OBJECTIVE:To explore whether fat mass and obesity associated proteins(FTO) is an important target of Qiteng Xiaozhuo granules(QTXZG,芪藤消浊颗粒) medicated serum in regulating proliferation and apoptosis of glomerular...OBJECTIVE:To explore whether fat mass and obesity associated proteins(FTO) is an important target of Qiteng Xiaozhuo granules(QTXZG,芪藤消浊颗粒) medicated serum in regulating proliferation and apoptosis of glomerular mesangial cells.METHODS:Medicated serum was obtained from Sprague-Dawley(SD) rats administered intragastrically with QTXZG decoction.The optimal concentration and intervention time of medicated serum were selected with the cell counting kit 8 assay.Cell proliferation was assessed by 5-ethynyl-2’-deoxyuridine(Ed U) and cell apoptosis was investigated using flow cytometry.The expression of FTO,Proliferating cell nuclear antigen,Cyclin D1,B-cell lymphoma 2(Bcl2) and BCL2 assaciated X was detected by Western blot and Real-time quantitative polymerase chain reaction,respectively.Quantification of the m6A RNA methylation was utilized to determine the total level of m6A methylation modification.RESULTS:Ed U and flow cytometry assays revealed that QTXZG medicated serum can remarkably inhibit proliferation and promote apoptosis of lipopolysaccharide(LPS)-induced human glomerular mesangial cells(HGMCs).The FTO overexpression plasmid could inhibit proliferation and promote apoptosis of LPS-induced HGMCs.The FTO inhibitor(FB23-2) can significantly attenuate the effect of QTZXG medicated serum on inhibiting excessive proliferation and promoting apoptosis.QTXZG medicated serum can significantly increase FTO expression and decrease the level of m6A methylation modification.CONCLUSIONS:FTO is a key target for QTXZG medicated serum in inhibiting excessive proliferation and promoting apoptosis of human glomerular mesangial cells.展开更多
基金National Natural Science Foundation of China:Lnc NONRATG001910.2 Competitively Binds miR-339 to Regulate the Mechanism of Syk Involvement in the Pathogenesis of Chronic Nephritis and the Intervention of Qiteng Xiaozhuo Particles (No.81973546)the Key Scientific Research Projects of Natural Science in Colleges and Universities in Anhui Province:Study on the Mechanism of FTO-Mediated FOXO6 mRNA m6A Modification Regulating the Proliferation and Apoptosis of Glomerular Mesangial Cells (No.2022AH050747)Natural General Projects of Chaohu University:Expression and Target Analysis of miR-155-5p in Kidney Tissue of CGN Rats (No.XLY-201912)
文摘OBJECTIVE:To explore whether fat mass and obesity associated proteins(FTO) is an important target of Qiteng Xiaozhuo granules(QTXZG,芪藤消浊颗粒) medicated serum in regulating proliferation and apoptosis of glomerular mesangial cells.METHODS:Medicated serum was obtained from Sprague-Dawley(SD) rats administered intragastrically with QTXZG decoction.The optimal concentration and intervention time of medicated serum were selected with the cell counting kit 8 assay.Cell proliferation was assessed by 5-ethynyl-2’-deoxyuridine(Ed U) and cell apoptosis was investigated using flow cytometry.The expression of FTO,Proliferating cell nuclear antigen,Cyclin D1,B-cell lymphoma 2(Bcl2) and BCL2 assaciated X was detected by Western blot and Real-time quantitative polymerase chain reaction,respectively.Quantification of the m6A RNA methylation was utilized to determine the total level of m6A methylation modification.RESULTS:Ed U and flow cytometry assays revealed that QTXZG medicated serum can remarkably inhibit proliferation and promote apoptosis of lipopolysaccharide(LPS)-induced human glomerular mesangial cells(HGMCs).The FTO overexpression plasmid could inhibit proliferation and promote apoptosis of LPS-induced HGMCs.The FTO inhibitor(FB23-2) can significantly attenuate the effect of QTZXG medicated serum on inhibiting excessive proliferation and promoting apoptosis.QTXZG medicated serum can significantly increase FTO expression and decrease the level of m6A methylation modification.CONCLUSIONS:FTO is a key target for QTXZG medicated serum in inhibiting excessive proliferation and promoting apoptosis of human glomerular mesangial cells.
