异鼠李素通过抑制LncRNA-gm33782减轻AKI肾小管炎性细胞凋亡OA北大核心CSTPCD
Isorhamnetin alleviates acute kidney injury renal tubular inflammatory cell apoptosis by inhibiting LncRNA-gm33782
目的 探讨长链非编码RNA-gm33782(LncRNA-gm33782)在急性肾损伤(acute kidney injury,AKI)肾小管损伤中的功能以及异鼠李素改善肾小管炎性细胞凋亡的作用机制.方法 将 48 只雄性C57BL/6J小鼠随机分为对照组、模型组、治疗组(AKI 模型构建后灌胃给予 30 mg/kg 异鼠李素)、空载质粒电转染组、LncRNA-gm33782 敲低组、LncRNA-gm33782 过表达组、LncRNA-gm33782 敲低组+模型组、LncRNA-gm33782 过表达+治疗组 8 组,通过腹腔一次性注射 20 mg/kg顺铂诱导小鼠AKI,原位电转染技术用于介导肾LncRNA-gm33782 的敲低和过表达.采用Chirp实验捕获AKI肾LncRNA-gm33782 结合蛋白进行质谱分析,挖掘LncRNA-gm33782 直接作用靶蛋白;通过观察电转染敲低LncRNA-gm33782 和异鼠李素(isorhamnetin,ISO)治疗干预后小鼠肾功能、病理结构改变、肾炎性因子(IL-1β、IL-6、TNF-α)表达,评价LncRNA-gm33782 在AKI中的作用;NF-κB被作为介导炎症的关键信号通路,Bax、Bcl-2蛋白表达量以及流式凋亡检测结果被用于评价异鼠李素对AKI肾小管炎性细胞凋亡的治疗作用.结果 AKI小鼠肾表现出严重的肾小管损伤以及巨噬细胞浸润和炎症,异鼠李素的治疗干预和LncRNA-gm33782 电转染敲低均能够减轻AKI肾损伤.LncRNA-gm33782 主要表达于AKI损伤的肾小管细胞,质谱检测发现补体因子-H(complement factor-H,CFH)与其有直接结合关系.体外过表达LncRNA-gm33782 后,CFH表达量随即升高,而异鼠李素对AKI细胞炎性凋亡的治疗作用受到抑制.结论 异鼠李素是通过抑制LncRNA-gm33782 对CFH的调控作用减轻AKI肾小管细胞炎性凋亡.
Objective To investigate the role of LncRNA-gm33782 in tubular injury in acute kidney injury(AKI)and the mechanism by which isorhamnetin ameliorates inflammatory cell apoptosis of tubular cells in AKI.Methods Forty-eight male C57BL/6J mice were randomly assigned to four groups:control group,AKI model group,electroporation+AKI group,and treatment group(oral administration of 30 mg/kg isorhamnetin).AKI was induced by a one-time intraperitoneal injection of 20 mg/kg cisplatin.Chromatin isolation by RNA purification was used to capture the LncRNA-gm33782 binding protein in AKI-affected kidneys for mass spectrometry,revealing the direct target protein of LncRNA-gm33782.The role of LncRNA-gm33782 in AKI was evaluated by observing the renal function,pathological structure,and expression of renal inflammatory factors(interleukin-1β,interleukin-6,and tumor necrosis factor-α)in mice after electrotransfection and isorhamnetin treatment.Nuclear factor-κB was detected as a critical mediator of inflammation.The expression levels of Bax and Bcl-2 protein were detected and flow cytometry was performed to evaluate the therapeutic effect of isorhamnetin on tubular cell apoptosis in AKI.Results Kidneys with cisplatin-induced AKI showed severe renal tubule injury,macrophage infiltration,and inflammation.Isorhamnetin treatment and LncRNA-gm33782 electrotransfection knockdown alleviated these signs of AKI.LncRNA-gm33782 was mainly expressed in renal tubular cells with AKI.LncRNA-gm33782 binding protein was detected by mass spectrometry,and complement factor H was found to have a direct binding relationship with LncRNA-gm33782.After LncRNA-gm33782 was overexpressed in vitro,the expression of complement factor-H immediately increased,and the therapeutic effect of isorhamnetin on apoptosis of AKI-affected inflammatory cells was inhibited.Conclusions Isorhamnetin alleviates apoptosis of tubular inflammatory cells in AKI by inhibiting the regulatory effect of LncRNA-gm33782 on complement factor H.
贾建;谭睿陟;钟霞;粟宏伟;王丽
西南医科大学附属中医医院中西医结合研究中心,四川 泸州 646000西南医科大学附属中医医院泌尿外科,四川 泸州 646000
生物学
急性肾损伤LncRNA-gm33782肾原位电转染异鼠李素炎性凋亡
acute kidney injuryLncRNA-gm33782renal in situ electroporationisorhamnetininflammatory cell apoptosis
《中国实验动物学报》 2024 (006)
762-771 / 10
西南医科大学应用基础项目(2021ZKQN120),西南医科大学(2023ZYYQ08),泸州市科技局创新苗子项目(2021-RCM-118),四川省科技厅项目(2022YFS0621).Funded by Application Foundation Project of Southwest Medical University(2021ZKQN120),Southwest Medical University(2023ZYYQ08),Luzhou Science and Technology Bureau Innovative Seedling Project(2021-RCM-118),Project of Science and Technology Department of Sichuan Province(2022YFS0621).
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