目的探讨E3泛素连接酶32(Trim32)调控血管平滑肌细胞(VSMC)增殖与迁移的作用机制。方法选择C57雄性野生型小鼠40只,其中10只小鼠从胸主动脉获得VSMC后,分为空白组、对照组、转染组、模型组、对照1组、转染1组和激动剂组(β-catenin激动...目的探讨E3泛素连接酶32(Trim32)调控血管平滑肌细胞(VSMC)增殖与迁移的作用机制。方法选择C57雄性野生型小鼠40只,其中10只小鼠从胸主动脉获得VSMC后,分为空白组、对照组、转染组、模型组、对照1组、转染1组和激动剂组(β-catenin激动剂同时处理),后4组用重组人血小板衍生生长因子30 mg/L干预24 h,再按分组分别转染阴性干扰siControl和siTrim32,用PCR和Western blot检测各组Trim32、β-catenin mRNA和蛋白表达。观察各组细胞增殖和迁移情况。另30只小鼠左颈动脉损伤处理,取右、左颈动脉分为假手术组、假手术对照组、假手术转染组、损伤组、损伤对照组和损伤转染组,每组10只,按分组处理后,检测Trim32表达和血管内膜增生情况。结果与空白组比较,模型组Trim32 mRNA和蛋白表达升高(1.89±0.30 vs 0.99±0.06,1.82±0.30 vs 1.02±0.05,P<0.01)。与对照组比较,转染组细胞增殖、阳性细胞、β-catenin蛋白表达降低,对照1组各项明显升高(P<0.05,P<0.01),与转染1组比较,对照1组和激动剂组阳性细胞升高(P<0.01)。与假手术组比较,损伤组Trim32 mRNA和蛋白明显升高(P<0.01)。与损伤对照组比较,假手术对照组和损伤转染组内膜/中膜面积比值降低(P<0.01),损伤转染组内膜/中膜面积比值较假手术转染组升高(2.11±0.64 vs 0.37±0.04,P<0.05)。结论Trim32可能通过β-catenin途径促进VSMC增殖及迁移。展开更多
OBJECTIVE:To assess the protective role of benazepril,an angiotensin-converting enzyme inhibitor,in renal damage caused by prenatal inflammation.METHODS:Saline or lipopolysaccharide were administered intraperitoneally...OBJECTIVE:To assess the protective role of benazepril,an angiotensin-converting enzyme inhibitor,in renal damage caused by prenatal inflammation.METHODS:Saline or lipopolysaccharide were administered intraperitoneally to pregnant Sprague-Dawley rats on gestation days 8,10,and 12.After birth,offspring received either tap water or benazepril in water between 7 and 68 weeks.Blood pressure,blood urea nitrogen,creatinine,and 24-h urine volume were measured as indices of renal function.Hematoxylin,eosin,periodic acid-Schiff,and Sirius Red staining were used to evaluate renal damage.RESULTS:Postnatal benazepril treatment ameliorated hypertension and restored normal 24-h urine volume and blood urea nitrogen and serum creatinine levels.Benazepril treatment also reduced glycoprotein accumulation and fibrosis in the glomerulus and in tubular epithelial cells and inhibited nuclear factor-kappa B activation.CONCLUSION:Together with our previous findings that postnatal inhibition of nuclear factor-kappa B activation blocks intra-renal renin-angiotensin system activation,our current data demonstrate that intra-renal activation of the renin-angiotensin system interacts with nuclear factor-kappa B activation to cause renal damage in adulthood following prenatal inflammation.展开更多
文摘目的探讨E3泛素连接酶32(Trim32)调控血管平滑肌细胞(VSMC)增殖与迁移的作用机制。方法选择C57雄性野生型小鼠40只,其中10只小鼠从胸主动脉获得VSMC后,分为空白组、对照组、转染组、模型组、对照1组、转染1组和激动剂组(β-catenin激动剂同时处理),后4组用重组人血小板衍生生长因子30 mg/L干预24 h,再按分组分别转染阴性干扰siControl和siTrim32,用PCR和Western blot检测各组Trim32、β-catenin mRNA和蛋白表达。观察各组细胞增殖和迁移情况。另30只小鼠左颈动脉损伤处理,取右、左颈动脉分为假手术组、假手术对照组、假手术转染组、损伤组、损伤对照组和损伤转染组,每组10只,按分组处理后,检测Trim32表达和血管内膜增生情况。结果与空白组比较,模型组Trim32 mRNA和蛋白表达升高(1.89±0.30 vs 0.99±0.06,1.82±0.30 vs 1.02±0.05,P<0.01)。与对照组比较,转染组细胞增殖、阳性细胞、β-catenin蛋白表达降低,对照1组各项明显升高(P<0.05,P<0.01),与转染1组比较,对照1组和激动剂组阳性细胞升高(P<0.01)。与假手术组比较,损伤组Trim32 mRNA和蛋白明显升高(P<0.01)。与损伤对照组比较,假手术对照组和损伤转染组内膜/中膜面积比值降低(P<0.01),损伤转染组内膜/中膜面积比值较假手术转染组升高(2.11±0.64 vs 0.37±0.04,P<0.05)。结论Trim32可能通过β-catenin途径促进VSMC增殖及迁移。
基金Natural Science Foundation-funded Project:Role of HMGB-1-TLR4/RAGE Pathway Over-Expression in Increased Offspring's Pyelonephritis Sensitivity Induced by Pregnant Inflammatory Stimulation(No.81703522)the Role and Mechanisms of Decidual NK Cell Abnormality in Prenatal Inflammation-Induced Offspring's Immune System Dysfunction and Hypertension(No.81520108029)+1 种基金Role of AMPK/NK-ΚB Signal Pathway Imbalance in Offspring's Hypertension Induced by Pregnant Inflammatory Stimulation(No.81703521)Roles and Mechanisms of Mitochondrial Dyshomeostasis in the Hypersensitivity of Cardiac Damage Responding to Cardiac Risk Factors in Offspring of Prenatal Inflammatory Stimulation(No.81773742)。
文摘OBJECTIVE:To assess the protective role of benazepril,an angiotensin-converting enzyme inhibitor,in renal damage caused by prenatal inflammation.METHODS:Saline or lipopolysaccharide were administered intraperitoneally to pregnant Sprague-Dawley rats on gestation days 8,10,and 12.After birth,offspring received either tap water or benazepril in water between 7 and 68 weeks.Blood pressure,blood urea nitrogen,creatinine,and 24-h urine volume were measured as indices of renal function.Hematoxylin,eosin,periodic acid-Schiff,and Sirius Red staining were used to evaluate renal damage.RESULTS:Postnatal benazepril treatment ameliorated hypertension and restored normal 24-h urine volume and blood urea nitrogen and serum creatinine levels.Benazepril treatment also reduced glycoprotein accumulation and fibrosis in the glomerulus and in tubular epithelial cells and inhibited nuclear factor-kappa B activation.CONCLUSION:Together with our previous findings that postnatal inhibition of nuclear factor-kappa B activation blocks intra-renal renin-angiotensin system activation,our current data demonstrate that intra-renal activation of the renin-angiotensin system interacts with nuclear factor-kappa B activation to cause renal damage in adulthood following prenatal inflammation.
