摘要
目的:研究微小RNA-27a-3p(miR-27a-3p)对糖尿病患者创面愈合的影响及作用机制。方法:采用qPCR和Western blot检测糖尿病患者创面愈合组织中miR-27a-3p、同源异型盒基因A5(HOXA5)mRNA和HOXA5蛋白的表达。使用高糖处理人微血管内皮细胞(HMECs),模拟糖尿病HMECs的损伤。在HMECs中转染miR-27a-3p mimic或HOXA5 siRNA,并进行高糖处理,MTT法测定细胞活力,Transwell小室法检测细胞迁移和侵袭能力,Western blot法测定HOXA5、细胞周期蛋白D1(cyclin D1)、基质金属蛋白酶2(MMP2)、基质金属蛋白酶9(MMP9)和β-连环蛋白(β-catenin)蛋白的表达。StarBase预测结合双萤光素酶活性检测分析miR-27a-3p和HOXA5的靶向关系。在HMECs中共转染miR-27a-3p mimic和pcDNA-HOXA5,并使用高糖处理,观察其对细胞活力、迁移和侵袭能力的影响。结果:糖尿病患者创面愈合组织中miR-27a-3p的表达量明显减少,HOXA5的mRNA和蛋白表达量显著增加(P<0.05)。高糖诱导的HMECs中miR-27a-3p的表达量、细胞活力、细胞迁移和侵袭数量及cyclin D1、MMP2和MMP9的蛋白水平均显著降低(P<0.05)。高表达miR-27a-3p或低表达HOXA5明显增加高糖处理HMECs的细胞活力、迁移和侵袭能力及cyclin D1、MMP2和MMP9的蛋白表达量(P<0.05)。miR-27a-3p靶向调控HOXA5表达。高表达HOXA5可以部分消除miR-27a-3p mimic对高糖处理的HMECs活力、迁移和侵袭能力以及cyclin D1、MMP2、MMP9和β-catenin蛋白表达的影响。结论:miR-27a-3p靶向HOXA5基因,并通过Wnt/β-catenin信号通路促进高糖诱导的微血管内皮细胞生长、迁移和侵袭,从而促进糖尿病患者的创面愈合。
AIM:To study the effect of microRNA-27a-3b(miR-27a-3p)on healing of diabetic wounds.METHODS:The expression of miR-27a-3p,Homeobox A5(HOXA5)at mRNA and protein levels in wound healing tissues of diabetic patients were determined by qPCR and Western blot.Human microvascular endothelial cells(HMECs)were treated with high glucose to simulate the injury of diabetic HMECs.HMECs were transfected with miR-27a-3p mimic or HOXA5 siRNA,and then treated with high glucose.The cell viability was measured by MTT assay.The migration and invasion abilities were detected by Transwell chamber method.The protein expression of HOXA5,cyclin D1,matrix metalloproteinase(MMP)2,MMP9 andβ-catenin was determined by Western blot.StarBase prediction combined with the detection of dual luciferase activity was conducted to analyze the targeting relationship of miR-27a-3p and HOXA5.miR-27a-3p mimic and pcDNA-HOXA5 were co-transfected into HMECs,and the cells were treated with high glucose to observe their effects on cell viability,migration and invasion.RESULTS:The expression of miR-27a-3p was significantly reduced,and the expression of HOXA5 at mRNA and protein levels was dramatically increased in diabetic wound healing tissues(P<0.05).The expression of miR-27a-3p,cell viability,number of migratory cells,number of invasive cells,and the protein levels of cyclin D1,MMP2 and MMP9 in HMECs induced by high glucose were statistically reduced(P<0.05).High expression of miR-27a-3p or low expression of HOXA5 obviously increased the cell viability,the number of migratory cells,the number of invasive cells,and the protein expression levels of cyclin D1,MMP2 and MMP9 in high glucose-treated HMECs(P<0.05).miR-27a-3p targeted HOXA5 expression.High expression of HOXA5 partially reversed the effect of miR-27a-3p mimic on the viability,migration and invasion abilities,and the protein expression of cyclin D1,MMP2,MMP9 andβ-catenin in high glucose-treated HMECs.CONCLUSION:miR-27a-3p targets HOXA5 gene through Wnt/β-catenin signaling pathway,and promotes the v
作者
葛嘉媛
温立霞
李勤
GE Jia-yuan;WEN Li-xia;LI Qin(Guangzhou University of Chinese Medicine,Guangzhou 510000,China;The First People's Hospital of Yichang,Yichang 443000,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2021年第2期292-299,共8页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81571910)。
