山奈酚对高糖条件下人肾小球内皮细胞氧化应激及凋亡的影响
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摘要
目的观察山奈酚对高糖条件下人肾小球内皮细胞氧化应激及凋亡的影响。方法 2016年4月—2018年8月于陕西省人民医院中心实验室进行实验。培养人肾小球内皮细胞后分为5.5 mmol/L葡萄糖处理的对照组、30.0 mmol/L葡萄糖处理的高糖组、30.0 mmol/L葡萄糖+山奈酚50μmol/L处理的山奈酚组,测定活性氧(ROS)、凋亡率、氧化应激及凋亡基因的表达水平。结果高糖组的ROS含量、凋亡率及NOX2、NOX4、Nrf2、HO-1、Bax、VDAC1的mRNA表达水平均明显高于对照组(t/P=6.321/0.000、7.141/0.000、5.324/0.001、6.103/0.000、4.203/0.003、4.748/0.001、5.325/0.001、5.884/0.000),HK-II、Bcl-2的mRNA表达水平均明显低于对照组(t/P=7.125/0.000、6.718/0.000);山奈酚组的ROS含量、凋亡率及NOX2、NOX4、Bax、VDAC1的mRNA表达水平均明显低于高糖组(t/P=3.518/0.008、5.349/0.001、2.643/0.030、2.647/0.029、2.492/0.037、2.440/0.041),Nrf2、HO-1、HK-II、Bcl-2的mRNA表达水平均明显高于高糖组(t/P=3.308/0.011、2.833/0.022、4.828/0.001、3.923/0.000)。结论山奈酚对高糖条件下人肾小球内皮细胞氧化应激及凋亡具有抑制作用。
Objective To observe the effects of kaempferol on oxidative stress and apoptosis of human glomerular endothelial cells under high glucose. Methods The experiment was conducted in the Central Laboratory of Shaanxi People's Hospital from April 2016 to August 2018. The cultured human glomerular endothelial cells were divided into 5.5 mmol/L glucose-treated control group, 30.0 mmol/L glucose-treated high-sugar group, 30.0 mmol/L glucose + kaempferol 50 μmol/L-treated kaempferol group. The expression of reactive oxygen species(ROS), apoptotic rate, oxidative stress and apoptotic genes were measured. Results The ROS content, apoptotic rate and mRNA expression levels of NOX2, NOX4, Nrf2, HO-1, Bax and VDAC1 in the high glucose group were significantly higher than those in the control group(t/P=6.321/0.000,t/P=7.141/0.000,t/P=5.324/0.001,t/P=6.103/0.000,t/P=4.203/0.003,t/P=4.748/0.001,t/P=5.325/0.001,t/P=5.884/0.000), HK The mRNA expression levels of II and Bcl 2 were significantly lower than those of the control group(t/P=7.125/0.000,t/P=6.718/0.000); ROS content, apoptotic rate and NOX2, NOX4, Bax, VDAC1 in the kaempferol group The mRNA expression level was significantly lower than that of the high glucose group(t/P=3.518/0.008,t/P=5.349/0.001,t/P=2.643/0.030,t/P=2.647/0.029,t/P=2.492/0.037,t/P=2.440/0.041), mRNA expression levels of Nrf2, HO 1, HK II, Bcl 2 were significantly higher than those of high glucose group(t/P=3.308/0.011,t/P=2.833/0.022,t/P=4.828/0.001,t/P=3.923/0.000). Conclusion Kaempferol has an inhibitory effect on oxidative stress and apoptosis of human glomerular endothelial cells under high glucose conditions.
Objective To observe the effects of kaempferol on oxidative stress and apoptosis of human glomerular endothelial cells under high glucose. Methods The experiment was conducted in the Central Laboratory of Shaanxi People's Hospital from April 2016 to August 2018. The cultured human glomerular endothelial cells were divided into 5.5 mmol/L glucose-treated control group, 30.0 mmol/L glucose-treated high-sugar group, 30.0 mmol/L glucose + kaempferol 50 μmol/L-treated kaempferol group. The expression of reactive oxygen species(ROS), apoptotic rate, oxidative stress and apoptotic genes were measured. Results The ROS content, apoptotic rate and mRNA expression levels of NOX2, NOX4, Nrf2, HO-1, Bax and VDAC1 in the high glucose group were significantly higher than those in the control group(t/P=6.321/0.000,t/P=7.141/0.000,t/P=5.324/0.001,t/P=6.103/0.000,t/P=4.203/0.003,t/P=4.748/0.001,t/P=5.325/0.001,t/P=5.884/0.000), HK The mRNA expression levels of II and Bcl 2 were significantly lower than those of the control group(t/P=7.125/0.000,t/P=6.718/0.000); ROS content, apoptotic rate and NOX2, NOX4, Bax, VDAC1 in the kaempferol group The mRNA expression level was significantly lower than that of the high glucose group(t/P=3.518/0.008,t/P=5.349/0.001,t/P=2.643/0.030,t/P=2.647/0.029,t/P=2.492/0.037,t/P=2.440/0.041), mRNA expression levels of Nrf2, HO 1, HK II, Bcl 2 were significantly higher than those of high glucose group(t/P=3.308/0.011,t/P=2.833/0.022,t/P=4.828/0.001,t/P=3.923/0.000). Conclusion Kaempferol has an inhibitory effect on oxidative stress and apoptosis of human glomerular endothelial cells under high glucose conditions.
引文
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[13] Volpe CMO, Villar-Delfino PH, Dos Anjos PMF, et al. Cellular death, reactive oxygen species (ROS) and diabetic complications[J]. Cell Death Dis, 2018, 9(2):119. DOI: 10.1038/s41419-017-0135-z.
[14] 张微,熊军,王景平,叶红,李莉.缬沙坦联合前列地尔对慢性肾小球肾炎患者肾功能、免疫指标及其细胞因子的影响[J].疑难病杂志,2017,16(12):1236-1239.DOI:10.3969/j.issn.1671-6450.2017.12.013
[15] Sha J, Sui B, Su X, et al. Alteration of oxidative stress and inflammatory cytokines induces apoptosis in diabetic nephropathy[J]. Mol Med Rep, 2017, 16(5):7715-7723. DOI: 10.3892/mmr.2017.7522.
[16] Zhang J, Guo Y, Ge W,et al. High glucose induces the apoptosis of HUVECs in mitochondria dependent manner by enhancing VDAC1 expression[J].Pharmazie, 2018, 73(12):725-728. DOI: 10.1691/ph.2018.8544.
[2] 赵鹏鸣, 王俭勤, 梁耀军. 内皮细胞损伤在糖尿病肾病发病机制中的作用[J]. 中国糖尿病杂志, 2016, 24(2): 169-172. DOI: 10.3969/j.issn.1006-6187.2016.02.020.
[3] 周明杰, 刘立群. 山奈酚预处理对大鼠心脏缺血再灌注损伤的影响[J]. 潍坊医学院学报, 2015,37(3): 168-170. DOI: 10.3969/j.issn.1004-3101.2015.03.003.
[4] 康桂兰, 景增秀. 山奈酚通过调控AMPK/Nrf2/HO-1信号通路缓解ox-LDL介导的内皮细胞损伤[J]. 中国免疫学杂志, 2018, 34(4): 525-530. DOI: 10.3969/j.issn.1000-484X.2018.04.010.
[5] Leung WK, Gao L, Siu PM, et al. Diabetic nephropathy and endothelial dysfunction: Current and future therapies, and emerging of vascular imaging for preclinical renal-kinetic study[J]. Life Sci, 2016,166: 121-130. DOI: 10.1016/j.lfs.2016.10.015.
[6] Wan C, Su H, Zhang C. Role of NADPH Oxidase in Metabolic Disease-Related Renal Injury: An Update[J]. Oxid Med Cell Longev,2016,2016: 7813072.DOI: 10.1155/2016/7813072.
[7] Nagasu H, Satoh M, Kiyokage E, et al. Activation of endothelial NAD(P)H oxidase accelerates early glomerular injury in diabetic mice[J]. Lab Invest, 2016, 96(1): 25-36. DOI: 10.1038/labinvest.2015.128.
[8] 陈娜, 王玉筵, 王秀芬, 等. NADPH氧化酶在肾脏疾病中的研究进展[J]. 中华生物医学工程杂志, 2015, 21(6): 574-577.
[9] Yang Q, Wu FR, Wang JN, et al. Nox4 in renal diseases: An update[J]. Free Radic Biol Med, 2018,124:466-472. DOI: 10.1016/j.freeradbiomed.2018.06.042.
[10] 高攀, 张农. 转录因子NEF2相关因子2在糖尿病肾病中的研究进展[J]. 中国糖尿病杂志, 2015,23(2): 182-184. DOI: 10.3969/j.issn.1006-6187.2015.02.020.
[11] 隆敏, 郑宏庭. 抗氧化的NRF2通路与糖尿病及其慢性并发症[J]. 中华内分泌代谢杂志, 2017, 33(3): 185-189. DOI: 10.3760/cma.j.issn.1000-6699.2017.03.002.
[12] Sifuentes-Franco S, Padilla-Tejeda DE, Carrillo-Ibarra S, et al. Oxidative Stress, Apoptosis, and Mitochondrial Function in Diabetic Nephropathy[J]. Int J Endocrinol, 2018,2018: 1875870. DOI: 10.1155/2018/1875870.
[13] Volpe CMO, Villar-Delfino PH, Dos Anjos PMF, et al. Cellular death, reactive oxygen species (ROS) and diabetic complications[J]. Cell Death Dis, 2018, 9(2):119. DOI: 10.1038/s41419-017-0135-z.
[14] 张微,熊军,王景平,叶红,李莉.缬沙坦联合前列地尔对慢性肾小球肾炎患者肾功能、免疫指标及其细胞因子的影响[J].疑难病杂志,2017,16(12):1236-1239.DOI:10.3969/j.issn.1671-6450.2017.12.013
[15] Sha J, Sui B, Su X, et al. Alteration of oxidative stress and inflammatory cytokines induces apoptosis in diabetic nephropathy[J]. Mol Med Rep, 2017, 16(5):7715-7723. DOI: 10.3892/mmr.2017.7522.
[16] Zhang J, Guo Y, Ge W,et al. High glucose induces the apoptosis of HUVECs in mitochondria dependent manner by enhancing VDAC1 expression[J].Pharmazie, 2018, 73(12):725-728. DOI: 10.1691/ph.2018.8544.