34.8±4.5 对 4.72±0.37%，P<0.01）。此外，结果表明 IH764-3 下调了肝纤维化大鼠肝组织中 FAK、p-FAK（Tyr397）、ERK 和 p-ERK 的表达水平。单体 IH764-3 通过抑制 HSC 增殖和诱导 HSC 凋亡来改善实验性肝纤维化，使其有望成为治疗肝纤维化的潜在治疗药物。
