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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 |
| TITLE： | |
| 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 |
| 阅读数： | 304 次 |
| 本月下载数： | 2 次 |
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