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
OBJECTIVE To investigate icariside(ICS)Ⅱ protects against PC12 cel damage induced by oxygen-glucose deprivation and reoxygenation and explore its mechanism.METHODS The oxidative stress injury model was induced by oxy...OBJECTIVE To investigate icariside(ICS)Ⅱ protects against PC12 cel damage induced by oxygen-glucose deprivation and reoxygenation and explore its mechanism.METHODS The oxidative stress injury model was induced by oxygen-glucose deprivation/reoxygenation(OGD/R) 2 h/24 h in PC12 cells.N-acetyl-lcysteine(NAC),a classical anti-oxidant,was used as positive control.Pharmacodynamic experimental study groups as follows:control,control+ICS Ⅱ50 μmol·L^(-1),OGD/R,OGD/R+ICSⅡ 12.5 μmol·L^(-1),OGD/R + ICS Ⅱ 25 μmol·L^(-1),OGD/R + ICS Ⅱ50 μmol·L^(-1),and OGD/R+NAC 100 μmol·L^(-1) groups.Cell viability and lactate dehydrogenase(LDH) leakage rate were measured by MTT assay and LDH ELISA kit,respectively.Moreover,reactive oxygen species(ROS) ELISA kit was used for detection of intracellular ROS generation,Mito-SOX fluorescence staining was used for detecting production of ROS in mitochondria and mitochondrial membrane potential(MMP)was detected by rhodamine 123 dye.In addition,PC12 cells apoptosis was detected by one-step TUNEL assay.Furthermore,the expressions of nuclear factor erythroid 2-related factors(Nrf2),Keap1,HO^(-1),NQO^(-1),silent information regulator 3(SIRT3),IDH2,Bax,Bcl-2 and caspase 3 were detected by Western blotting analysis.RESULTS The results of MTT and LDH assay showed that OGD/R reduced the cell viability and improved LDH release compared with the control or ICSⅡ 50 μmol·L^(-1) alone(P<0.01).Meanwhile,OGD/R not only increased intracellular and mitochondrial ROS generation,but also elevated the fluorescence intensity of TUNEL staining,at the same time,the MMP was declined when challenged by OGD/R.Furthermore,the Western blotting results showed that OGD/R induced the increase in the expression of cytoplasm-Nrf2,Keap1,Bax and cleaved-caspase 3 level,while the decrease in the expression of nucleus-Nrf2,HO^(-1),NQO^(-1),SIRT3,IDH2 and Bcl-2(P<0.05).However,ICS Ⅱ significantly increased the viability of PC12 cells and reduced LDH leakage(P<0.01).Notably,ICS Ⅱ also suppressed ROS generation both in the intracellular and mitochondria,as well as restored MMP.It was also worthy to note that ICS Ⅱ decreased the expressions of cytoplasmNrf2,Keap1,Bax and the level of cleaved-caspase3,whereas,it increased the expressions of nucleus-Nrf2,HO^(-1),NQO^(-1),SIRT3,IDH2 and Bcl-2(P<0.05).CONCLUSION ICSⅡ reduced OGD/Rinduced oxidative damage in PC12 cells under the laboratory conditions,and its underlying mechanism may be related to the regulation of Nrf2/SIRT3 signaling pathway.展开更多
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。
基金National Natural Science Foundation of China(81560666)Program for Excellent Young Talents of Zunyi Medical Uiverstity(15zy-002)+1 种基金Science and Technology Innovation Talent Team of Guizhou Province(20154023)the ″Hundred″Level of High-level Innovative Talents in Guizhou Province(QKHRCPT 20165684)；and Program forChangjiang Scholars and Innovative ResearchTeam in University of China(IRT一17R113).
文摘OBJECTIVE To investigate icariside(ICS)Ⅱ protects against PC12 cel damage induced by oxygen-glucose deprivation and reoxygenation and explore its mechanism.METHODS The oxidative stress injury model was induced by oxygen-glucose deprivation/reoxygenation(OGD/R) 2 h/24 h in PC12 cells.N-acetyl-lcysteine(NAC),a classical anti-oxidant,was used as positive control.Pharmacodynamic experimental study groups as follows:control,control+ICS Ⅱ50 μmol·L^(-1),OGD/R,OGD/R+ICSⅡ 12.5 μmol·L^(-1),OGD/R + ICS Ⅱ 25 μmol·L^(-1),OGD/R + ICS Ⅱ50 μmol·L^(-1),and OGD/R+NAC 100 μmol·L^(-1) groups.Cell viability and lactate dehydrogenase(LDH) leakage rate were measured by MTT assay and LDH ELISA kit,respectively.Moreover,reactive oxygen species(ROS) ELISA kit was used for detection of intracellular ROS generation,Mito-SOX fluorescence staining was used for detecting production of ROS in mitochondria and mitochondrial membrane potential(MMP)was detected by rhodamine 123 dye.In addition,PC12 cells apoptosis was detected by one-step TUNEL assay.Furthermore,the expressions of nuclear factor erythroid 2-related factors(Nrf2),Keap1,HO^(-1),NQO^(-1),silent information regulator 3(SIRT3),IDH2,Bax,Bcl-2 and caspase 3 were detected by Western blotting analysis.RESULTS The results of MTT and LDH assay showed that OGD/R reduced the cell viability and improved LDH release compared with the control or ICSⅡ 50 μmol·L^(-1) alone(P<0.01).Meanwhile,OGD/R not only increased intracellular and mitochondrial ROS generation,but also elevated the fluorescence intensity of TUNEL staining,at the same time,the MMP was declined when challenged by OGD/R.Furthermore,the Western blotting results showed that OGD/R induced the increase in the expression of cytoplasm-Nrf2,Keap1,Bax and cleaved-caspase 3 level,while the decrease in the expression of nucleus-Nrf2,HO^(-1),NQO^(-1),SIRT3,IDH2 and Bcl-2(P<0.05).However,ICS Ⅱ significantly increased the viability of PC12 cells and reduced LDH leakage(P<0.01).Notably,ICS Ⅱ also suppressed ROS generation both in the intracellular and mitochondria,as well as restored MMP.It was also worthy to note that ICS Ⅱ decreased the expressions of cytoplasmNrf2,Keap1,Bax and the level of cleaved-caspase3,whereas,it increased the expressions of nucleus-Nrf2,HO^(-1),NQO^(-1),SIRT3,IDH2 and Bcl-2(P<0.05).CONCLUSION ICSⅡ reduced OGD/Rinduced oxidative damage in PC12 cells under the laboratory conditions,and its underlying mechanism may be related to the regulation of Nrf2/SIRT3 signaling pathway.
