氟非尼酮对大鼠肝星状细胞增殖作用的观察

发布时间：2018-05-21 17:25

本文选题：肝纤维化 + 肝星状细胞　；参考：《中南大学》2012年硕士论文

【摘要】：目的：观察AKF-PD对大鼠肝星状细胞增殖的影响,初步探讨其机制方法： 1.原位灌流法从S-D大鼠肝脏内分离原代大鼠肝星状细胞。 2.分离培养14天的原代肝星状细胞分为：正常组、模型组(10ng/ml PDGF、200μg/ml AKF-PD治疗组(10ng/ml PDGF+200μ g/ml AKF-PD)、400μg/ml AKF-PD治疗组(10ng/ml PDGF+400μg/ml AKF-PD),药物预处理24小时后加入PDGF10ng/ml继续培养24小时,MTT法检测AKF-PD对大鼠肝星状细胞增殖的影响。 3.将分离培养14天的原代肝星状细胞分为正常组、模型组(10ng/ml PDGF)、AKF-PD治疗组(10ng/ml PDGF+400μg/ml AKF-PD)、 PFD治疗组(10ng/ml PDGF+400μg/ml PFD)。药物预处理24小时后加入PDGF10ng/ml继续培养24小时,提取细胞蛋白,Wesern Blot法检测CyclinD1、CyclinE、P27Kip1蛋白表达。结果： 1.与正常组比较,模型组肝星状细胞吸光度值显著增加；与模型组比较,不同浓度的AKF-PD治疗组肝星状细胞吸光度值均明显下降,且400μg/ml AKF-PD治疗组较200μg/ml AKF-PD治疗组下降明显。 2.与正常组比较,模型组肝星状细胞CyclinD1、CyclinE表达明显增加,P27Kip1表达明显降低；与模型组比较,AKF-PD治疗组CyclinD1、CyclinE表达显著降低, P27Kip1表达显著升高。结论： 1. AKF-PD显著抑制大鼠肝星状细胞增殖。 2. AKF-PD可能是通过下调CyclinD1、CyclinE表达和上调P27Kip1表达抑制大鼠肝星状细胞增殖。
[Abstract]:Objective: to observe the effect of AKF-PD on the proliferation of rat hepatic stellate cells and to explore its mechanism. Methods: 1. Primary rat hepatic stellate cells were isolated from S-D rat liver by in situ perfusion. 2. Primary hepatic stellate cells were isolated and cultured for 14 days. The model group was treated with 10ng / ml PDGF200 渭 g/ml AKF-PD. The 10ng / ml PDGF 200 渭 g/ml AKF-PDT 400 渭 g/ml AKF-PD treated group was treated with 10ng / ml PDGF 400 渭 g/ml AKF-PDT. The effect of AKF-PD on the proliferation of rat hepatic stellate cells was detected by MTT assay after 24 hours of pretreatment with PDGF10ng/ml. 3. The primary hepatic stellate cells cultured for 14 days were divided into normal group. The model group was treated with 10ng / ml PDGF400 渭 g/ml AKF-PDD, and the model group was treated with 10ng / ml PDGF 400 渭 g/ml PFD-PDD, and the PFD treated group was 10ng / ml PDGF 400 渭 g/ml PFDD. After pretreatment for 24 hours, PDGF10ng/ml was added to culture for 24 hours. The protein expression of Cyclin D _ (1) and Cyclin E _ (27) Kip1 was detected by Wesern Blot assay. Results: 1. Compared with the normal group, the absorbance of hepatic stellate cells in the model group was significantly increased, and the absorbance value of hepatic stellate cells in the different concentrations of AKF-PD treatment group was significantly lower than that in the 200 渭 g/ml AKF-PD group, and that in the 400 渭 g/ml AKF-PD treatment group was significantly lower than that in the 200 渭 g/ml AKF-PD treatment group. 2. Compared with the normal group, the expression of Cyclin D1 cyclin E in hepatic stellate cells in the model group increased significantly, and the expression of P27Kip1 decreased significantly in the model group, while the expression of Cyclin D1 and P27Kip1 increased significantly in the AKF-PD treatment group compared with the model group. Conclusion: 1. AKF-PD significantly inhibited the proliferation of rat hepatic stellate cells. 2. AKF-PD may inhibit the proliferation of rat hepatic stellate cells by down-regulating the expression of Cyclin E and up-regulating the expression of P27Kip1.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363

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