摘要
目的观察1,25(OH)_2D_3对肾小球系膜细胞(HMC)凋亡相关因子表达的影响。方法2017年10月—2018年9月在武汉科技大学附属普仁医院肾内科实验室经体外培养HMC,并按随机数字表法分成4组:A组(空白对照组)、B组(EGF组)、C组[1,25(OH)_2D_3组]、D组[EGF、1,25(OH)_2D_3联合组]。检测Caspase蛋白活性,Western bolt法测定Akt、p-Akt及Caspase-3、Caspase-8、Caspase-9蛋白表达,选择荧光实时定量PCR法测定Caspase-3,Caspase-8、Caspase-9 mRNA表达。结果各组Caspase-3蛋白活性表达比较,B组<A组<D组<C组(q/P=5.125/<0.001、9.418/<0.001、7.263/<0.001);p-Akt/Akt比值C组<A组<D组<B组(q/P=8.263/<0.001、13.572/<0.001、17.424/<0.001);Caspase-3、Caspase-9蛋白表达C组>D组> A组>B组(q/P=5. 156/<0.001、5.436/<0.001、7. 326/<0.001;5.074/<0. 001、5. 982/<0. 001、6. 513/<0. 001); Caslpase-8蛋白的表达4组比较差异无统计学意义(P>0. 05);Caspase-3 mRNA及Caspase-8 mRNA基因表达C组> A组> D组> B组,Caspase-9 mRNA表达C组> A组> B组> D组(q/P值=15. 672/<0. 001、17.526/<0. 001、19. 421/<0. 001;5. 168/<0. 001、5.689/<0.001、6.024/<0.001;6.425/<0.001、6.974/<0. 001、7. 654/<0. 001)。结论 1,25(OH)_2D_3通过增强HMC活性,抑制Akt磷酸化进程,提高Caspase-3及Caspase-9 mRNA表达水平,进而加速促进HMC凋亡。
Objective To observe the effect of 1,25(OH)2 D3 on the expression of apoptosis-related factors in glomerular mesangial cells( HMC). Methods From October 2017 to September 2018, HMC was cultured in vitro in the Laboratory of Renal Medicine, Puren Hospital Affiliated to Wuhan University of Science and Technology. They were randomly divided into four groups: group A( blank control group), group B( EGF group), group C [ 1, 25(OH)2 D3 group] and group D [EGF, 1, 25(OH)2 D3 combined group]. The activity of Caspase was detected. The expression of Akt, p-Akt, Caspase-3, Caspase-8 and Caspase-9 protein was determined by Western bolt method. The expression of Caspase-3, Caspase-8 and Caspase-9 gene was determined by real-time fluorescence quantitative PCR. Results Caspase 3 activity was expressed in group B < group A < group D < group C(q/P =5. 125/< 0. 001,q/P =9. 418/<0. 001,q/P =7. 263/<0. 001);p-Akt/Akt ratio Group C < Group A < Group D < Group B(q/P = 8. 263/<0. 001,q/P = 13. 572/<0. 001,q/P =17.424/<0.001);Caspase-3, Caspase-9 protein expression group C > Goup D > Group A > group B(q/P =5. 156/<0.001,q/P=5. 436/<0. 001,q/P =7. 326/< 0. 001,q/P =5. 074/<0.001,q/P =5. 982/< 0. 001,q/P =6. 513/<0.001). There was no significant difference in the expression of Caspase-8 protein between the 4 groups( P > 0. 05).Caspase-3 mRNA and Caspase-8 mRNA gene expression group C > group A > group D > group B, Caspase-9 mRNA expression group C > group A > group B > group D(q/P = 15.672/<0.001,q/P = 17.526/<0.001,g/P = 19.421/<0.001,q/P =5. 168/<0.001,q/P=5.689/<0.001,q/P = 6.024/<0.001,q/P = 6.425/<0.001,q/P = 6. 974/<0.001,q/P=7.654/<0.001). Conclusion 1,25(OH)2 D3 can enhance the activity of HMC, inhibit the phosphorylation of Akt, increase the expression of Caspase-3 and Caspase-9, and then accelerate the apoptosis of HMC.
作者
唐小铁
徐洁
王丽媛
于洋
崔学彬
孙维言
TANG Xiaotie;XU Jie;WANG Liyuan;YU Yang;CUI Xuebin;SUN Weiyan(Department of Nephrology,Puren Hospital Affiliated to Wuhan University of Science and Technology,Hubei Province,Wuhan 430081,China)
出处
《疑难病杂志》
CAS
2019年第5期503-506,共4页
Chinese Journal of Difficult and Complicated Cases
基金
湖北省卫计委联合基金面上项目(WJ2018H0107)
武汉市卫计委一般项目(WX17D27)
