N-乙酰半胱氨酸缓解小鼠代谢综合征膀胱纤维化的分子机制  

Molecular mechanism of N-acetylcysteine alleviating bladder fibrosis in mice with metabolic syndrome

作　　者：任亚琳 苏博严 何綦琪[2] REN Ya-lin;SU Bo-yan;HE Qi-qi(College of Animal Sciences and Technology,Guangxi University,Nanning,Guangxi 530004,China;Department of Urology,Lanzhou)

机构地区：[1]广西大学动物科学技术学院,广西南宁530004 [2]兰州大学第二医院泌尿外科,甘肃兰州730030

出　　处：《南方农业学报》2025年第1期314-323,共10页Journal of Southern Agriculture

基　　金：国家自然科学基金项目(81800671,82360156);甘肃省卫生健康行业科研管理项目(GSWSKY2021-070);兰州大学萃英人才面上项目(CY2021-MS-A02)。

摘　　要：【目的】探究N-乙酰半胱氨酸(N-acetylcysteine,NAC)缓解小鼠代谢综合征膀胱纤维化的分子机制,为深入理解膀胱纤维化的病理学过程及开发新的治疗策略提供理论参考。【方法】选择8周龄C57BL/6雄性小鼠和ob/ob(B6.VLepob/J)雄性小鼠(代谢综合征模型),试验持续20周,试验结束后处死小鼠,采集膀胱组织。以C57BL/6雄性小鼠为对照组(Con),ob/ob(B6.V-Lepob/J)雄性小鼠为代谢综合征模型组(ob/ob),利用Western blotting检测小鼠膀胱组织中TGF-β1、SMAD、p38、p-ERK、p-JNK、α-SMA、3-Nitrotyrosine(NT)等蛋白相对表达量,采用免疫组织化学染色检测小鼠膀胱组织中NF-κB与TGF-β1表达情况及分布特点。分离小鼠膀胱平滑肌细胞(BSMCs),筛选葡萄糖浓度并绘制BSMCs生长曲线。设对照组(Con)、高糖组(HG)、高糖+NAC组(HG+NAC)和高渗组(HO),检测CRP、IL-6、IL-1β、NLRP3、TGF-β1、BK-β1、SK_(Ca)3蛋白相对表达量。使用MCC950和PDTC抑制NLRP3和NF-κB信号通路,检测ROCK1、NLRP3、NF-κB、TGF-β1蛋白相对表达量。【结果】与Con组相比,ob/ob组小鼠膀胱组织中NT、TGF-β1和α-SMA蛋白相对表达量显著升高(P<0.05,下同),SMAD、p38、p-ERK和p-JNK蛋白相对表达量无显著差异(P>0.05,下同)。葡萄糖浓度筛选结果表明,45 mmol/L的葡萄糖为最佳干预浓度。Western blotting检测结果显示,与Con组相比,HG组BSMCs中CRP、IL-6、TGF-β1、IL-1β和NLRP3蛋白相对表达量显著升高,BK-β1和SK_(Ca)3蛋白相对表达量显著降低。与HG组相比,HG+NAC组BSMCs中CRP、IL-6、TGF-β1、IL-1β和NLRP3蛋白相对表达量显著降低,BK-β1和SK_(Ca)3蛋白相对表达量显著升高。抑制NLRP3和NF-κB信号通路后,与HG组相比,HG+MCC950和HG+PTCD组BSMCs中TGF-β1蛋白相对表达量显著降低,NF-κB蛋白相对表达量显著升高,HG+MCC950组BSMCs中ROCK1和NLRP3蛋白相对表达量显著降低,HG+PTCD组BSMCs中ROCK1和NLRP3蛋白相对表达量无显著差异。【结论】高糖因【Objective】To explore the molecular mechanism by which N-acetylcysteine(NAC)alleviated bladder fibrosis in metabolic syndrome mice,which could provide theoretical reference for a deeper understanding of the pathological process of bladder fibrosis and the development of new treatment strategies.【Method】C57BL/6 male mice and ob/ob(B6.V-Lepob/J)male mice(metabolic syndrome model)at 8 weeks of age were selected.The experiment lasted for 20 weeks.After the end of the experiment,the mice were killed and the bladder tissues were collected.C57BL/6 male mice were used as the control group(Con),and ob/ob(B6.V-Lepob/J)male mice were used as the metabolic syndrome model group(ob/ob).The relative expression levels of TGF-β1,SMAD,p38,p-ERK,p-JNK,α-SMA,3-Nitrotyrosine(NT)and other proteins in the bladder tissues of mice were detected by Western blotting.Immunohistochemical staining was used to detect the expression and distribution characteristics of NF-κB and TGF-β1 in the bladder tissues of mice.Mouse bladder smooth muscle cells(BSMCs)were isolated,the glucose concentration was screened and the growth curve of BSMCs was drawn.The control group(Con),high glucose group(HG),high glucose+NAC group(HG+NAC)and hyperosmotic group(HO)were set up to detect the relative expression levels of CRP,IL-6,IL-1β,NLRP3,TGF-β1,BK-β1 and SK_(Ca)3 proteins.MCC950 and PDTC were used to inhibit the NLRP3 and NF-κB signaling pathways,and the relative expression levels of ROCK1,NLRP3,NF-κB and TGF-β1 proteins were detected.【Result】Compared with the Con group,the relative expression of NT,TGF-β1 andα-SMA proteins in the bladder tissue of the ob/ob group mice was significantly increased(P<0.05,the same below),and there was no significant difference in the relative expression of SMAD,p38,p-ERK and p-JNK proteins(P>0.05,the same below).The results of glucose concentration screening showed that 45 mmol/L glucose was the optimal intervention concentration.Western blotting results showed that compared with the Con group,the relative

关 键 词：N-乙酰半胱氨酸 代谢综合征 膀胱功能损害 氧化应激 炎症反应 

分 类 号：S865.13[农业科学—野生动物驯养]