【摘要】 目的探讨红花黄色素是否通过调控lncRNA FGD5-AS1/miR-302a-3p分子轴而减轻高糖诱导的小鼠足细胞损伤。方法体外培养小鼠足细胞,使用高糖与不同剂量的红花黄色素处理细胞。MTT法检测细胞增殖,流式细胞术检测细胞凋亡,RT-qPCR检测FGD5-AS1、miR-302a-3p表达,双荧光素酶报告实验检测FGD5-AS1、miR-302a-3p的靶向关系。高糖诱导的小鼠足细胞中分别转染pcDNA-FGD5-AS1与si-FGD5-AS1,并检测细胞增殖及凋亡。Western blot法检测cleaved caspase-12、cleaved-PARP、CHOP、p-eIF2α、GRP78的表达。结果高糖处理后细胞存活率与FGD5-AS1的表达降低(P<0.05),细胞凋亡率、cleaved caspase-12、cleaved-PARP、CHOP、p-eIF2α、GRP78蛋白表达及miR-302a-3p的表达升高(P<0.05),红花黄色素可逆转高糖对细胞FGD5-AS1、miR-302a-3p表达及增殖、凋亡、内质网应激的作用,双荧光素酶报告实验证实FGD5-AS1可靶向结合miR-302a-3p,转染pcDNA-FGD5-AS1对高糖诱导的小鼠足细胞增殖、凋亡的作用与红花黄色素的作用相似,转染si-FGD5-AS1可降低红花黄色素对高糖诱导的小鼠足细胞增殖、凋亡的作用。结论红花黄色素可能通过调控FGD5-AS1/miR-302a-3p通路从而减轻高糖诱导的小鼠足细胞损伤。更多还原

【Abstract】 AIM To investigate whether safflower yellow alleviates high glucose induced podocyte injury in mice via regulating the molecular axis of lncRNA FGD5-AS1/miR-302 a-3 p.METHODS Mouse podocytes cultured in vitro were treated with high glucose and different doses of safflower yellow. Subsequently, the mouse podocytes were subjected to determinations of cell proliferation by MTT, apoptosis by flow cytometry, the expressions of FGD5-AS1 and miR-302a-3p by RT-qPCR, and the targeting relationships of FGD5-AS1, miR-302a-3p by dual luciferase report test. High glucose-induced mouse podocytes transfected with pcDNA-FGD5-AS1 and si-FGD5-AS1 respectively, had their cell proliferation and apoptosis detected; and the expressions of cleaved caspase-12, cleaved-PARP, CHOP, p-eIF2α and GRP78 investigated by Western blot. RESULTS High glucose treatment brought forth reduced cell survival rate and the expression of FGD5-AS1(P<0.05); increased cell apoptosis rate, expressions of cleaved caspase-12, cleaved-PARP, CHOP, p-eIF2α, GRP78 proteins and the expression of miR-302a-3p(P<0.05). Safflower yellow counteracted the effect of high glucose on the expressions of FGD5-AS1, miR-302a-3p, proliferation, apoptosis and endoplasmic reticulum stress on cells. The dual luciferase report experiment confirmed the specific FGD5-AS1 target and binding to miR-302a-3p. Unlike the si-FGD5-AS1 transfection, transfection of pcDNA-FGD5-AS1 produced effects similar to that of safflower yellow, in terms of proliferation and apoptosis of high glucose-induced mouse podocytes. CONCLUSION Safflower yellow may alleviate high glucose induced podocyte injury in mice via regulating the FGD5-AS1/miR-302 a-3 p pathway.更多还原