解放军医学杂志2018，Vol.43Issue(3)：189-194,6.DOI:10.11855/j.issn.0577-7402.2018.03.02
lnc-MGC对高糖诱导的人腹膜间皮细胞转分化的影响
Effect of lnc-MGC on the trans-differentiation of peritoneal mesothelial cells induced by high glucose
摘要
Abstract
Objective To explore the role of long noncoding RNA-MGC (lnc-MGC) on the trans-differentiation of human peritoneal mesothelial cells (HPMCs) induced by high glucose.Methods The immortalized HPMCs were used to establish control group and high glucose group (60mmol/L) respectively.Cells in control group were cultured with ordinary cell medium,and in high glucose group were stimulated with high glucose medium for 72h.Changes of lnc-MGC expression in the both groups were measured by RT-PCR,and the changes of mRNA and protein expression of E-Cadherin,α smooth muscle actin (α-SMA),connective tissue growth factor (CTGF),type Ⅰ collagen (COL-1) and type Ⅲ collagen (COL-3) in epithelial cells of both groups were measured by RT-PCR and Western blotting.The HPMCs were transfected with lentivirus,and then the changes of the above indexes were observed after up-and down-regulation of lnc-MGC.Results By high glucose stimulation of HPMCs for 72h,RTPCR results showed that the expressions oflnc-MGC,α-SMA,CTGF,COL-1 and COL-3 mRNA increased obviously (P＜0.05),and the expression of E-Cadherin mRNA decreased markedly (P＜0.05) in high glucose group than in control group;Western-blotting results indicated that the expression of protein was consistent with that of mRNA,and the differences were statistically significant (P＜0.05).After lentivirus transfection and down-regulation oflnc-MGC,RT-PCR results showed that the expressions of lnc-MGC,α-SMA,CTGF,COL-1 and COL-3 mRNA decreased obviously (P＜0.05),and the expression of E-Cadherin mRNA increased markedly (P＜0.05) in high glucose group than in control group;Western-blotting results showed that the expression of protein was consistent with that of mRNA,and the differences were statistically significant (P＜0.05).After lentivirus transfection and upregulation of lnc-MGC,RT-PCR results showed that the expressions of lnc-MGC,α-SMA,CTGF,COL-1 and COL-3 mRNA increased significantly (P＜0.05),and the expression of E-Cadherin mRNA decreased obviously (P＜0.05) in high glucose group than in control group;Western blotting results showed that the expression of protein was consistent with that of mRNA,and the differences were statistically significant (P＜0.05).The downstream target was predicted as miRNA126-3p,and compared with the control group,the expression ofmiRNA126-3p increased (P＜0.05) after high glucose stimulation,and after transfection with down regulated lnc-MGC lentivirus,the expression of miRNA126-3p decreased obviously (P＜0.05),and transfection with up regulated lnc-MGC lentivirus,the expression ofmiRNA126-3p increased obviously (P＜0.05).Conclusions lnc-MGC participates in the process of HPMCs transdifferentiation through regulating miRNA126-3p.Regulation oflnc-MGC expression level may control the phenotype transition of HPMCs,and delay the development of peritoneal fibrosis.关键词
长链非编码RNA/腹膜间皮细胞/细胞表型转换Key words
long noncoding RNA/human peritoneal mesothelial cells/cell phenotype transition分类
医药卫生引用本文复制引用
李欢,何丽洁,娄未娟,王汉民..lnc-MGC对高糖诱导的人腹膜间皮细胞转分化的影响[J].解放军医学杂志,2018,43(3):189-194,6.基金项目
This work was supported by the National Natural Science Foundation of China (81770669,81770764,81500581)国家自然科学基金(81770669、81770764、81500581) （81770669,81770764,81500581）
