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作 者:贺绍林 朱菡 HE Shaolin;ZHU Han(Department of Nephrology,the People's Hospital of Dazu,Chongqing 402360,China)
机构地区:[1]重庆市大足区人民医院肾内科,重庆402360
出 处:《医学研究与战创伤救治》2024年第10期1021-1026,共6页Journal of Medical Research & Combat Trauma Care
摘 要:目的探讨长链非编码RNA(LncRNA)肺腺癌转移相关转录子1(MALAT1)对狼疮性肾炎(LN)小鼠肾小球系膜细胞(MC)增殖、凋亡、炎症反应及Notch2/hes1通路的影响。方法取10只MRL/MpJ-Faslpr小鼠为LN组,5只健康MRL/MpJ小鼠为正常组,分离培养LN组小鼠肾小球MC细胞,分为Mock组(正常培养)、Vector组(转染MALAT1过表达空载体pcDNA3.1对照)、MALAT1组(转染MALAT1过表达)、SiNC组(转染敲低SiMALAT1阴性对照SiNC)、SiMALAT1-1/2/3组(转染SiMALAT1-1/2/3)。实时荧光定量PCR(qRT-PCR)检测正常组和LN组小鼠肾组织以及各组MC细胞中MALAT1表达水平;CCK-8和EdU法检测MALAT1对MC细胞增殖的影响;流式细胞术检测各组MC细胞凋亡情况;ELISA检测各组MC细胞中肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)和IL-6水平;Western blot检测MALAT1对MC细胞增殖、凋亡和Notch2/hes1信号通路相关蛋白的影响。结果MALAT1在LN小鼠肾组织中高表达(P<0.05);过表达MALAT1可促进MC细胞增殖和炎症反应,诱导凋亡,同时上调Notch2、hes1蛋白水平;而敲低MALAT1则具有与MALAT1过表达相反的效果(P<0.05)。结论敲低MALAT1可抑制LN小鼠肾小球MC细胞增殖和炎症反应,诱导凋亡,其作用机制可能与抑制Notch2/hes1信号通路有关。Objective To investigate the effects of long non-coding RNA(LncRNA)metastasis-associated transcripton-1(MALAT1)on the proliferation,apoptosis,inflammatory response and Notch2/hes1 pathway of mesangial cells(MC)in mice with lupus nephritis(LN).Methods Ten MRL/MPJ-FASLPR mice were selected as LN group and five healthy MRL/MpJ mice were selected as normal group.They were divided into Mock group(normal culture),Vector group(control after transfection MALAT1 overexpression empty vector pcDNA3.1),MALAT1 group(transfection MALAT1 overexpression),SiNC group(transfection with low Si‐MALAT1 negative control SiNC),SIMalat1-1/2/3 groups(transfection SiMAL AT1-1/2/3).Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect MALAT1 expression in kidney tissue and MC cells of mice in normal and LN groups.CCK-8 and EdU methods was used to detect the effect of MALAT1 on the proliferation of MC cells.Flow cytometry was used to detect the apoptosis of MC cells in each group.ELISA was used to detect the levels of tumor necrosis factorα(TNF-α),interleukin-1β(IL-1β)and IL-6 in MC cells.Western Blot was used to detect the effects of MALAT1 on proliferation,apoptosis and proteins associated with Notch2/hes1 signaling pathway of MC cells.Results MALAT1 was highly expressed in kidney tissue of LN mice(P<0.05).Overexpression of MALAT1 could promote the proliferation and inflammation of MC cells,induce apoptosis,and up-regulate the protein levels of Notch2 and hes1.MALAT1 knockdown had the opposite effect to MALAT1 overexpression(P<0.05).Conclusion MALAT1 knockdown can inhibit the proliferation and inflammatory response of MC cells in LN mice and induce apoptosis.The mechanism may be related to the inhibition of Notch2/hes1 signaling pathway.
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