1.The effect of LncRNA SNHG14 on high glucose induced podocyte injury by targeting miR-30a-5p
Cuiwen KONG ; Yanshuang LU ; Liping SUN ; Fenfen YU
Tianjin Medical Journal 2025;53(9):903-909
Objective To investigate the effect of long non-coding RNA small nucleolar RNA host gene 14(LncRNA SNHG14)on high glucose-induced podocyte injury by targeting microRNA-30a-5p(miR-30a-5p).Methods Podocytes were cultured in vitro and were divided into the following groups:the standard glucose(NG)group,the high glucose(HG)group,the si-NC+HG group,the si-SNHG14+HG group,the miR-NC+HG group,the miR-30a-5p mimics+HG group,the si-SNHG14+inhibitor NC+HG group and the si-SNHG14+miR-30a-5p inhibitor+HG group.Quantitative real-time polymerase chain reaction(RT-qPCR)was performed to detect expression levels of LncRNA SNHG14 and miR-30a-5p.Flow cytometry was used to determine cell apoptosis.Enzyme-linked immunosorbent assay(ELISA)was applied to measure levels of tumor necrosis factor-alpha(TNF-α),interleukin(IL)-6 and IL-1β.Xanthine oxidase method,ammonium molybdate colorimetry and thiobarbituric acid method were respectively used to detect superoxide dismutase(SOD),catalase(CAT)and malondialdehyde(MDA).Dual-luciferase reporter gene was conducted to verify the targeting relationship between LncRNA SNHG14 and miR-30a-5p.Western blot assay was performed to detect expression levels of apoptosis-related proteins.Results Compared with the NG group,the HG group exhibited increased expression levels of LncRNA SNHG14,cell apoptosis rate,as well as levels of TNF-α,IL-6,IL-1β and MDA,whereas the expression level of miR-30a-5p and levels of SOD and CAT were decreased(P<0.05).Compared with the HG group and the si-NC+HG group,the si-SNHG14+HG group exhibited decreased expression levels of LncRNA SNHG14,apoptosis rate,levels of TNF-α,IL-6,IL-1β and MDA,as well as expression levels of Bax and cleaved caspase-3 proteins,while the expression level of miR-30a-5p,levels of SOD and CAT and the expression level of Bcl-2 protein were increased(P<0.05).Compared with the HG group and the miR-NC+HG group,the miR-30a-5p mimics+HG group showed no significant difference in the expression level of LncRNA SNHG14(P>0.05).Meanwhile,the expression level of miR-30a-5p,levels of SOD and CAT,and expression level of Bcl-2 protein were increased,whereas the cell apoptosis rate,levels of TNF-α,IL-6,IL-1β and MDA,as well as expression levels of Bax and cleaved caspase-3 proteins were decreased(P<0.05).Compared with the si-SNHG14+HG group and the si-SNHG14+inhibitor NC+HG group,the si-SNHG14+miR-30a-5p inhibitor+HG group showed no significant difference in the expression level of LncRNA SNHG14(P>0.05),meanwhile,the expression level of miR-30a-5p,levels of SOD and CAT,and expression level of Bcl-2 protein were reduced,whereas the cell apoptosis rate,levels of TNF-α,IL-6,IL-1β and MDA,as well as expression levels of Bax and cleaved caspase-3 proteins were increased(P<0.05).The dual-luciferase reporter gene assay confirmed that LncRNA SNHG14 targeted and negatively regulated miR-30a-5p.Conclusion The inhibition of LncRNA SNHG14 can target miR-30a-5p to alleviate high glucose induced podocyte injury.
2.The effect of LncRNA SNHG14 on high glucose induced podocyte injury by targeting miR-30a-5p
Cuiwen KONG ; Yanshuang LU ; Liping SUN ; Fenfen YU
Tianjin Medical Journal 2025;53(9):903-909
Objective To investigate the effect of long non-coding RNA small nucleolar RNA host gene 14(LncRNA SNHG14)on high glucose-induced podocyte injury by targeting microRNA-30a-5p(miR-30a-5p).Methods Podocytes were cultured in vitro and were divided into the following groups:the standard glucose(NG)group,the high glucose(HG)group,the si-NC+HG group,the si-SNHG14+HG group,the miR-NC+HG group,the miR-30a-5p mimics+HG group,the si-SNHG14+inhibitor NC+HG group and the si-SNHG14+miR-30a-5p inhibitor+HG group.Quantitative real-time polymerase chain reaction(RT-qPCR)was performed to detect expression levels of LncRNA SNHG14 and miR-30a-5p.Flow cytometry was used to determine cell apoptosis.Enzyme-linked immunosorbent assay(ELISA)was applied to measure levels of tumor necrosis factor-alpha(TNF-α),interleukin(IL)-6 and IL-1β.Xanthine oxidase method,ammonium molybdate colorimetry and thiobarbituric acid method were respectively used to detect superoxide dismutase(SOD),catalase(CAT)and malondialdehyde(MDA).Dual-luciferase reporter gene was conducted to verify the targeting relationship between LncRNA SNHG14 and miR-30a-5p.Western blot assay was performed to detect expression levels of apoptosis-related proteins.Results Compared with the NG group,the HG group exhibited increased expression levels of LncRNA SNHG14,cell apoptosis rate,as well as levels of TNF-α,IL-6,IL-1β and MDA,whereas the expression level of miR-30a-5p and levels of SOD and CAT were decreased(P<0.05).Compared with the HG group and the si-NC+HG group,the si-SNHG14+HG group exhibited decreased expression levels of LncRNA SNHG14,apoptosis rate,levels of TNF-α,IL-6,IL-1β and MDA,as well as expression levels of Bax and cleaved caspase-3 proteins,while the expression level of miR-30a-5p,levels of SOD and CAT and the expression level of Bcl-2 protein were increased(P<0.05).Compared with the HG group and the miR-NC+HG group,the miR-30a-5p mimics+HG group showed no significant difference in the expression level of LncRNA SNHG14(P>0.05).Meanwhile,the expression level of miR-30a-5p,levels of SOD and CAT,and expression level of Bcl-2 protein were increased,whereas the cell apoptosis rate,levels of TNF-α,IL-6,IL-1β and MDA,as well as expression levels of Bax and cleaved caspase-3 proteins were decreased(P<0.05).Compared with the si-SNHG14+HG group and the si-SNHG14+inhibitor NC+HG group,the si-SNHG14+miR-30a-5p inhibitor+HG group showed no significant difference in the expression level of LncRNA SNHG14(P>0.05),meanwhile,the expression level of miR-30a-5p,levels of SOD and CAT,and expression level of Bcl-2 protein were reduced,whereas the cell apoptosis rate,levels of TNF-α,IL-6,IL-1β and MDA,as well as expression levels of Bax and cleaved caspase-3 proteins were increased(P<0.05).The dual-luciferase reporter gene assay confirmed that LncRNA SNHG14 targeted and negatively regulated miR-30a-5p.Conclusion The inhibition of LncRNA SNHG14 can target miR-30a-5p to alleviate high glucose induced podocyte injury.

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