Role of lncRNA LAMA5-AS1 in insulin resistance and the regulation mechanism of the combination of Astragalus polysaccharide and Berberine
Objective:To investigate whether the combination of Astragalus polysaccharide(AP)and berberine(BBR)can regulate the expression of lncRNA LAMA5-AS1 in HepG2 cell models with improved insulin resistance(IR).Methods:IR HepG2 cell model was established and given APB-BR intervention,overexpression vector and siRNA were constructed,and the overexpression and interference effects were detected.LAMA5-AS1 and LAMA5-AS1 siRNA were transfected into HepG2 cells,and the expression of IR-related genes was detected by qRT-PCR.RNA pull-down and RNA immunoprecipitation(RIP)were used to detect the interaction and binding between LAMA5-AS1 and PCB proteins,respectively.Results:Compared with the control group,LAMA5-AS1 expression was significantly increased in the model group(P＜0.05),and the LAMA5-AS1 expression was significantly decreased after the intervention of AP-BBR(P＜0.05).Compared with IR group,mRNA levels of chemerin,GLUT2,PPAR α and PPAR α in IR+AP-BBR group,IR+OE+AP-BBR group and IR+OENC+AP-BBR group were significantly increased(P＜0.01),chemerin,GLUT2,myostatin,p38MAPK and PPAR α mRNA levels were significantly increased in IR+AP-BBR group,IR+siRNA+AP-BBR group and IR+siNC+AP-BBR group(P＜0.01,P＜0.05);mRNA levels of chemerin,GLUT2,myostatin,p38MAPK,PPAR α and PPARγ were significantly increased in the IR+siRNA group(P＜0.01,P＜0.05).RNA pull-down and RIP experiments showed that the IP group showed an obvious enrichment trend compared with IgG group(P＜0.01),suggesting that LAMA5-AS1-202 could bind to PCB protein.Conclusion:lncRNA LAMA5-AS1 as the upstream gene can induce IR.AP-BBR may improve IR by inhibiting the expression of LAMA5-AS1 and increasing the level of insulin-related genes.
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