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仙鹤草不同提取部位抗肝纤维化作用的研究
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| 篇名： | 仙鹤草不同提取部位抗肝纤维化作用的研究 |
| TITLE： | Study on the different extraction fractions of Agrimonia pilosa against hepatic fibrosis |
| 摘要： | 目的 研究仙鹤草不同提取部位的抗肝纤维化作用。方法以大鼠肝星状细胞HSC-T6为研究对象，检测不同质量浓度（0.5、5、50、500、5000μg/mL，均以生药量计）的仙鹤草不同提取部位（总提取物、乙酸乙酯部位、石油醚部位和正丁醇部位）对该细胞增殖的影响（作用24、48、72h后），并计算半数抑制浓度（IC50）。采用血小板衍生生长因子（PDGF-BB）刺激HSC-T6细胞以复制肝纤维化细胞模型，采用流式细胞仪检测仙鹤草不同提取部位对HSC-T6细胞凋亡的影响，采用酶联免疫吸附试验检测细胞培养上清液中Ⅰ型胶原蛋白（Col-Ⅰ）表达水平，采用Westernblot法检测细胞中α-平滑肌肌动蛋白（α-SMA）、Col-Ⅰ、B细胞淋巴瘤2（Bcl-2）、Bcl-2相关X蛋白（Bax）、胱天蛋白酶3（caspase-3）的表达水平。结果仙鹤草总提取物、乙酸乙酯部位、石油醚部位和正丁醇部位均可显著升高HSC-T6细胞凋亡率（P＜0.01）；作用24h时，上述各部位的IC50分别为50.17、20.75、5.82、4.09μg/mL。经PDGF-BB干预后，HSC-T6细胞培养上清液中Col-Ⅰ和细胞中Col-Ⅰ、α-SMA、Bcl-2、Bax、caspase-3蛋白表达水平均显著升高（P＜0.01）；经仙鹤草不同提取部位干预后，HSC-T6细胞培养上清液或细胞中上述蛋白表达水平大部分均较PDGF-BB组显著逆转（P＜0.05或P＜0.01），其中仙鹤草正丁醇部位干预作用最为显著。结论仙鹤草不同提取部位均可抑制HSC-T6细胞增殖，并诱导其凋亡；正丁醇部位可能是仙鹤草发挥抗肝纤维化作用的有效部位。 |
| ABSTRACT： | OBJECTIVE To study different extraction fractions of Agrimonia pilosa against h epatic fibrosis. METHODS Using hepatic stellate cells HSC-T 6 of rats as objects ，the effects of different extraction fractions （total extract ，ethyl acetate fraction ， petroleum ether fraction and n-butanol fraction ）with different concentrations （0.5，5，50，500，5 000 μg/mL，calculated by raw drug）of A. pilosa on the proliferation of HSC-T 6 cells were detected （after treated for 24，48，72 h）；median inhibition concentration（IC50）was also caculated. Platelet-derived growth factor （PDGF-BB）was used to induce the activation of HSC-T 6 cells to establish hepatic fibrosis cell model. Flow cytometry was used to detect the effects of different extraction fractions of A. pilosa on apoptosis of HSC-T 6 cells. The expression of collagen Ⅰ（Col-Ⅰ）in the supernatant was detected by enzyme linked immunosorbent assay. The expressions of α-smooth muscle actin （α-SMA），Col-Ⅰ，B-cell lymphoma- 2（Bcl-2），Bcl-2-associated X protein （Bax）and caspase- 3 were detected by Western blot assay. RESULTS Total extract ，ethyl acetate fraction ，petroleum ether fraction and n-butanol fraction of A. pilosa could significantly increase the apoptotic rate of HSC-T 6 cells（P＜0.01）. After treated for 24 h，IC50 of above fractions were 50.17，20.75，5.82，4.09 μg/mL，respectively. After intervened with PDGF-BB ，the expression of Col- Ⅰ in supernatant of HSC-T 6 cells as well as protein expression of Col- Ⅰ，α-SMA，Bcl-2，Bax and caspase- 3 in HSC-T6 cells were increased significantly （P＜0.01）. After intervened with different extraction fractions of A. pilosa ，most of the expressions of above proteins in HSC-T 6 cell culture supernatant or cells were significantly reversed compared with PDGF-BB group （P＜0.05 or P＜0.01）， and the intervention effect of n-butanol fraction of A. pilosa was the most significant. CONCLUSIONS Different extraction fractions of A. pilosa can inhibite the proliferation of HSC-T 6 cells and induce their apoptosis；n-butanol fraction from A. pilosa may be an effective fraction to exert the effect of anti-hepatic fibrosis. |
| 期刊： | 2022年第33卷第03期 |
| 作者： | 李麟，饶春燕，雷青松，黄祎，刘华宝，张长江 |
| AUTHORS： | LI Lin，RAO Chunyan ，LEI Qingsong ，HUANG Yi，LIU Huabao ，ZHANG Changjiang |
| 关键字： | 仙鹤草；肝纤维化；细胞凋亡；有效部位 |
| KEYWORDS： | Agrimonia pilosa ； hepatic fibrosis ； cell |
| 阅读数： | 188 次 |
| 本月下载数： | 11 次 |
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