發(fā)布時(shí)間：2018-08-03 21:46

【摘要】：目的:后發(fā)性白內(nèi)障(posterior capsule opacification,PCO)也稱后囊膜混濁,是白內(nèi)障術(shù)后及晶狀體外傷后最常見(jiàn)的并發(fā)癥。研究表明殘留的晶狀體上皮細(xì)胞增殖在PCO的發(fā)病機(jī)制中發(fā)揮著十分重要的作用,但其增殖的具體原理尚未完全闡明,本研究探討體外應(yīng)用血小板源性生長(zhǎng)因子(platelet derived growth factor,PDGF)及PI3K(phosphatidylinositols-kinase)信號(hào)通路抑制劑LY294002分別或同時(shí)刺激人源晶狀體上皮細(xì)胞(lens epithelial cells,LECs)株SRA01/04,以探討PI3K-AKT信號(hào)通路在PDGF促進(jìn)人晶狀體上皮細(xì)胞增殖中的作用。方法:人晶狀體上皮細(xì)胞株SRA01/04常規(guī)傳代培養(yǎng),用不同濃度的PDGF(0ng/ml,0.1ng/ml,1ng/ml,10ng/ml,100ng/ml),不同濃度LY294002(0μM,5μM,10μM,20μM,40μM)以及不同濃度的PDGF(10ng/ml)+LY294002(0μM,5μM,10μM,20μM,40μM)分別作用于SRA01/04細(xì)胞24小時(shí),MTT法檢測(cè)各組細(xì)胞的增殖率或增殖抑制率;根據(jù)MTT結(jié)果,選定各藥物組濃度,將實(shí)驗(yàn)分為四組,即空白對(duì)照組、PDGF組(PDGF濃度為10ng/ml)、LY294002組(LY294002濃度為40μM)、PDGF+LY294002組(10ng/ml的PDGF+40μM的LY294002),各組細(xì)胞培養(yǎng)24h,倒置顯微鏡下觀察各組晶狀體上皮細(xì)胞增殖、抑制的情況;蛋白印跡法檢測(cè)各組細(xì)胞中總AKT及P-AKT的蛋白表達(dá)量差異。結(jié)果:1、人晶狀體上皮細(xì)胞SRA01/04的增殖率與PDGF的濃度呈正相關(guān)(P0.001);2、人晶狀體上皮細(xì)胞SRA01/04的增殖的抑制率與LY294002的濃度呈正相關(guān)(P0.001);3、PDGF促人晶狀體上皮細(xì)胞SRA01/04的增殖作用與LY294002的濃度呈負(fù)相關(guān)(P0.001);4、PDGF、LY294002分別或同時(shí)干預(yù)人晶狀體上皮細(xì)胞SRA01/04時(shí)細(xì)胞總AKT表達(dá)無(wú)明顯差異(P=0.066);5、PDGF單獨(dú)干預(yù)人晶狀體上皮細(xì)胞SRA01/04時(shí)細(xì)胞p-AKT表達(dá)量增加,差異有統(tǒng)計(jì)學(xué)意義(P0.001);6、LY294002單獨(dú)干預(yù)人晶狀體上皮細(xì)胞SRA01/04時(shí)細(xì)胞p-AKT表達(dá)量減少,差異有統(tǒng)計(jì)學(xué)意義(P0.001);7、PDGF和LY294002同時(shí)干預(yù)人晶狀體上皮細(xì)胞SRA01/04時(shí)細(xì)胞p-AKT表達(dá)量較PDGF單獨(dú)干預(yù)時(shí)減少而較LY294002單獨(dú)干預(yù)時(shí)增加,差異有統(tǒng)計(jì)學(xué)意義(P0.001)。結(jié)論:1、PDGF能促進(jìn)人晶狀體上皮細(xì)胞的增殖;2、LY294002能抑制人晶狀體上皮細(xì)胞的增殖;3、PDGF促進(jìn)人晶狀體上皮細(xì)胞的增殖作用能被LY294002抑制;4、PDGF通過(guò)激活PI3K/AKT信號(hào)通路促進(jìn)人晶狀體上皮細(xì)胞的增殖。
[Abstract]:Objective: posterior capsular opacification (posterior capsule) is the most common complication after cataract surgery and lens trauma. Studies have shown that the residual lens epithelial cell proliferation plays a very important role in the pathogenesis of PCO, but the specific mechanism of its proliferation has not been fully clarified. In this study, we used platelet-derived growth factor (platelet derived growth factor-PDGF (PDGF) and PI3K (phosphatidylinositols-kinase) signal pathway inhibitor LY294002 to stimulate human lens epithelial cell line SRA01 / 04, respectively or simultaneously, in order to explore the effect of PI3K-AKT signaling pathway on the promotion of PDGF on human lens. The role of skin cell proliferation. Methods: human lens epithelial cell line SRA01/04 was subcultured routinely. SRA01/04 cells were treated with different concentrations of PDGF (0 ng / ml 0.1 ng / ml 10 ng / ml 10ng / ml), different concentrations of LY294002 (0 渭 M 5 渭 M 10 渭 M ~ 10 渭 M) and different concentrations of PDGF (10ng/ml) LY294002 (0 渭 M 5 渭 M 5 渭 M 10 渭 M 10 渭 M 10 渭 M 40 渭 M) for 24 hours respectively to detect the proliferation rate or inhibition rate of SRA01/04 cells. Each drug group was selected and divided into four groups: the blank control group (10ng/ml) (LY294002 40 渭 M) and PDGF LY294002 (40 渭 M). The cells were cultured for 24 hours. The proliferation and inhibition of lens epithelial cells in each group were observed under inverted microscope. The protein expression of total AKT and P-AKT was detected by Western blot. Results the proliferation rate of SRA01/04 in human lens epithelial cells was positively correlated with the concentration of PDGF (P0.001) and the inhibitory rate of SRA01/04 proliferation in human lens epithelial cells was positively correlated with the concentration of LY294002 (P0.001). There was no significant difference (P0. 066) in the total AKT expression of human lens epithelial cells (P0. 066). P0. 066 (P0. 066) P0. 05 PDGF alone interfered with the increase of p-AKT expression in SRA01/ 04:00 cells of human lens epithelial cells. The difference was statistically significant (P0.001). LY294002 alone interfered with the decrease of p-AKT expression in SRA01/ 04:00 cells of human lens epithelial cells. The difference was statistically significant (P0. 001). The p-AKT expression of SRA01/ 04:00 cells in human lens epithelial cells treated with P0. 001 PDGF and LY294002 at the same time was lower than that with PDGF alone and increased compared with LY294002 alone. The difference was statistically significant (P0. 001). Conclusion LY294002 can inhibit the proliferation of human lens epithelial cells by activating PI3K/AKT signaling pathway.
【學(xué)位授予單位】：桂林醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R776.1

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