lncRNA ZFAS1调节miR-136-5p/IGF1R轴对多囊卵巢综合征小鼠胰岛素抵抗的影响

Influence of lncRNA ZFAS1 on insulin resistance in mice with polycystic ovary syndrome by regulating mi R-136-5p/IGF1R axis

目的探究长链非编码RNA(lncRNA)锌指结构反义转录本1(ZFAS1)调节miR-136-5p/胰岛素样生长因子1受体(IGF1R)轴对多囊卵巢综合征(PCOS)小鼠胰岛素抵抗(IR)的影响。方法随机把C57BL/6J小鼠分为Ctrl组、PCOS组、Si-NC组、Si-ZFAS1组、antagomir-NC组、miR-136-5p antagomir组、miR-136-5p antagomir+Si-ZFAS1组,采用高脂饮食联合脱氢表雄酮建立PCOS小鼠模型(Ctrl组除外),检测性激素指标、糖代谢指标水平变化;H-E染色观察卵巢组织形态学特征;RT-qPCR检测卵巢组织ZFAS1、miR-136-5p表达水平;Western blot检测卵巢组织IGF1R蛋白表达水平;双荧光素酶报告基因法(DLR)验证ZFAS1/miR-136-5p、miR-136-5p/IGF1R靶向关系。结果与Ctrl组比较,PCOS组小鼠性激素水平紊乱,卵巢呈多囊样改变,伴随明显胰岛素抵抗,同时卵巢组织中ZFAS1、IGF1R表达增加,miR-136-5p表达减少;下调ZFAS1表达后,PCOS小鼠上述症状明显改善,同时卵巢组织中miR-136-5p表达增加,IGF1R表达减少;下调miR-136-5p表达后,PCOS小鼠上述症状明显加重,同时卵巢组织中IGF1R表达增加;在下调ZFAS1表达的同时亦下调miR-136-5p表达,下调ZFAS1表达对PCOS小鼠的改善作用被削弱。DLR检测结果显示,ZFAS1与miR-136-5p,miR-136-5p与IGF1R存在靶向作用关系。结论ZFAS1在PCOS小鼠内呈高表达,抑制其表达可减轻PCOS小鼠卵巢多囊样改变及胰岛素抵抗,可能通过调节miR-136-5p/IGF1R轴而实现。

Objective To explore the influence of long non-coding RNA(lncRNA)zinc finger antisense 1(ZFAS1)on insulin resistance(IR)in polycystic ovary syndrome(PCOS)mice by regulating the miR-136-5p/insulin-like growth factor 1 receptor(IGF1R)axis.Methods C57BL/6J mice were randomly grouped into Ctrl group,PCOS group,Si-NC group,Si-ZFAS1 group,antagomir-NC group,miR-136-5p antagomir group,and miR-136-5p antagomir+Si-ZFAS1 group,high-fat diet combined with dehydroepiandrosterone was used to establish PCOS mouse model(except Ctrl group),and changes in the levels of sex hormone indicators,glucose metabolic indicators were measured.H-E staining was performed to observe the morphological characteristics of ovarian tissue.RT-qPCR was performed to detect the expression levels of ZFAS1 and miR-136-5p in ovarian tissue.Western blot was performed to detect the expression level of IGF1R protein in ovarian tissue;dual-luciferase reporter assay(DLR)was performed to validate the targeting relationship between ZFAS1/miR-136-5p and miR-136-5p/IGF1R.Results Compared with the Ctrl group,PCOS group showed abnormal sex hormone levels,polycystic ovary changes,and obvious insulin resistance,accompanied by increased expression of ZFAS1 and IGF1R in ovarian tissues,while decreased expression of miR-136-5p.After down-regulating the expression of ZFAS1,the above symptoms were significantly improved in PCOS mice,while the expression of miR-136-5p was increased and the expression of IGF1R was decreased in ovarian tissues.After down-regulating the expression of miR-136-5p,the above symptoms were significantly aggravated in PCOS mice,while the expression of IGF1R in ovarian tissues was increased.The expression of miR-136-5p was also down-regulated while the expression of ZFAS1 was down-regulated,and the improvement effect of down-regulated ZFAS1 expression on PCOS mice was weakened.The results of DLR detection showed that there was a targeting relationship between ZFAS1 with miR-136-5p,and between miR-136-5p with IGF1R.Conclusion ZFAS1 is highly expressed in PCOS mice,and inhibiting its expression can alleviate polycystic ovary changes and insulin resistance in PCOS mice,which may be achieved by regulating the miR-136-5p/IGF1R axis.