Objective: To demonstrate whether the expression of silent mating type information regulation 2 homolog 1 (SIRT1) affects the level of TGF-β1 and Smad3 in HEK293 cells through regulating mTOR. Methods: First, recombi...Objective: To demonstrate whether the expression of silent mating type information regulation 2 homolog 1 (SIRT1) affects the level of TGF-β1 and Smad3 in HEK293 cells through regulating mTOR. Methods: First, recombinant plasmids DNA (rSIRT1) and siRNA targeting SIRT1 were constructed which were transfected into Human Embryonic Kidney 293 cell (HEK293) cells, respectively. Then, the generation of intracellular ROS in cells was examined by flow cytometry using the oxidation-sensitive probe. Last, the expressions of TGF-β1, smad3, P53, mTOR, p-mTOR, LC3-I and LC3-II in cells were detected to observe the effect of SIRT1 on TGF-β1 Pathway by western blot analysis. Results: We demonstrated that overexpressing of SIRT1 may decrease TGF-β1 and Smad3 expression in HEK293 cells through regulating mTOR. In addition, the result is the opposite when SIRT1 was silent in HEK293 cells. Conclusions: SIRT1 is closely related to TGF-β1/Smad3 pathway that correlates with the regulation of mTOR and ROS generation and causes diabetic nephropathy. The available evidence implies that SIRT1 has great potential as a clinical target for the prevention and treatment of renal fibrosis in the development of DN.展开更多
AIM: To detect the expression of miR-211 in age-related cataract tissue, explore the effects of miR-211 on lens epithelial cell proliferation and apoptosis, and identify its target gene.METHODS: This study used real...AIM: To detect the expression of miR-211 in age-related cataract tissue, explore the effects of miR-211 on lens epithelial cell proliferation and apoptosis, and identify its target gene.METHODS: This study used real-time quantitative polymerase chain reaction(RT-q PCR) to measure the expression of miR-211 and its predicted target gene [silent matingtype information regulation 2 homolog 1(SIRT1)] in 46 anterior lens capsules collected from age-related cataract patients. Human lens epithelial cell line(SRA01/04) cells were transfected with either miR-211 mimics, mimic controls, miR-211 inhibitors or inhibitor controls, 72 h after transfection, miR NA and protein expression of SIRT1 were measured using RT-qP CR and Western blotting; then cells were exposed to 200 μmol/L H2O2 for 1h, whereupon cell viability was measured by MTS assay, caspase-3 assay was performed. Dual luciferase reporter assay was performed to verify the relationship between miR-211 of SIRT1.RESULTS: Compared to the control group, expression of miR-211 was significantly increased(P〈0.001), the miR NA and protein expression of SIRT1 were significantly decreased(P〈0.001) in the anterior lens capsules of patients with age-related cataracts. Relative to the control group, SIRT1 miR NA and protein levels in the miR-211 mimic group were significantly reduced, cell proliferation activity significantly decreased, and caspase-3 activity was significantly increased(P〈0.001). In the miR-211 inhibitor group, SIRT1 miRNA and protein expression were significantly increased, cell proliferation activity significantly increased, and caspase-3 activity was significantly decreased(P〈0.001). A dual luciferase reporter assay confirmed that SIRT1 is a direct target of miR-211.CONCLUSION: miR-211 is highly expressed in the anterior lens capsules of patients with age-related cataracts. By negatively regulating the expression of SIRT1, miR-211 promotes lens epithelial cell apoptosis and inhibits lens epithelial cell proliferation.展开更多
沉默信息调节因子2同源体1(silent mating type information regulation 2 homolog 1,SIRT1)基因作为一种编码组蛋白去乙酰化酶的基因,在能量代谢、延缓衰老、抗肿瘤和对抗心脏疾病方面得到关注,更重要的是其对多种神经退行性疾病的作...沉默信息调节因子2同源体1(silent mating type information regulation 2 homolog 1,SIRT1)基因作为一种编码组蛋白去乙酰化酶的基因,在能量代谢、延缓衰老、抗肿瘤和对抗心脏疾病方面得到关注,更重要的是其对多种神经退行性疾病的作用引起了研究人员的兴趣。其激动剂白藜芦醇,是一种天然多酚类物质,也具有神经保护作用,作用途径广泛,如抗氧化,抗缺血等。SRT1720作为新合成的SIRT1激动剂,具有特异性强的优势,但其他方面作用尚待进一步研究。展开更多
目的探讨固本降消汤联合达格列净经腺苷酸活化蛋白激酶(AMPK)信号通路、沉默信息调节因子调治2型糖尿病(T2DM)伴肥胖的效果及机制。方法选取2021年1—12月收治的T2DM伴肥胖120例,根据治疗方案不同将其分为观察组和对照组2组各60例。观...目的探讨固本降消汤联合达格列净经腺苷酸活化蛋白激酶(AMPK)信号通路、沉默信息调节因子调治2型糖尿病(T2DM)伴肥胖的效果及机制。方法选取2021年1—12月收治的T2DM伴肥胖120例,根据治疗方案不同将其分为观察组和对照组2组各60例。观察组采用固本降消汤联合达格列净治疗,对照组采用达格列净治疗,连续治疗3个月。比较2组治疗3个月后临床效果,治疗前及治疗1、3个月后血糖指标[空腹血糖(FPG)、糖化血红蛋白(HbA1c)、餐后2 h血糖(2 h PG)]、氧化应激指标[丙二醛(MDA)、超氧化物歧化酶(ROS)、谷胱甘肽过氧化物酶(GSH-Px)]、腺苷酸活化蛋白激酶α1/Toll样受体4(AMPKα1/TLR4)、转化生长因子-β1(TGF-β1)、核因子-κB(NF-κB)、能量平衡相关蛋白(Adropin)、鸢尾素(Irisin)、趋化素(Chemerin)及沉默信息调节因子1(SIRT1)、沉默信息调节因子3(SIRT3),以及治疗期间不良反应发生情况。结果治疗3个月后,总有效率观察组为96.67%高于对照组83.33%(P<0.05)。治疗1和3个月后,FPG、HbA1c、2 h PG、MDA、ROS、TGF-β1、NF-κB及Chemerin 2组均较治疗前降低,且观察组低于对照组;GSH-Px、AMPKα1/TLR4、Adropin、Irisin及SIRT1、SIRT32组均较治疗前升高,且观察组高于对照组(P<0.05,P<0.01)。治疗期间,不良反应总发生率观察组为13.33%,对照组为10.00%,2组比较差异无统计学意义(P>0.05)。结论固本降消汤联合达格列净调治T2DM伴肥胖可提高临床效果,改善血糖水平,减轻氧化应激损伤,并可经AMPK信号通路抑制肾损伤,调节SIRT1和SIRT3,延缓病情进展。展开更多
文摘Objective: To demonstrate whether the expression of silent mating type information regulation 2 homolog 1 (SIRT1) affects the level of TGF-β1 and Smad3 in HEK293 cells through regulating mTOR. Methods: First, recombinant plasmids DNA (rSIRT1) and siRNA targeting SIRT1 were constructed which were transfected into Human Embryonic Kidney 293 cell (HEK293) cells, respectively. Then, the generation of intracellular ROS in cells was examined by flow cytometry using the oxidation-sensitive probe. Last, the expressions of TGF-β1, smad3, P53, mTOR, p-mTOR, LC3-I and LC3-II in cells were detected to observe the effect of SIRT1 on TGF-β1 Pathway by western blot analysis. Results: We demonstrated that overexpressing of SIRT1 may decrease TGF-β1 and Smad3 expression in HEK293 cells through regulating mTOR. In addition, the result is the opposite when SIRT1 was silent in HEK293 cells. Conclusions: SIRT1 is closely related to TGF-β1/Smad3 pathway that correlates with the regulation of mTOR and ROS generation and causes diabetic nephropathy. The available evidence implies that SIRT1 has great potential as a clinical target for the prevention and treatment of renal fibrosis in the development of DN.
基金Supported by the National Natural Science Foundation of China(No.81170836No.81570838)+1 种基金the Natural Science Foundation of Liaoning Province,China(No.2015020474)the Liaoning Provincial Hospital Program for Building Treatment Capacity in Key Clinical Departments(No.LNCCC-D15-2015)
文摘AIM: To detect the expression of miR-211 in age-related cataract tissue, explore the effects of miR-211 on lens epithelial cell proliferation and apoptosis, and identify its target gene.METHODS: This study used real-time quantitative polymerase chain reaction(RT-q PCR) to measure the expression of miR-211 and its predicted target gene [silent matingtype information regulation 2 homolog 1(SIRT1)] in 46 anterior lens capsules collected from age-related cataract patients. Human lens epithelial cell line(SRA01/04) cells were transfected with either miR-211 mimics, mimic controls, miR-211 inhibitors or inhibitor controls, 72 h after transfection, miR NA and protein expression of SIRT1 were measured using RT-qP CR and Western blotting; then cells were exposed to 200 μmol/L H2O2 for 1h, whereupon cell viability was measured by MTS assay, caspase-3 assay was performed. Dual luciferase reporter assay was performed to verify the relationship between miR-211 of SIRT1.RESULTS: Compared to the control group, expression of miR-211 was significantly increased(P〈0.001), the miR NA and protein expression of SIRT1 were significantly decreased(P〈0.001) in the anterior lens capsules of patients with age-related cataracts. Relative to the control group, SIRT1 miR NA and protein levels in the miR-211 mimic group were significantly reduced, cell proliferation activity significantly decreased, and caspase-3 activity was significantly increased(P〈0.001). In the miR-211 inhibitor group, SIRT1 miRNA and protein expression were significantly increased, cell proliferation activity significantly increased, and caspase-3 activity was significantly decreased(P〈0.001). A dual luciferase reporter assay confirmed that SIRT1 is a direct target of miR-211.CONCLUSION: miR-211 is highly expressed in the anterior lens capsules of patients with age-related cataracts. By negatively regulating the expression of SIRT1, miR-211 promotes lens epithelial cell apoptosis and inhibits lens epithelial cell proliferation.
文摘沉默信息调节因子2同源体1(silent mating type information regulation 2 homolog 1,SIRT1)基因作为一种编码组蛋白去乙酰化酶的基因,在能量代谢、延缓衰老、抗肿瘤和对抗心脏疾病方面得到关注,更重要的是其对多种神经退行性疾病的作用引起了研究人员的兴趣。其激动剂白藜芦醇,是一种天然多酚类物质,也具有神经保护作用,作用途径广泛,如抗氧化,抗缺血等。SRT1720作为新合成的SIRT1激动剂,具有特异性强的优势,但其他方面作用尚待进一步研究。
文摘目的探讨固本降消汤联合达格列净经腺苷酸活化蛋白激酶(AMPK)信号通路、沉默信息调节因子调治2型糖尿病(T2DM)伴肥胖的效果及机制。方法选取2021年1—12月收治的T2DM伴肥胖120例,根据治疗方案不同将其分为观察组和对照组2组各60例。观察组采用固本降消汤联合达格列净治疗,对照组采用达格列净治疗,连续治疗3个月。比较2组治疗3个月后临床效果,治疗前及治疗1、3个月后血糖指标[空腹血糖(FPG)、糖化血红蛋白(HbA1c)、餐后2 h血糖(2 h PG)]、氧化应激指标[丙二醛(MDA)、超氧化物歧化酶(ROS)、谷胱甘肽过氧化物酶(GSH-Px)]、腺苷酸活化蛋白激酶α1/Toll样受体4(AMPKα1/TLR4)、转化生长因子-β1(TGF-β1)、核因子-κB(NF-κB)、能量平衡相关蛋白(Adropin)、鸢尾素(Irisin)、趋化素(Chemerin)及沉默信息调节因子1(SIRT1)、沉默信息调节因子3(SIRT3),以及治疗期间不良反应发生情况。结果治疗3个月后,总有效率观察组为96.67%高于对照组83.33%(P<0.05)。治疗1和3个月后,FPG、HbA1c、2 h PG、MDA、ROS、TGF-β1、NF-κB及Chemerin 2组均较治疗前降低,且观察组低于对照组;GSH-Px、AMPKα1/TLR4、Adropin、Irisin及SIRT1、SIRT32组均较治疗前升高,且观察组高于对照组(P<0.05,P<0.01)。治疗期间,不良反应总发生率观察组为13.33%,对照组为10.00%,2组比较差异无统计学意义(P>0.05)。结论固本降消汤联合达格列净调治T2DM伴肥胖可提高临床效果,改善血糖水平,减轻氧化应激损伤,并可经AMPK信号通路抑制肾损伤,调节SIRT1和SIRT3,延缓病情进展。