白屈菜碱对活化后大鼠肝星状细胞CFSC-8B的增殖、胶原合成及TGF-β1受体的影响
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篇名: 白屈菜碱对活化后大鼠肝星状细胞CFSC-8B的增殖、胶原合成及TGF-β1受体的影响
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摘要: 目的:考察白屈菜碱对活化后大鼠肝星状细胞CFSC-8B的增殖、胶原合成及转化生长因子β1(TGF-β1)受体的影响。方法:取对数生长期的CFSC-8B细胞,分为正常对照组、模型组、溶剂组(乙醇)、阳性对照组(1 μg/mL秋水仙碱乙醇溶液)和白屈菜碱低、中、高浓度组(2.1、4.2、8.4 μg/mL白屈菜碱乙醇溶液)。除正常对照组外,其余各组细胞利用20 μg/L的TGF-β1作用24 h进行活化,后5组细胞给予相应药物干预24 h。采用CCK-8法检测各组细胞增殖情况,酶消化法检测细胞上清液中羟脯氨酸(Hyp)含量,酶联免疫吸附测定法(ELISA)检测细胞上清液中Ⅰ型胶原蛋白(Col-Ⅰ)和Ⅲ型胶原蛋白(Col-Ⅲ)水平,Western blot法检测细胞中TGF-β1Ⅰ型受体(TβR-Ⅰ)和TβR-Ⅱ蛋白的表达,实时荧光定量-聚合酶链式反应法检测细胞中α-平滑肌肌动蛋白(α-SMA)、TβR-Ⅰ和TβR-ⅡmRNA的表达。结果:与正常对照组比较,模型组细胞增殖率、Hyp含量、Col-Ⅰ和Col-Ⅲ水平、TβR-Ⅰ和TβR-Ⅱ的蛋白表达水平以及α-SMA、TβR-Ⅰ、TβR-Ⅱ的mRNA表达水平均显著升高(P<0.05)。与模型组比较,溶剂组细胞的上述指标均无明显差异(P>0.05);白屈菜碱低浓度组细胞增殖率无明显差异(P>0.05);阳性对照组和白屈菜碱高浓度组细胞的上述指标均显著降低(P<0.05);白屈菜碱中浓度组细胞除Hyp和Col-Ⅲ水平降低不显著外,上述其余指标均显著降低(P<0.05)。与白屈菜碱中浓度组比较,白屈菜碱高浓度组细胞增殖率、Col-Ⅰ水平、TβR-Ⅰ和TβR-Ⅱ的蛋白及mRNA表达水平均显著降低(P<0.05)。结论:白屈菜碱能抑制活化后CFSC-8B细胞的增殖、胶原合成以及TβR-Ⅰ、TβR-Ⅱ蛋白和mRNA的表达。
ABSTRACT: OBJECTIVE: To investigate the effects of chelidonine on proliferation, collagen synthesis and TGF-β1 receptor of activated hepatic stellate cells CFSC-8B. METHODS: CFSC-8B cells in logarithmic phase were collected and then divided into normal control group, model group, solvent group (ethanol), positive control group (1 μg/mL colchicine ethanol solution), chelidonine low, medium and high concentration groups (2.1, 4.2, 8.4 μg/mL chelidonine ethanol solution). Except for normal control group, other groups were activated with 20 μg/L TGF-β1 for 24 h; the latter 5 groups were intervened with relevant medicine for 24 h. Cell proliferation of activated cells was assayed by CCK-8 assay. Hydroxyprolin (Hyp) content was assayed by enzyme digestion; the levels of typeⅠ collagen (Col-Ⅰ) and type Ⅲ collagen (Col-Ⅲ) were assayed by ELISA; the expressions of TβR-Ⅰ and TβR-Ⅱ protein were assessed by Western blot; mRNA expressions of α-SMA, TβR-Ⅰ and TβR-Ⅱ in hepatic stellate cells were assessed by RT-PCR. RESULTS: Compared with normal control group, cell proliferation rate, Hyp content, the levels of Col-Ⅰ and Col-Ⅲ, the protein expressions of TβR-Ⅰ and TβR-Ⅱ as well as mRNA expressions of α-SMA, TβR-Ⅰ and TβR-Ⅱ were increased significantly (P<0.05). Compared with model group, there were no significant difference in above indexes of hepatic stellate cells in solvent group (P>0.05); there were no significant difference in the proliferation rate of hepatic stellate cells in chelidonine low concentration group (P>0.05), above indexes of hepatic stellate cells were decreased significantly in positive control group and chelidonine high concentration group (P<0.05). The decrease of Hyp and Col-Ⅲ levels were not significant in chelidonine medium concentration, but other above indexes were decreased significantly (P<0.05). Compared with chelidonine medium concentration group, the rate of cell proliferation, Col-Ⅰ level, protein and mRNA expressions of TβR-Ⅰ and TβR-Ⅱ were decreased significantly in chelidonine high concentration group (P<0.05). CONCLUSIONS: Chelido- nine can inhibit the proliferation, collagen synthesis as well as the protein and mRNA expressions of TβR-Ⅰand TβR-Ⅱ in activated CFSC-8B cells.
期刊: 2019年第30卷第13期
作者: 李晓明,林鹏飞,董妙先,徐天娇,于春磊,荣华,王晓丽
AUTHORS: LI Xiaoming,LIN Pengfei,DONG Miaoxian,XU Tianjiao,YU Chunlei,RONG Hua,WANG Xiaoli
关键字: 白屈菜碱;大鼠肝星状细胞CFSC-8B;细胞增殖;胶原合成;转化生长因子β1受体
KEYWORDS: Chelidonine; Hepatic stellate cells CFSC-8B in rats; Cell proliferation; Collagen synthesis; TGF-β1 receptor
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