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目的研究内质网跨膜蛋白IRE1α对人牙周膜成纤维细胞(human periodontal ligament fibroblasts,hPDLFs)增殖的影响。方法将成功构建的人IRE1α基因重组质粒染入hPDLFs细胞,采用免疫印迹法检测IRE1α重组基因在真核细胞内的表达情况,XTT法和流式细胞仪(FCM)检测转染后hPDLFs细胞增殖和细胞周期变化。结果酶切及测序结果证实IRE1α重组质粒构建成功;免疫印迹分析结果证实,IRE1α重组质粒能在hPDLFs细胞内正确表达;在内质网应激(ERS)状态下,与衣霉素(tunicamycin,TM)对照组相比,XTT检测结果显示:IRE1α实验组hPDLFs细胞增殖率明显增高(P<0.01);FCM结果分析显示:IRE1α实验组hPDLFs细胞S期比例增加而G1期减少(P<0.05)。结论在ERS状态下,IRE1α促进hPDLFs细胞增殖,促进hPDLFs细胞从G1期进入S期。
Objective To study the effect of endoplasmic reticulum transmembrane protein IRE1α on the proliferation of human periodontal ligament fibroblasts (hPDLFs). Methods The recombinant plasmid of human IRE1α gene was successfully transfected into hPDLFs cells and the expression of recombinant IRE1α gene in eukaryotic cells was detected by Western blotting. The proliferation and proliferation of hPDLFs were detected by XTT and flow cytometry (FCM) Cell cycle changes. Results The results of enzyme digestion and sequencing confirmed that the constructed recombinant plasmid of IRE1α was successfully constructed. The results of Western blotting showed that the recombinant plasmid of IRE1α was correctly expressed in hPDLFs cells. In the condition of endoplasmic reticulum stress (ERS) ) Compared with the control group. The results of XTT showed that the proliferation rate of hPDLFs was significantly increased in IRE1α experimental group (P <0.01). The results of FCM showed that the proportion of S phase in hPDLFs increased and the G1 phase decreased in IRE1α experimental group (P <0.05). Conclusion In ERS state, IRE1α promotes the proliferation of hPDLFs and promotes the progression of hPDLFs from G1 phase to S phase.