甲基苯丙胺通过miR-129-5p/FAM104B分子轴介导大鼠胚胎成纤维细胞损伤

Methylamphetamine mediates rat embryonic fibroblast injury via miR-129-5p/ FAM104B molecular axis

目的探究甲基苯丙胺(METH)是否通过调控miR-129-5p/FAM104B分子轴从而影响大鼠胚胎成纤维细胞(REF)损伤。方法 MTT实验检测不同浓度甲基苯丙胺对REF细胞活力的影响;流式细胞术检测REF凋亡情况, Western blot检测增殖和凋亡相关蛋白的表达;qRT-PCR检测miR-129-5p与FAM104B的表达表达量;双荧光素酶实验验证miR-129-5p与FAM104B的靶向结合关系。结果甲基苯丙胺可抑制REF细胞增殖,促进细胞凋亡,还可促进miR-129-5p的表达,抑制FAM104B的表达。双荧光素酶实验证实miR-129-5p可靶向调控FAM104B的表达。miR-129-5p过表达明显抑制了细胞增殖,促进细胞凋亡。沉默FMA104B可抑制细胞增殖,促进细胞凋亡。抑制miR-129-5p表达或FMA104B过表达可明显降低METH对REF细胞增殖及凋亡的作用。结论甲基苯丙胺通过调控miR-129-5p/FAM104B分子轴而抑制REF增殖及促进细胞凋亡。

Objective To investigate whether Methylamphetamine(METH) affects rat embryonic fibroblasts by regulating the molecular axis of miR-129-5 p/FAM104 B. Methods MTT assay was used to detect the effect of different concentrations of METH on the viability of REF cells. Flow cytometry was used to detect the apoptosis of REF. Western blot was used to detect the expression of proliferation and apoptosis related proteins. qRT-PCR was used to detect the expression of miR-129-5 p and FAM104 B. Double luciferase experiments verified the targeted binding relationship between miR-129-5 p and FAM104 B. Results METH can inhibit the proliferation of REF cells, promote apoptosis, and also promote the expression of MiR-129-5 p and inhibit the expression of FAM104 B. Double luciferase experiments confirmed that miR-129-5 p can target FAM104 B expression. miR-129-5 p overexpression significantly inhibited cell proliferation and promoted apoptosis. Silencing FMA104 B can inhibit cell proliferation and promote apoptosis. Inhibition of miR-129-5 p expression or FMA104 B overexpression can significantly reduce the effects of METH on REF cell proliferation and apoptosis. Conclusion METH inhibits human embryonic fibroblast proliferation and promotes apoptosis by regulating the molecular axis of miR-129-5 p/FAM104 B.