Objective:To investigate the effect of Qihuang Bushen Xiezhuo Formula on the expression of the nuclear factor E2 related factor 2(Nrf2)/antioxidant response element(ARE)signaling pathway of human glomerular mesangial ...Objective:To investigate the effect of Qihuang Bushen Xiezhuo Formula on the expression of the nuclear factor E2 related factor 2(Nrf2)/antioxidant response element(ARE)signaling pathway of human glomerular mesangial cells(HMC)in high glucose medium and its protective effect on oxidative stress.Methods:The HMC were cultured in vitro to prepare normal rat serum.The rats were intragastrically administered with irbesartan and Qihuang Bushen Xiezhuo Formula.The serum containing the drugs was prepared after the blood concentration was reached.The rats were divided into the control group,the high glucose group,the irbesartan group and the Qihuang Bushen Xiezhuo Formula group.The HMC of the control group was cultured with normal rat serum(10%serum concentration)medium,while that of the high glucose group was cultured with high glucose medium of rat serum(10%serum concentration).The irbesartan group and the Qihuang Bushen Xiezhuo Formula group were respectively treated with 10%irbesartan and 10%Qihuang Bushen Xiezhuo Formula in serum high glucose medium.After 48 h of culture,the relevant indicators were collected and detected.The mRNA expression levels of Nrf2,gamma-glutamylcysteine synthetase(γ-GCS)and superoxide dismutase(SOD)in each group were detected by real-time PCR.The expressions ofγ-GCS and SOD were detected by immunohistochemistry.The protein expressions ofγ-GCS and SOD in HMC of each group were observed by Western Blot.Results:The expressions of Nrf2,γ-GCS,SOD mRNA and protein in experimental cells of each group were low in the control group,while those in the high glucose group were decreased as compared with the control group(P<0.05).After interference of irbesartan and Qihuang Bushen Xiezhuo Formula the expressions were increased,and the increase in Qihuang Bushen Xiezhuo Formula group was more significant(P<0.05).Conclusion:Qihuang Bushen Xiezhuo Formula can improve HMC oxidative stress injury in high glucose culture,and its mechanism may be achieved by activating Nrf2 and its related downstream proteinsγ-GCS and SOD.展开更多
目的:探讨小豆蔻明抑制C3H10T1/2细胞成脂分化的作用机制。方法:观察C3H10T1/2细胞成脂过程形态学变化,采用MTS法、油红O染色法、GAP-PAP酶法检测小豆蔻明对C3H10T1/2细胞增殖、成脂分化以及胞内甘油三酯含量的影响,采用RT-PCR与Western...目的:探讨小豆蔻明抑制C3H10T1/2细胞成脂分化的作用机制。方法:观察C3H10T1/2细胞成脂过程形态学变化,采用MTS法、油红O染色法、GAP-PAP酶法检测小豆蔻明对C3H10T1/2细胞增殖、成脂分化以及胞内甘油三酯含量的影响,采用RT-PCR与Western Blot技术检测小豆蔻明对C3H10T1/2细胞PPARγ、C/EBPα和FABP4 m RNA以及蛋白表达的影响。结果:60、80、100μmol/L小豆蔻明能够显著抑制C3H10T1/2细胞存活率(P<0.01);5、10、30μmol/L小豆蔻明能够抑制C3H10T1/2细胞脂滴的形成,从而抑制成脂分化;5、10、30μmol/L小豆蔻明能够显著降低细胞中甘油三酯的含量(P<0.05,P<0.01);10、30μmol/L小豆蔻明能够显著下调PPARγ、C/EBPα和FABP4 m RNA的表达(P<0.05,P<0.01);5、10、30μmol/L小豆蔻明能够显著降低成脂关键转录因子PPARγ、C/EBPα和成脂分化标志因子FABP4蛋白的表达(P<0.05,P<0.01)。结论:小豆蔻明通过下调PPARγ、C/EBPα、FABP4 m RNA以及蛋白表达抑制细胞成脂分化并降低胞内甘油三酯含量。展开更多
基金General Project of Natural Science Foundation of Heilongjiang Province(No.H2016066)Young Chinese Medicine Science and Technology Innovation Project of Heilongjiang Chinese Medicine Association(No.ZHY19-023)。
文摘Objective:To investigate the effect of Qihuang Bushen Xiezhuo Formula on the expression of the nuclear factor E2 related factor 2(Nrf2)/antioxidant response element(ARE)signaling pathway of human glomerular mesangial cells(HMC)in high glucose medium and its protective effect on oxidative stress.Methods:The HMC were cultured in vitro to prepare normal rat serum.The rats were intragastrically administered with irbesartan and Qihuang Bushen Xiezhuo Formula.The serum containing the drugs was prepared after the blood concentration was reached.The rats were divided into the control group,the high glucose group,the irbesartan group and the Qihuang Bushen Xiezhuo Formula group.The HMC of the control group was cultured with normal rat serum(10%serum concentration)medium,while that of the high glucose group was cultured with high glucose medium of rat serum(10%serum concentration).The irbesartan group and the Qihuang Bushen Xiezhuo Formula group were respectively treated with 10%irbesartan and 10%Qihuang Bushen Xiezhuo Formula in serum high glucose medium.After 48 h of culture,the relevant indicators were collected and detected.The mRNA expression levels of Nrf2,gamma-glutamylcysteine synthetase(γ-GCS)and superoxide dismutase(SOD)in each group were detected by real-time PCR.The expressions ofγ-GCS and SOD were detected by immunohistochemistry.The protein expressions ofγ-GCS and SOD in HMC of each group were observed by Western Blot.Results:The expressions of Nrf2,γ-GCS,SOD mRNA and protein in experimental cells of each group were low in the control group,while those in the high glucose group were decreased as compared with the control group(P<0.05).After interference of irbesartan and Qihuang Bushen Xiezhuo Formula the expressions were increased,and the increase in Qihuang Bushen Xiezhuo Formula group was more significant(P<0.05).Conclusion:Qihuang Bushen Xiezhuo Formula can improve HMC oxidative stress injury in high glucose culture,and its mechanism may be achieved by activating Nrf2 and its related downstream proteinsγ-GCS and SOD.
文摘目的:探讨小豆蔻明抑制C3H10T1/2细胞成脂分化的作用机制。方法:观察C3H10T1/2细胞成脂过程形态学变化,采用MTS法、油红O染色法、GAP-PAP酶法检测小豆蔻明对C3H10T1/2细胞增殖、成脂分化以及胞内甘油三酯含量的影响,采用RT-PCR与Western Blot技术检测小豆蔻明对C3H10T1/2细胞PPARγ、C/EBPα和FABP4 m RNA以及蛋白表达的影响。结果:60、80、100μmol/L小豆蔻明能够显著抑制C3H10T1/2细胞存活率(P<0.01);5、10、30μmol/L小豆蔻明能够抑制C3H10T1/2细胞脂滴的形成,从而抑制成脂分化;5、10、30μmol/L小豆蔻明能够显著降低细胞中甘油三酯的含量(P<0.05,P<0.01);10、30μmol/L小豆蔻明能够显著下调PPARγ、C/EBPα和FABP4 m RNA的表达(P<0.05,P<0.01);5、10、30μmol/L小豆蔻明能够显著降低成脂关键转录因子PPARγ、C/EBPα和成脂分化标志因子FABP4蛋白的表达(P<0.05,P<0.01)。结论:小豆蔻明通过下调PPARγ、C/EBPα、FABP4 m RNA以及蛋白表达抑制细胞成脂分化并降低胞内甘油三酯含量。
