BACKGROUND Hepatic stellate cell(HSC)activation is key to liver fibrosis.Targeting DNA methylation shows promise.Zebularine,a methylation inhibitor,may suppress HSC activation via the calcineurin(CaN)/NFAT3 pathway.Ma...BACKGROUND Hepatic stellate cell(HSC)activation is key to liver fibrosis.Targeting DNA methylation shows promise.Zebularine,a methylation inhibitor,may suppress HSC activation via the calcineurin(CaN)/NFAT3 pathway.Magnetic resonance imaging(MRI)is a noninvasive tool for evaluating liver fibrosis evaluation tool,but multiparametric MRI for zebularine’s effects in liver fibrosis mouse models has not been studied.AIM To clarify the anti-fibrosis mechanism and MRI-evaluated efficacy of zebularine.METHODS In vitro,transforming growth factor(TGF)-β1-stimulated human HSCs(LX-2)were treated with zebularine.α-smooth muscle actin,fibrotic and anti-fibrotic gene levels,and regulator of calcineurin1(RCAN1)regulation were measured.In vivo,carbon tetrachloride(CCl_(4))-induced liver fibrosis in mice was treated with zebularine,and fibrosis was evaluated using various biochemical,histopathological,and MRI methods.RESULTS Zebularine upregulated RCAN1.4 protein(P<0.01)and inhibited the CaN/NFAT3 pathway(P<0.05).In HSCs,TGF-β1 reduced anti-fibrotic gene massage RNA(mRNA)and increased fibrotic mRNA(P<0.05),whereas zebularine had the opposite effects(P<0.01,P<0.05).CCl4-treated mice exhibited increases in various fibrosis-related indices,all of which were reversed by zebularine treatment(P<0.05).CONCLUSION Zebularine may reduce LX-2 activation and extracellular matrix deposition via RCAN1.4 and CaN/NFAT3 path-ways.Multiparametric MRI can assess its efficacy,suggesting zebularine’s potential as a liver fibrosis treatment.展开更多
目的:探究游泳和下坡跑通过钙调磷酸酶(CN)/活化T细胞核因子(NFAT)途径对T2DM小鼠骨吸收代谢的影响。方法:采用6周高脂膳食和一次性注射链脲佐菌素(STZ)进行T2DM造模,成功后随机分为T2DM对照组(TC)、T2DM游泳组(TS)和T2DM下坡跑组(TD),...目的:探究游泳和下坡跑通过钙调磷酸酶(CN)/活化T细胞核因子(NFAT)途径对T2DM小鼠骨吸收代谢的影响。方法:采用6周高脂膳食和一次性注射链脲佐菌素(STZ)进行T2DM造模,成功后随机分为T2DM对照组(TC)、T2DM游泳组(TS)和T2DM下坡跑组(TD),另选C57小鼠为正常对照组(ZC)。T2DM小鼠继续高脂膳食,ZC小鼠饲以普通饲料。TS和TD小鼠分别进行8周游泳和下坡跑训练。末次训练24 h后处死小鼠并取材,应用Micro-CT、细胞原代培养、ELISA、RT-PCR及West-blotting等技术方法对骨组织形态计量学指标、OC数量、离子浓度、细胞因子m RNA和蛋白表达等进行检测。结果:TC组股骨中TRAF6、CN、Src-3、PLC、NFATc1、TRAP m RNA及胫骨中Src1和NFATc1蛋白表达上调(P<0.05),血清IP3和Ca2+浓度升高(P<0.05),BMM分化产生的OC总数量和≥10个核OC数量增多(P<0.01)。股骨远端松质骨和皮质骨骨组织形态计量学指标显著下降(P<0.05)。与TC比,TS组股骨中TRAF6、CN、PLC和TRAPm RNA及Src1蛋白表达下调,血清Ca2+浓度下降(P<0.05或P<0.01)。TD组股骨中TRAF6、CN、Src-3、PLC、NFATc1和TRAPm RNA及胫骨中Src1和NFATc1蛋白表达下调,血清IP3和Ca2+浓度下降(P<0.05)。OC总数量和≥10个核OC数量显著减少(P<0.05),松质骨和皮质骨骨组织形态计量学指标显著改善(P<0.05)。与TS比,TD组股骨中TRAF6、Src-3、PLC和TRAP m RNA表达下调及血清IP3和Ca^(2+)浓度下降(P<0.05),OC总数量(P<0.05)下降,松质骨BS/TV增加(P<0.05)。结论:T2DM小鼠骨吸收增强。下坡跑通过抑制T2DM小鼠骨中CN/NFAT途径,减少OC数量,降低骨吸收,改善骨组织形态结构,且其作用效果优于游泳。展开更多
目的:基于活化T细胞核因子2(NFAT2)/环氧化酶-2(COX-2)通路探讨雷公藤多苷片(TWPT)防治糖尿病肾病(DN)肾脏损伤的可能作用机制。方法:选取雄性清洁级SD大鼠42只,适应性喂养1周后随机分为正常组8只,造模组34只。正常组予以正常饲养,造模...目的:基于活化T细胞核因子2(NFAT2)/环氧化酶-2(COX-2)通路探讨雷公藤多苷片(TWPT)防治糖尿病肾病(DN)肾脏损伤的可能作用机制。方法:选取雄性清洁级SD大鼠42只,适应性喂养1周后随机分为正常组8只,造模组34只。正常组予以正常饲养,造模组采用高脂高糖饮食喂养1周后予腹腔注射链脲佐菌素(STZ)法建立DN大鼠模型,除去造模过程中死亡及失败,选取造模成功的24只随机分为模型组、缬沙坦(8.33 mg·kg^(-1)·d^(-1))组、TWPT(5 mg·kg^(-1)·d^(-1))组。正常组和模型组均予等体积生理盐水灌胃,6周后测量体质量,收集大鼠尿液,腹主动脉取血后处死取材,生化检测血清中的尿素氮(BUN)、肌酐(SCr)、丙氨酸氨基转移酶(ALT)、血脂血糖及尿液中的24 h尿蛋白总量(24 h UTP),苏木素-伊红(HE)及马松(Masson)染色观察肾脏病理,酶联免疫吸附测定法(ELISA)检测血清中的NFAT2、COX-2表达水平,蛋白免疫印迹法(Western blot)检测肾组织中NFAT2、COX-2蛋白表达,实时荧光定量聚合酶链式反应(Real-time PCR)检测肾组织中NFAT2、COX-2 mRNA表达。结果:与正常组比较,模型组大鼠24 h UTP、BUN、SCr、CHO、TG、FBG及血清NFAT2、COX-2表达显著升高(P<0.01),肾组织中的NFAT2、COX-2蛋白及mRNA表达显著升高(P<0.01),肾脏病理示肾小球体积增大,系膜细胞轻度增生,系膜基质增宽;与模型组比较,TWPT组大鼠24 h UTP、BUN、SCr、CHO、TG、FBG均明显降低(P<0.05,P<0.01);肾脏病理示肾小球形态基本正常,血清中NFAT2、COX-2表达显著降低(P<0.01),肾组织中的NFAT2、COX-2 mRNA和蛋白表达显著下降(P<0.01)。结论:TWPT可减轻DN大鼠24 h UTP、保护肾功能、改善肾脏病理,其作用机制可能与下调血清及肾组织NFAT2/COX-2表达相关。展开更多
基金Supported by the Health Research Foundation of Hunan Provincial Health Commission,No.W20243192Natural Science Foundation of Changsha,No.kq2403086+1 种基金National Natural Science Foundation of China,No.81571784Hunan Provincial Health Commission Hunan Provincial High-level Health Talent Major Scientific Research Project,No.R2023022.
文摘BACKGROUND Hepatic stellate cell(HSC)activation is key to liver fibrosis.Targeting DNA methylation shows promise.Zebularine,a methylation inhibitor,may suppress HSC activation via the calcineurin(CaN)/NFAT3 pathway.Magnetic resonance imaging(MRI)is a noninvasive tool for evaluating liver fibrosis evaluation tool,but multiparametric MRI for zebularine’s effects in liver fibrosis mouse models has not been studied.AIM To clarify the anti-fibrosis mechanism and MRI-evaluated efficacy of zebularine.METHODS In vitro,transforming growth factor(TGF)-β1-stimulated human HSCs(LX-2)were treated with zebularine.α-smooth muscle actin,fibrotic and anti-fibrotic gene levels,and regulator of calcineurin1(RCAN1)regulation were measured.In vivo,carbon tetrachloride(CCl_(4))-induced liver fibrosis in mice was treated with zebularine,and fibrosis was evaluated using various biochemical,histopathological,and MRI methods.RESULTS Zebularine upregulated RCAN1.4 protein(P<0.01)and inhibited the CaN/NFAT3 pathway(P<0.05).In HSCs,TGF-β1 reduced anti-fibrotic gene massage RNA(mRNA)and increased fibrotic mRNA(P<0.05),whereas zebularine had the opposite effects(P<0.01,P<0.05).CCl4-treated mice exhibited increases in various fibrosis-related indices,all of which were reversed by zebularine treatment(P<0.05).CONCLUSION Zebularine may reduce LX-2 activation and extracellular matrix deposition via RCAN1.4 and CaN/NFAT3 path-ways.Multiparametric MRI can assess its efficacy,suggesting zebularine’s potential as a liver fibrosis treatment.
文摘目的:探究游泳和下坡跑通过钙调磷酸酶(CN)/活化T细胞核因子(NFAT)途径对T2DM小鼠骨吸收代谢的影响。方法:采用6周高脂膳食和一次性注射链脲佐菌素(STZ)进行T2DM造模,成功后随机分为T2DM对照组(TC)、T2DM游泳组(TS)和T2DM下坡跑组(TD),另选C57小鼠为正常对照组(ZC)。T2DM小鼠继续高脂膳食,ZC小鼠饲以普通饲料。TS和TD小鼠分别进行8周游泳和下坡跑训练。末次训练24 h后处死小鼠并取材,应用Micro-CT、细胞原代培养、ELISA、RT-PCR及West-blotting等技术方法对骨组织形态计量学指标、OC数量、离子浓度、细胞因子m RNA和蛋白表达等进行检测。结果:TC组股骨中TRAF6、CN、Src-3、PLC、NFATc1、TRAP m RNA及胫骨中Src1和NFATc1蛋白表达上调(P<0.05),血清IP3和Ca2+浓度升高(P<0.05),BMM分化产生的OC总数量和≥10个核OC数量增多(P<0.01)。股骨远端松质骨和皮质骨骨组织形态计量学指标显著下降(P<0.05)。与TC比,TS组股骨中TRAF6、CN、PLC和TRAPm RNA及Src1蛋白表达下调,血清Ca2+浓度下降(P<0.05或P<0.01)。TD组股骨中TRAF6、CN、Src-3、PLC、NFATc1和TRAPm RNA及胫骨中Src1和NFATc1蛋白表达下调,血清IP3和Ca2+浓度下降(P<0.05)。OC总数量和≥10个核OC数量显著减少(P<0.05),松质骨和皮质骨骨组织形态计量学指标显著改善(P<0.05)。与TS比,TD组股骨中TRAF6、Src-3、PLC和TRAP m RNA表达下调及血清IP3和Ca^(2+)浓度下降(P<0.05),OC总数量(P<0.05)下降,松质骨BS/TV增加(P<0.05)。结论:T2DM小鼠骨吸收增强。下坡跑通过抑制T2DM小鼠骨中CN/NFAT途径,减少OC数量,降低骨吸收,改善骨组织形态结构,且其作用效果优于游泳。
文摘目的:基于活化T细胞核因子2(NFAT2)/环氧化酶-2(COX-2)通路探讨雷公藤多苷片(TWPT)防治糖尿病肾病(DN)肾脏损伤的可能作用机制。方法:选取雄性清洁级SD大鼠42只,适应性喂养1周后随机分为正常组8只,造模组34只。正常组予以正常饲养,造模组采用高脂高糖饮食喂养1周后予腹腔注射链脲佐菌素(STZ)法建立DN大鼠模型,除去造模过程中死亡及失败,选取造模成功的24只随机分为模型组、缬沙坦(8.33 mg·kg^(-1)·d^(-1))组、TWPT(5 mg·kg^(-1)·d^(-1))组。正常组和模型组均予等体积生理盐水灌胃,6周后测量体质量,收集大鼠尿液,腹主动脉取血后处死取材,生化检测血清中的尿素氮(BUN)、肌酐(SCr)、丙氨酸氨基转移酶(ALT)、血脂血糖及尿液中的24 h尿蛋白总量(24 h UTP),苏木素-伊红(HE)及马松(Masson)染色观察肾脏病理,酶联免疫吸附测定法(ELISA)检测血清中的NFAT2、COX-2表达水平,蛋白免疫印迹法(Western blot)检测肾组织中NFAT2、COX-2蛋白表达,实时荧光定量聚合酶链式反应(Real-time PCR)检测肾组织中NFAT2、COX-2 mRNA表达。结果:与正常组比较,模型组大鼠24 h UTP、BUN、SCr、CHO、TG、FBG及血清NFAT2、COX-2表达显著升高(P<0.01),肾组织中的NFAT2、COX-2蛋白及mRNA表达显著升高(P<0.01),肾脏病理示肾小球体积增大,系膜细胞轻度增生,系膜基质增宽;与模型组比较,TWPT组大鼠24 h UTP、BUN、SCr、CHO、TG、FBG均明显降低(P<0.05,P<0.01);肾脏病理示肾小球形态基本正常,血清中NFAT2、COX-2表达显著降低(P<0.01),肾组织中的NFAT2、COX-2 mRNA和蛋白表达显著下降(P<0.01)。结论:TWPT可减轻DN大鼠24 h UTP、保护肾功能、改善肾脏病理,其作用机制可能与下调血清及肾组织NFAT2/COX-2表达相关。
