原阿片碱对人肝星状细胞HSC-LX2增殖的抑制作用及机制研究
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篇名: 原阿片碱对人肝星状细胞HSC-LX2增殖的抑制作用及机制研究
TITLE:
摘要: 目的:考察原阿片碱对人肝星状细胞HSC-LX2增殖的抑制作用,并对其机制进行初步探讨。方法:采用MTT法测定25、50、100、200、400、500 μmol/L原阿片碱作用24 h对HSC-LX2细胞增殖的影响,计算细胞增殖抑制率。另取HSC-LX2细胞分为对照组(含5%胎牛血清的1640培养基)和原阿片碱低、中、高浓度组(100、200、400 μmol/L),作用24 h后,流式细胞术测定细胞凋亡率;荧光定量逆转录-聚合酶链式反应(RT-PCR)法测定细胞中α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原蛋白(Collagen Ⅰ)、Ⅲ型胶原蛋白(Collagen Ⅲ)、基质金属蛋白酶2(MMP-2)、组织金属蛋白酶抑制剂1(TIMP-1) mRNA的相对表达量,Western blot法测定细胞中α-SMA、Collagen Ⅰ、Collagen Ⅲ、MMP-2蛋白的相对表达量。结果:25、50、100、200、400、500 μmol/L原阿片碱对HSC- LX2细胞的增殖抑制率分别为0、6.9%、18.7%、34.2%、48.9%、53.9%。与对照组比较,原阿片碱低、中、高浓度组细胞中Collagen Ⅰ、TIMP-1 mRNA相对表达量和α-SMA蛋白相对表达量均显著降低,MMP-2蛋白相对表达量显著升高,差异均有统计学意义(P<0.05或P<0.01);原阿片碱中、高浓度组细胞的凋亡率和MMP-2 mRNA相对表达量显著升高,α-SMA、Collagen Ⅲ mRNA相对表达量和Collagen Ⅰ、Collagen Ⅲ 蛋白相对表达量均显著降低,差异均有统计学意义(P<0.05或P<0.01)。 结论:原阿片碱可抑制HSC-LX2细胞增殖,诱导其凋亡,可降低α-SMA、Collagen Ⅰ、Collagen Ⅲ、TIMP-1表达,升高MMP-2表达有关。
ABSTRACT: OBJECTIVE: To investigate inhibitory effects of protopine on the proliferation of human hepatic stellate cells HSC-LX2 and to explore its mechanism preliminarily. METHODS: MTT assay was used to detect the effects of 25, 50, 100, 200, 400 and 500 μmol/L protopine on the proliferation of HSC-LX2 cells. The inhibitory effect of cell proliferation was calculated. HSC-LX2 cells were divided into control group (1640 medium containing 5% fetal bovine serum), protopine low-concentration, medium-concentration and high-concentration groups (100, 200, 400 μmol/L). After treated for 24 h. The apoptotic rate of the cells was detected by flow cytometry. RT-PCR was used to determine the mRNA expression of α-SMA, Collagen Ⅰ, Collagen Ⅲ, MMP-2 and TIMP-1 in cells. The protein expressions of α-SMA, Collagen Ⅰ, Collagen Ⅲ and MMP-2 were detected by Western blot. RESULTS: The inhibitory rates of 25, 50, 100, 200, 400 and 500 μmol/L protopine on proliferation HSC-LX2 cells were 0, 6.9%, 18.7%, 34.2%, 48.9%, 53.9%, respectively. Compared with control group, mRNA expression of Collagen Ⅰ, TIMP-1 and protein expression of α-SMA were decreased significantly in protopine low-concentration, medium-concentration and high-concentration groups, while protein expression of MMP- 2 was increased significantly, with statistical significance (P<0.05 or P<0.01). Apoptotic rate of HSC-LX2 cells and mRNA expression of MMP-2 were increased significantly in protopine medium-concentration and high-concentration groups, mRNA expression of α-SMA and Collagen Ⅲ, protein expression of Collagen Ⅰ and Collagen Ⅲ were decreased significantly, with statistical significance (P<0.05 or P<0.01). CONCLUSIONS: Protopine can induce the apoptosis of HSC-LX2 cells and inhibit their cell proliferation, and reduce the expression of a-SMA, Collagen Ⅰ, Collagen Ⅲ and TIMP-1, and increase the expression of MMP-2.
期刊: 2019年第30卷第23期
作者: 吴新玉,李靖,张金娟,廖尚高,梅青,吴亚云,席晓岚
AUTHORS: WU Xinyu,LI Jing,ZHANG Jinjuan,LIAO Shanggao,MEI Qing,WU Yayun,XI Xiaolan
关键字: 原阿片碱;肝星状细胞HSC-LX2;增殖;凋亡;机制
KEYWORDS: Protopine; Hepatic stellate cells HSC-LX2; Proliferation; Apoptosis; Mechanism
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