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目的探讨骨形态发生蛋白-7(BMP-7)修饰的自体骨髓间充质干细胞(BM-MSCs)移植对缺血再灌注损伤(IRI)后肾小管上皮细胞增殖和凋亡的影响。方法 20只新西兰大白兔,兔龄6~8周,体质量(1.5±0.3)kg。采用随机数字表的方法分为假手术组(Ⅰ组)、对照组(Ⅱ组)、单纯BM-MSCs移植组(Ⅲ组)和BMP-7修饰的BM-MSCs移植组(Ⅳ组)(n=5)。构建BMP-7重组腺病毒并转染兔BM-MSCs,建立肾脏IRI模型后,移植组(Ⅲ组和Ⅳ组)于肾血流恢复后经肾动脉推注单纯和基因修饰的自体BM-MSCs,Ⅱ组同法推注等量0.9%氯化钠溶液。4组于术后7 d取兔肾组织,采用TUNEL法检测细胞凋亡,增殖细胞核抗原(PCNA)免疫组织化学检测细胞增殖,分别计算细胞凋亡指数(AI)和增殖指数(PI);免疫组织化学法检测Bcl-2和Bax的表达并计算Bcl-2/Bax比值。结果Ⅳ组AI明显低于Ⅱ组和Ⅲ组(8.38±1.91 vs 29.82±5.19、19.11±3.41,P<0.05),PI明显高于Ⅱ组和Ⅲ组(40.38±7.09 vs 9.89±2.15、24.83±5.88,P<0.05);Ⅳ组Bax蛋白表达最低,Bcl-2蛋白表达及Bcl-2/Bax比值最高(P<0.05)。结论 BMP-7修饰的BM-MSCs移植可以抑制IRI后肾小管上皮细胞凋亡并促进其增殖,从而有利于肾小管损伤的早期修复。
Objective To investigate the effects of BMP-7 modified bone marrow mesenchymal stem cells (BM-MSCs) transplantation on the proliferation and apoptosis of renal tubular epithelial cells after ischemia-reperfusion injury (IRI). Methods Twenty New Zealand rabbits were rabbits aged 6-8 weeks with a body weight of (1.5 ± 0.3) kg. The patients were randomly divided into sham-operation group (groupⅠ), control group (groupⅡ), BM-MSCs transplantation group (groupⅢ) and BM-MSCs transplantation group (groupⅣ) = 5). After the establishment of renal IRI model, the BMP-7 recombinant adenovirus was transfected into rabbit BM-MSCs and the renal allograft BM-MSCs , The same method group Ⅱ bolus 0.9% sodium chloride solution. Rabbits were sacrificed on the 7th day after operation. Cell apoptosis was detected by TUNEL method. Proliferation cell nuclear antigen (PCNA) was detected by immunohistochemistry. Cell apoptosis index (AI) and proliferation index (PI) were calculated. Immunization Histochemistry was used to detect the expression of Bcl-2 and Bax and to calculate the Bcl-2 / Bax ratio. Results AI in group Ⅳ was significantly lower than that in group Ⅱ and group Ⅲ (8.38 ± 1.91 vs 29.82 ± 5.19, 19.11 ± 3.41, P <0.05), and PI was significantly higher than those in group Ⅱ and Ⅲ (40.38 ± 7.09 vs 9.89 ± 2.15 and 24.83 ± 5.88, P <0.05). The expression of Bax, Bcl-2 and Bcl-2 / Bax were the highest in group Ⅳ (P <0.05). Conclusion BMP-7-modified BM-MSCs transplantation can inhibit renal tubular epithelial cell apoptosis and promote its proliferation after IRI, which is beneficial to the early repair of renal tubular injury.