本文關鍵詞：p27在荔枝核總黃酮抑制人肝星狀細胞LX2增殖過程中的表達及其意義　出處：《桂林醫(yī)學院》2015年碩士論文　論文類型：學位論文

  更多相關文章： 荔枝核總黃酮 人肝星狀細胞 增殖 S期 p27

【摘要】：目的:通過荔枝核總黃酮(total flavonoids of litchi,TFL)作用于人肝星狀細胞LX2,研究TFL對LX2增殖及細胞周期的影響。初步探討TFL調控LX2細胞周期的相關作用機制,為TFL作為一種新的抗纖維化藥物提供理論依據(jù)。方法:使用不同濃度(7.8125、15.6250、31.2500、62.5000、125.0000μg/mL)TFL處理LX2。采用CCK8法檢測細胞的增殖活力;對細胞進行PI染色后,采用流式細胞儀分析各組細胞周期的分布情況;使用RT-PCR和Western blot技術測定TFL作用后LX2細胞中p27基因mRNA和蛋白的表達。結果:TFL作用48h、72h可抑制LX2細胞增殖,且隨著時間延長,抑制效果更加明顯。在藥物作用72h后,7.8125、15.6250、31.2500、62.5000、125.0000μg/mL的不同濃度TFL對LX2細胞抑制率依次為7.42%±5.66%、9.88%±0.73%、11.55%±3.35%、9.47%±3.67%、9.12%±5.35%,TFL最小劑量加藥組與對照組差異無統(tǒng)計學意義(P0.05),而15.6250、31.2500、62.5000、125.0000μg/mL TFL加藥組相對各對照組的差異性均具有統(tǒng)計學意義(P0.05)。細胞周期檢測結果顯示,經(jīng)TFL干預72h后,7.8125、15.6250μg/mL加藥組和對照組細胞周期分布差異不具有統(tǒng)計學意義,而31.2500、62.5000、125.0000μg/mL加藥組相較對照組G0/G1期細胞數(shù)改變差異無統(tǒng)計學意義(P0.05),S期細胞量明顯增多(P0.05),G2/M期細胞量則相應減少,此期改變部分具有差異性,提示TFL可將LX2細胞阻滯在S期。RT-PCR和Western blot技術檢測各組p27在RNA和蛋白水平上的表達量,結果提示隨著TFL濃度增大,加藥組p27的mRNA和蛋白量較對照組明顯升高。結論:荔枝核總黃酮抑制人肝星狀細胞增殖并將其阻滯在S期,該作用機制可能與p27表達上調相關。
[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% 鹵5.66 and 9.88% 鹵0.73% respectively. 11.55% 鹵3.35% 鹵3.67% 鹵9.12% 鹵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.
【學位授予單位】：桂林醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R575.2

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