本文選題：SLIT2-N蛋白　切入點(diǎn)：VEGF165　出處：《重慶醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:探索Slit2-N蛋白對(duì)VEGF165誘導(dǎo)的HUVEC及HMVEC細(xì)胞增殖的影響及其機(jī)制,為尋找治療CNV新的靶點(diǎn)提供思路和線索。方法:1.體外培養(yǎng)HUVEC及HMVEC細(xì)胞株。2.CCK-8篩選Slit2-N蛋白及VEGF165蛋白作用于HUVEC及HMVEV細(xì)胞的有效工作濃度。3.CCK-8檢測(cè)Slit2-N蛋白對(duì)VEGF165誘導(dǎo)的HUVEC及HMVEC細(xì)胞增殖的影響。4.Annexin-V FITC/PI雙染流式法檢測(cè)HUVEC及HMVEC細(xì)胞空白對(duì)照組、Slit2-N組、VEGF165組和Slit2-N and VEGF165組細(xì)胞的凋亡情況。5.CFSE流式法檢測(cè)HUVEC及HMVEC細(xì)胞空白對(duì)照組、Slit2-N組、VEGF165組和Slit2-N and VEGF165組各組細(xì)胞增殖情況。6.Western Blot法檢測(cè)HUVEC及HMVEC細(xì)胞空白對(duì)照組、Slit2-N組、VEGF165組和Slit2-N and VEGF-165組各組細(xì)胞中p-AKT,AKT,p-ERK1/2,ERK1/2表達(dá)情況。7.多組間單因素比較采用單因素方差分析。結(jié)果:1.CCK-8結(jié)果顯示,在48小時(shí)VEGF165能引起HUVEC和HMVEC細(xì)胞明顯增殖的最小濃度分別為25ng/ml和500p M,分別篩選為工作濃度;48小時(shí)Slit2-N不引起HUVEC和HMVEC活力下降的最大濃度均為16n M,篩選為工作濃度。2.細(xì)胞接受處理后48小時(shí)CCK-8結(jié)果提示,Slit2-N組與Control組之間無(wú)顯著差異(HUVEC和HMVEC均p0.05),VEGF165組較Control組明顯上升(HUVEC:P0.01,HMVEC:P0.001),Slit2-N and VEGF165組與VEGF165組之間差異顯著(HUVEC:p0.05,HMVEC:p0.001)。3.Annexin-V FITC/PI雙染流式結(jié)果顯示細(xì)胞早期凋亡率在HUVEC和HMVEC中,24小時(shí),48小時(shí),72小時(shí)VEGF165組與Slit2-N and VEGF165組之間均無(wú)明顯差異(P0.05)。4.在細(xì)胞接受處理后48小時(shí)CFSE染色流式結(jié)果顯示HUVEC細(xì)胞Control組,Slit2-N組,VEGF165組,Slit2-N and VEGF165組中增殖指數(shù)分別為(31.63±0.28),(31.88±0.92),(42.12±1.68),(32.71±0.32);HMVEC各組增殖指數(shù)分別為(12.54±1.08),(13.39±0.15),(15.74±0.51),(12.89±0.90),Slit2-N組與Control組與之間無(wú)明顯差異(HUVEC和HMVEC中均p0.05),VEGF165組與Control組之間有顯著差異(HUVEC:P0.001,HMVEC:P0.01),Slit2-N andVEGF165組與VEGF165組之間有顯著差異(HUVEC:p0.001,HMVEC:p0.01);5.Western Blot結(jié)果顯示,p-AKT的相對(duì)表達(dá)量在Slit2-N組與Control組之間無(wú)顯著差異(HUVEC和HMVEC均p0.05),VEGF165組較Control組明顯上升(HUVEC:p0.001,HMVEC:p0.01),Slit2-N and VEGF165組較VEGF165組明顯下調(diào)(HUVEC:p0.001,HMVEC:p0.01);p-ERK 1/2的相對(duì)表達(dá)量在Slit2-N與Control組之間無(wú)明顯差異(HUVEC和HMVEC均p0.05),VEGF165組較Control組明顯上升(HUVEC:p0.001,HMVEC:p0.001),Slit2-N and VEGF165組與VEGF165組之間無(wú)明顯差異(HUVEC和HMVEC均p0.05)。結(jié)論:Slit2-N可能通過(guò)下調(diào)p-AKT的相對(duì)表達(dá)量,而非p-ERK1/2,抑制VEGF165誘導(dǎo)的HUVEC及HMVEC的細(xì)胞增殖,為尋找治療CNV新的靶點(diǎn)提供了思路和線索。
[Abstract]:Aim: to explore the effect of Slit2-N protein on the proliferation of HUVEC and HMVEC cells induced by VEGF165 and its mechanism.Method 1: 1.Screening the effective working concentration of Slit2-N protein and VEGF165 protein on HUVEC and HMVEV cells in Vitro cultured HUVEC and HMVEC Cell Line .3.The effect of Slit2-N protein on VEGF165 induced proliferation of HUVEC and HMVEC cells. 4. Annexin-V FITC/PI double staining flow cytometry to detect HUVEC and HMVEC cellsThe expression of p-AKTT tachycardia and p-ERK1 / 2 ERK1 / 2 in VEGF165 group and Slit2-N and VEGF-165 group was 7. 7.Single factor analysis of variance (ANOVA) was used to compare single factors among groups.Results 1. The results of CCK-8 showed that the minimum concentration of VEGF165 induced significant proliferation of HUVEC and HMVEC cells at 48 hours was 25ng/ml and 500p / M, respectively, and the maximum concentration of Slit2-N that could not induce the decrease of HUVEC and HMVEC activity was 16nM.In HUVEC and HMVEC, there was no significant difference between VEGF165 group and Slit2-N and VEGF165 group.鍦ㄧ粏鑳?yōu)鎺ュ彈澶勭悊鍚?8灝忔椂CFSE鏌撹壊嫻佸紡緇撴灉鏄劇ずHUVEC緇嗚優(yōu)Control緇，

本文編號(hào)：1719012