论文部分内容阅读
目的:探讨泛素蛋白酶体抑制剂MG-132对大鼠肺成纤维细胞凋亡和增殖的影响,并初步探索其机制。方法:18只SD大鼠随机分成模型组(n=9)与对照组(n=9),模型组气管内一次性注入博莱霉素(5mg/kg),对照组气管内注入等量无菌生理盐水,分别于7,28,45d每组处死动物3只,提取成纤维细胞进行培养并用相同浓度的MG-132(1.0μmol/L)对成纤维细胞干预24h,应用MTT比色法检测细胞增殖情况,应用流式细胞仪检测细胞凋亡率,用免疫组化技术半定量观察成纤维细胞α-平滑肌动蛋白(α-SMA)和NF-κB的表达情况。①MG-132对模型组成纤维细胞的抑制和促凋亡的作用强于对照组成纤维细胞(P<0.05),其中,对7d模型组成纤维细胞的促凋亡作用最强;②MG-132干预后,模型组成纤维细胞α-SMA和NF-κB的表达逐渐减少,但仍高于对照组成纤维细胞(P<0.05)。结论:泛素蛋白酶体抑制剂MG-132对肺成纤维细胞有抑制和促凋亡作用,可能与减少肺成纤维细胞向肌成纤维细胞的转化及抑制NF-κB的表达有关。
Objective: To investigate the effect of ubiquitin proteasome inhibitor MG-132 on the apoptosis and proliferation of rat lung fibroblasts and to explore its mechanism. Methods: Eighteen Sprague-Dawley rats were randomly divided into model group (n = 9) and control group (n = 9). Bleomycin (5mg / kg) Then the fibroblasts were extracted and cultured with the same concentration of MG-132 (1.0 μmol / L) for 24 h. MTT assay was used to detect the proliferation of fibroblasts Cell proliferation was detected by flow cytometry. The expression of α-smooth muscle actin (α-SMA) and NF-κB in fibroblasts was semi-quantitatively detected by immunohistochemistry. ① The effect of MG-132 on fibroblast inhibition and apoptosis was stronger than that of control fibroblasts (P <0.05), and the effect of MG-132 on stromal fibroblasts was the strongest; ② After the intervention of MG-132, The expression of α-SMA and NF-κB in model group fibroblasts gradually decreased, but still higher than those in control group (P <0.05). CONCLUSION: Ubiquitin-proteasome inhibitor MG-132 inhibits and promotes apoptosis of lung fibroblasts, which may be related to the reduction of the transformation of lung fibroblasts into myofibroblasts and the inhibition of the expression of NF-κB.