荔枝核总黄酮及罗格列酮对大鼠肝星状细胞PPAR-γ和CTGF表达的影响

发布时间：2018-01-14 13:39

本文关键词：荔枝核总黄酮及罗格列酮对大鼠肝星状细胞PPAR-γ和CTGF表达的影响　出处：《桂林医学院》2016年硕士论文　论文类型：学位论文

【摘要】：目的:研究中药荔枝核总黄酮(total flavonoids of litchi,TFL)对大鼠肝星状细胞(HSC-T6)增殖的影响并与西药罗格列酮作对照,初步探讨TFL抗肝纤维化的相关作用机制。方法:采用筛选出的不同浓度(40、80、160μg/mL)的TFL及15μmol/L的罗格列酮作用于HSC-T6,MTT法检测TFL及罗格列酮对HSC-T6增殖的影响;Quantitative Real-time PCR法检测TFL和罗格列酮对HSC-T6过氧化物酶体增殖物激活受体γ(peroxisome proliferator-activated receptor-γ,PPAR-γ)和结缔组织生长因子(connective tissue growth factor,CTGF)mRNA表达的影响;Westen blot法检测TFL和罗格列酮对HSC-T6 CTGF蛋白表达的影响。结果:TFL作用48、72 h可以明显抑制HSC-T6增殖,且在一定范围(40-160μg/mL)内随时间延长及药物浓度增加,抑制作用更加明显。加药72 h后,高、中、低浓度的TFL及罗格列酮对HSC-T6的抑制率分别为12.53%±0.30%,22.82%±1.24%,30.73%±0.23%,22.47%±0.42%,TFL组A值与正常组比较差异有统计学意义(P0.05),高、低浓度TFL组A值与罗格列酮组比较差异有统计学意义(P0.05),中浓度TFL组A值与罗格列酮组比较差异无统计学意义(P0.05)。TFL和罗格列酮处理72小时后均能上调HSC-T6 PPAR-γmRNA的表达,下调CTGF mRNA及蛋白的表达(p0.05);与罗格列酮组相比,TFL组上调PPAR-γmRNA表达的能力均明显降低(p0.05),低浓度TFL组CTGF mRNA及蛋白表达水平增高(p0.05),中浓度TFL组与罗格列酮组CTGF mRNA及蛋白表达量差异无统计学意义(p0.05),高浓度TFL组CTGF mRNA及蛋白表达水平减低(p0.05);随浓度增加,TFL组PPAR-γmRNA的表达水平依次增高(p0.05),CTGF mRNA及蛋白的表达水平依次减低(p0.05)。结论:TFL和罗格列酮均能够抑制HSC-T6增殖从而拮抗肝纤维化,且在一定范围内有剂量时间依赖性,此种抑制作用可能是通过增强PPAR-γ的活性,从而抑制下游细胞因子CTGF的表达来实现的。
[Abstract]:Objective: to study the total flavonoids of litchi. The effect of TFL on the proliferation of rat hepatic stellate cells (HSC-T6) was compared with rosiglitazone. Objective: to explore the mechanism of anti-hepatic fibrosis by TFL. Methods: different concentrations of TFL were used to study the mechanism of anti-hepatic fibrosis. 160 渭 g / mL TFL and 15 渭 mol/L rosiglitazone were used to detect the effect of TFL and rosiglitazone on HSC-T6 proliferation. Detection of HSC-T6 peroxisome proliferator activated receptor 纬 (纬) by TFL and rosiglitazone by Quantitative Real-time PCR. Peroxisome proliferator-activated receptor- 纬. PPAR- 纬) and connective tissue growth factor (CTGF- 纬) mRNA expression; The effect of TFL and rosiglitazone on the expression of HSC-T6 CTGF protein was detected by Westen blot. The proliferation of HSC-T6 was inhibited obviously at 72 h, and the inhibitory effect was more obvious with the prolongation of time and the increase of drug concentration within a certain range of 40 ~ 160 渭 g / mL. After 72 h of administration, the inhibitory effect was more obvious. The inhibitory rates of high, medium and low concentrations of TFL and rosiglitazone on HSC-T6 were 12.53% 卤0.30% 卤1.2472% 卤1.240.73% 卤0.23%, respectively. The value of A in 22.47% 卤0.42% TFL group was significantly higher than that in the normal group (P 0.05). Compared with rosiglitazone group, the A value of low concentration TFL group was significantly different from that of rosiglitazone group (P 0.05). There was no significant difference in A value between TFL group and rosiglitazone group (P 0.05). Both TFL and rosiglitazone could up-regulate the expression of HSC-T6 PPAR- 纬 mRNA after 72 hours treatment. The expression of CTGF mRNA and protein was down-regulated (P 0.05). Compared with rosiglitazone group, the ability of upregulation of PPAR- 纬 mRNA in TFL group was significantly lower than that in rosiglitazone group (P 0.05). The expression of CTGF mRNA and protein in low concentration TFL group was higher than that in control group (p0.05). There was no significant difference in the expression of CTGF mRNA and protein between TFL group and rosiglitazone group (p 0.05). The expression of CTGF mRNA and protein in high concentration TFL group decreased p0.05; The expression level of PPAR- 纬 mRNA in TFL group increased in turn with the increase of concentration (p0.05). The expression level of CTGF mRNA and protein decreased in turn (P 0.05). Conclusion both CTGF and rosiglitazone can inhibit the proliferation of HSC-T6 and antagonize liver fibrosis. The inhibition may be achieved by increasing the activity of PPAR- 纬 and thus inhibiting the expression of downstream cytokine CTGF.
【学位授予单位】：桂林医学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R259

【参考文献】