H2-B1基因转染对小鼠内皮细胞凋亡与增殖的影响

目的转染pEGFP-N1-H281质粒载体至小鼠血管内皮细胞（VEC），观察H2-B1基因对小鼠VEC凋亡与增殖的影响，探讨借鉴母胎免疫耐受模型预防心脏移植术后免疫排斥反应的机制。方法0.5μg/ml空质粒、0．5μg／ml H2-B1质粒、1.0μg/ml H2-B1质粒转染小鼠VEC后，采用实时荧光定量PCR、流式细胞仪、酶标仪检测不同时间质粒的转染效率、H2．B1基因mRNA的相对表达量、转染VEC的凋亡与增殖情况。结果荧光定量PCR结果显示，0．5μg/ml、1．0μg／ml H2-B1质粒组的H2-B1基因mRNA表达量均高于对照组（P〈0．05，P〈0．001）。H2-B1基因促进VEC凋亡，转染48h、72h后1．0斗g／mlH2一B1质粒组与空白对照、0．5μg／ml空质粒比较，差异有统计学意义（P〈0．05）。H2-B1基因抑制VEC增殖，转染24h、48h、72h后0．5μg／ml H2-B1质粒组、1．0μg／ml H2-B1质粒组分别与空白对照组、0．5μg／ml空质粒组比较，差异均有统计学意义（P〈0．01）。结论pEGFP—N1-H281质粒载体转染小鼠VEC后，能够有效促进其凋亡，抑制其增殖，减弱小鼠VEC活性，诱导免疫耐受。

Effects of H2-Bl gene transfection on the apoptosis and proliferation of mouse vascular endothelial cell

Objective The pEGFP-N1-H2B1 plasmid vector was transfected to mouse vascular endothelial cell （VEC） to investigate the effects of H2-B1 gene on the apoptosis and proliferation, and then speculated the possible mechanism of immunological rejection after heart transplantation mirroring maternal- fetal immune tolerance. Methods The mouse VEC was transfected by 0.5 μg/ml of pEGFP-N1 plasmid vector, 0.5 μg/ml of pEGFP-N1-H2B1 plasmid vector and 1.0 μg/ml of pEGFP-N1-H2B1 plasmid vector, respectively. The efficiency of transfection was detected; the H2-B1 mRNA level, the apoptosis and proliferation of VEC were also investigated at 24 h ,48 h and 72 h, respectively. Results Expression of the H2-B1 gene was determined by real time quantitative PCR. Expression of the H2-B1 in 0. 5 μg/ml and 1.0 μg/ml of pEGFP-N1-H2B1 plasmid vector groups was higher than that of the control group （ P 〈 0. 05 ,P 〈 0. 001, respectively）. VEC cell apoptosis was promoted after 48 h and 72 h in the 1.0μg/ml H2-B1 gene group compared with control group （ P 〈 0. 05 ）. VEC cell proliferation was inhibited in the H2-B1 gene groups at 24 h, 48 h and 72 h, respectively （ P 〈 0. 01 ）. Conclusions Transfection of pEGFP-N1 -H2B1 plasmid vector to mouse VEC can increase the expression of H2-B1 mRNA, induce the apoptosis and inhibit proliferation, as a result, weaken the cytoactivity and induce immune tolerance.