Effect and mechanism of silencing lncRNA XIST on dilated cardiomyopathy induced by adriamycin
Objective To investigate the effect and mechanism of long non-coding RNA X-inactive specific transcript(lncRNA XIST)on myocardial fibrosis and apoptosis in doxorubicin-induced dilated cardiomyopathy(DCM)rats.Methods The DCM rat model was established by adriamycin induction method.The DCM rats were divided into model group,sh-NC group and sh-XIST group,with 10 rats in each group,and another 10 healthy SD rats were selected as control group.Left ventricular end-systolic diameter(LVESD),left ventricular end-diastolic diameter(LVEDD)and left ventricular ejection fraction(LVEF)were measured by ultrasound imaging system.The histopathological changes of myocardium were detected by HE staining and Masson trichromatic staining,and the apoptosis of myo-cardium was detected by TUNEL.The H9c2 cells were treated with adriamycin to induce DCM cell model.DCM cells were divided into model group,sh-NC group,sh-XIST group,sh-XIST+miR-NC group and sh-XIST+miR-149-5p inhibitor group,and H9c2 cells without adriamycin treatment were treated as control group.The targeting relationships between XIST,miR-149-5p and PDGFC were analyzed by dual luciferase reporting assay.The apoptosis level of cardiomyocytes was detected by flow cytometry.The expression levels of XIST,miR-149-5p and PDGFC in rat myocardial tissue and H9c2 cells were detected by RT-qPCR.The expression levels of PDGFC,α-SMA,collagen Ⅰ and TGF-β1 in rat myocardial tissue and H9c2 cells were detected by Western blot.Results ①Compared with sh-NC group,the levels of XIST,and PDGFC mRNA and protein in myocardial tissue of rats were decreased in sh-XIST group(P＜0.01),LVESD,LVEDD,TUNEL positive cell rate,and the protein levels of α-SMA,collagen Ⅰ and TGF-β1 were decreased(P＜0.05),while the level of miR-149-5p and LVEF were increased(P＜0.01).Compared with sh-NC group,both myocardial cell necrosis and tissue fibrosis were reduced in sh-XIST group.②Compared with sh-NC group,the apoptosis rate of H9c2 cells,and the levels of XIST,PDGFC mRNA and protein,α-SMA,collagen Ⅰ and TGF-β1 protein in H9c2 cells were decreased in sh-XIST group(P＜0.05),while the level of miR-149-5p was increased(P＜0.05).Compared with sh-XIST+miR-NC group,the apoptosis rate of H9c2 cells,and the levels of PDGFC mRNA and protein,α-SMA,collagen Ⅰ and TGF-p1 proteins in H9c2 cells were increased in sh-XIST+miR-149-5p inhibitor group(P＜0.05),while the level of miR-149-5p was decreased(P＜0.05).Conclusion Silencing lncRNA XIST can inhibit the adriamycin-induced myocardial fibrosis and the apoptosis in DCM rats by upregulating the expression of miR-149-5p and inhibiting the expression of PDGFC,thereby improving the development of DCM.
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维普
