p27在荔枝核总黄酮抑制人肝星状细胞LX2增殖过程中的表达及其意义

发布时间：2018-01-06 02:21

本文关键词：p27在荔枝核总黄酮抑制人肝星状细胞LX2增殖过程中的表达及其意义　出处：《桂林医学院》2015年硕士论文　论文类型：学位论文

【摘要】：目的:通过荔枝核总黄酮(total flavonoids of litchi,TFL)作用于人肝星状细胞LX2,研究TFL对LX2增殖及细胞周期的影响。初步探讨TFL调控LX2细胞周期的相关作用机制,为TFL作为一种新的抗纤维化药物提供理论依据。方法:使用不同浓度(7.8125、15.6250、31.2500、62.5000、125.0000μg/mL)TFL处理LX2。采用CCK8法检测细胞的增殖活力;对细胞进行PI染色后,采用流式细胞仪分析各组细胞周期的分布情况;使用RT-PCR和Western blot技术测定TFL作用后LX2细胞中p27基因mRNA和蛋白的表达。结果:TFL作用48h、72h可抑制LX2细胞增殖,且随着时间延长,抑制效果更加明显。在药物作用72h后,7.8125、15.6250、31.2500、62.5000、125.0000μg/mL的不同浓度TFL对LX2细胞抑制率依次为7.42%±5.66%、9.88%±0.73%、11.55%±3.35%、9.47%±3.67%、9.12%±5.35%,TFL最小剂量加药组与对照组差异无统计学意义(P0.05),而15.6250、31.2500、62.5000、125.0000μg/mL TFL加药组相对各对照组的差异性均具有统计学意义(P0.05)。细胞周期检测结果显示,经TFL干预72h后,7.8125、15.6250μg/mL加药组和对照组细胞周期分布差异不具有统计学意义,而31.2500、62.5000、125.0000μg/mL加药组相较对照组G0/G1期细胞数改变差异无统计学意义(P0.05),S期细胞量明显增多(P0.05),G2/M期细胞量则相应减少,此期改变部分具有差异性,提示TFL可将LX2细胞阻滞在S期。RT-PCR和Western blot技术检测各组p27在RNA和蛋白水平上的表达量,结果提示随着TFL浓度增大,加药组p27的mRNA和蛋白量较对照组明显升高。结论:荔枝核总黄酮抑制人肝星状细胞增殖并将其阻滞在S期,该作用机制可能与p27表达上调相关。
[Abstract]:Objective: to study the effect of total flavonoids of litchion TFL on human hepatic stellate cell (LX2). To study the effect of TFL on the proliferation and cell cycle of LX2 and to explore the mechanism of TFL regulating the cell cycle of LX2. Methods: using different concentrations of TFL as a new antifibrosis drug, 7.8125 (15.6250) and 31.2500 (62.5000) were used. LX2 was treated with 125.0000 渭 g / mL TFL. The proliferative activity of LX2 was detected by CCK8 assay. After Pi staining, the distribution of cell cycle in each group was analyzed by flow cytometry. RT-PCR and Western blot techniques were used to detect the expression of p27 gene mRNA and protein in LX2 cells treated with TFL for 48 h. The proliferation of LX2 cells was inhibited at 72 h, and the inhibitory effect was more obvious with the prolongation of time. The inhibitory rate of 62.5000 渭 g / mL TFL on LX2 cells was 7.42% 卤5.66 and 9.88% 卤0.73% respectively. 11.55% 卤3.35% 卤3.67% 卤9.12% 卤5.35%, there was no significant difference between the control group and the control group (P 0.05). And 15.6250, 31.2500, 62.5000. The difference of 125.000 渭 g / mL TFL addition group was statistically significant compared with the control group (P 0.05). The cell cycle test showed that the cell cycle was treated with TFL for 72 h. There was no significant difference in cell cycle distribution between 7.8125 and 15.6250 渭 g / mL group and control group, but 31.2500 渭 g / mL was 62.5000 渭 g / mL. Compared with the control group, there was no significant difference in the number of cells in the G _ 0 / G _ 1 phase in the 125.000 渭 g / mL group. The number of cells in G _ 2 / M phase decreased correspondingly, and the changes were different in this phase. It was suggested that TFL could block LX2 cells in S phase. RT-PCR and Western blot techniques were used to detect the expression of p27 at RNA and protein levels. The results suggested that with the increase of TFL concentration, the mRNA and protein contents of p27 in the treatment group were significantly higher than those in the control group. Conclusion: the total flavonoids of litchi seeds inhibit the proliferation of human hepatic stellate cells and block them in S phase. This mechanism may be related to the up-regulation of p27 expression.
【学位授予单位】：桂林医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R575.2

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