Ⅰ型和Ⅴ型胶原蛋白通过YAP核转位调节大鼠胰岛β细胞增殖的研究

YAP nuclear translocation regulates the proliferation of rat’s pancreaticβ-cells cultured on collagenⅠ-orⅤ-coated dishes

目的探讨Yes相关蛋白(YAP)核转位对Ⅰ型和Ⅴ型胶原蛋白包被的大鼠胰岛β细胞(INS-1细胞)增殖的影响。方法将INS-1细胞分为空白组(未包被胶原蛋白,正常培养)、对照组(包被40.00 mg·mL^(-1)的Ⅰ型胶原蛋白)和实验组(包被5.00 mg·mL^(-1)的Ⅴ型胶原蛋白),并培养48 h。用噻唑蓝法检测细胞的存活率,用PI流式细胞术检测细胞周期,用蛋白质印迹法检测YAP蛋白的表达情况。结果实验组、对照组和空白组的干预48 h后细胞存活率分别为(46.65±4.29)%、(112.90±3.20)%和(100.00±0.00)%,S+G2/M期比例分别为(30.51±1.05)%、(41.59±0.90)%和(36.15±1.08)%,细胞核中YAP相对表达水平分别为0.78±0.06、1.17±0.04和1.00±0.00,细胞质中YAP相对表达水平分别为1.15±0.02、0.78±0.05和1.00±0.00,实验组和对照组的上述指标与空白组比较,差异均有统计学意义(均P<0.05)。结论Ⅰ型胶原蛋白包被培养促进YAP核转位,从而促进INS-1细胞增殖;而Ⅴ型胶原蛋白包被培养抑制YAP核转位,从而抑制INS-1细胞增殖。

Objective To study the effect of Yes-associated protein(YAP)nuclear translocation on the proliferation of INS-1 cells cultured on collagenⅠ-orⅤ-coated dishes.Methods The INS-1 cells were divided into blank group(non coated with collagen,normal culture),control group(40.00 mg·mL^(-1) collagenⅠ)and experimental group(5.00 mg·mL^(-1) collagenⅤ),and cultured for 48 h.The cell viabilities were detected by methyl thiazolyl tetrazolium assay.The cell cycle was determined by PI flow cytometry.The expressions of YAP in nuclear and cytoplasm were detected by Western blot.Results The cell survival rates after 48 h intervention of experimental,control and blank groups were(46.65±4.29)%,(112.90±3.20)%and(100.00±0.00)%;S+G2/M phase ratios were(30.51±1.05)%,(41.59±0.90)%and(36.15±1.08)%;the relative expressions of YAP in the nucleus were 0.78±0.06,1.17±0.04 and 1.00±0.00;the relative expressions of YAP in cytoplasm were 1.15±0.02,0.78±0.05 and 1.00±0.00,respectively.Compared with the blank group,the above indexes in the experimental and control groups were statistically significant(all P<0.05).Conclusion CollagenⅠcoated culture promotes the nuclear transcription of YAP,thus promoting the proliferation of INS-1 cells,while the collagenⅤcoated culture inhibits the nuclear translocation of YAP,thereby inhibiting the proliferation of INS-1 cells.