蒙药额力根-7对肝纤维化的改善作用及机制研究
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篇名: 蒙药额力根-7对肝纤维化的改善作用及机制研究
TITLE: Study on Improvement Effects of Mongolian Medicine Eligen- 7 on Hepatic Fibrosis and Its Mechanism
摘要: 目的:研究蒙药额力根-7对肝纤维化的改善作用及机制。方法:以大鼠肝星状细胞HSC-T6为研究对象,将细胞分为模型组(空白血清)和额力根-7含药血清低、中、高剂量组(10%、15%、20%额力根-7含药血清)。各组细胞均先加入转化生长因子β溶液(0.2mg/mL)培养48h以复制肝纤维化模型,然后再加入相应空白或含药血清。检测各组细胞的光密度(OD)值并计算增殖抑制率(加血清后培养24、48、72h时);检测各组细胞的凋亡率、周期分布和细胞中Ⅰ型胶原蛋白(CollagenⅠ)、α-平滑肌肌动蛋白(α-SMA)以及磷脂酰肌醇-3激酶/蛋白激酶B(PI3K/Akt)信号通路相关因子[PI3K、Akt、磷酸酯酶与张力蛋白同源物(PTEN)]的mRNA和蛋白表达水平(加血清后培养24h时)。进一步以Wistar大鼠为研究对象,将大鼠分为空白组、模型组和额力根-7低、中、高剂量组(135、270、405mg/kg),每组10只。空白组和模型组大鼠灌胃0.5%羧甲基纤维素钠溶液,其余各组大鼠均灌胃相应药物,每天1次,连续10周。末次灌胃后,观察大鼠肝组织病理形态学变化,检测肝组织中CollagenⅠ、α-SMA、PI3K、Akt、PTEN的mRNA和蛋白表达水平。结果:与模型组比较,各剂量给药组细胞的OD值(培养72h时的中、高剂量组除外)、S期细胞比例均显著降低(P<0.01),晚期凋亡率、早期凋亡率(低剂量除外)、总凋亡率和G2/M期细胞比例均显著升高(P<0.01);细胞和大鼠肝组织中CollagenⅠ、α-SMA、PI3K、Akt的mRNA和蛋白表达水平均显著降低(P<0.05或P<0.01),PTEN的mRNA和蛋白表达水平均显著升高(P<0.05或P<0.01)。结论:额力根-7具有抗肝纤维化作用,其作用机制可能与调控PI3K/Akt信号通路活性、促进肝星状细胞凋亡有关。
ABSTRACT: OBJECTIVE:To study the improv ement effects of Mongolian medici ne eligen- 7 on hepatic fibrosis (HF)and its mechanism. METHODS :Taking rat hepatic stellate cells HSC-T 6 as research object ,the cells were divided into model group (blank serum )and low-dose ,medium-dose and high-dose groups of eligen- 7 containing serum (10%,15% and 20% eligen-7 containing serum ). Transforming growth factor β solution(0.2 mg/mL)was added into the cells for 48 h to induce liver fibrosis model,and then added into the corresponding blank or drug-contained serum. The optical density (OD)of cells in each group was measured and inhibition rate of cell proliferation was calculated (after treated for 24,48 and 72 h). The apoptotic rate and cycle distribution of cells were detected ;mRNA and protein expression of type Ⅰ collagen(Collagen Ⅰ),α-smooth muscle actin (α-SMA),phosphatidylinositol-3-kinase and protein-serine-threonine kinase (PI3K/Akt)signaling pathway related factors (PI3K, Akt,PTEN)were also detected (after treated for 24 h). Wistar rats were further divided into blank group ,model group and low-dose,medium-dose and high-dose groups of eligen- 7(135,270,405 mg/kg),with 10 rats in each group. Blank group and model group were given 0.5% sodium carboxymethyl cellulose solution intragastrically ;other groups were given relevant medicine intragastrically,once a day ,for consecutive 10 weeks. After last intragastric administration ,the pathomorphological changes of liver tissue were observed ;mRNA and protein expression of Collagen Ⅰ,α-SMA,PI3K,Akt and PTEN were detected in liver tissue. RESULTS :Compared with model group ,OD value (except for medium-dose and high-dose groups of eligen- 7 containing serum)and the proportion of cells at S phase in administration groups were decreased significantly (P<0.01);late apoptotic rate , early apoptotic rate (except for low-dose group ),total apoptotic rate and the proportion of cells at G 2/M phase increased significantly(P<0.01);mRNA and protein expression of Collagen Ⅰ,α-SMA,PI3K and Akt in cells and liver tissue were decreased significantly (P<0.05 or P<0.01),those of PTEN were increased significantly (P<0.05 or P<0.01). CONCLUSIONS : Eligen-7 shows the effect of anti-hepatic fibrosis , themechanism of which may be related to regulating the activity of PI 3K/Akt signaling pathway and promoting the apoptosis of hepatic stellate cells.
期刊: 2021年第32卷第23期
作者: 颜羽昕,张春艳,高晓阳,马月宏
AUTHORS: YAN Yuxin,ZHANG Chunyan ,GAO Xiaoyang ,MA Yuehong
关键字: 肝纤维化;额力根-7;PI3K/Akt信号通路;细胞凋亡;肝星状细胞;大鼠
KEYWORDS: Hepatic fibrosis ;Eligen-7;PI3K/Akt signaling
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