[1]张 磊,刘超楠,韩秀涛,等.雷公藤乙素对糖尿病肾病足细胞损伤自噬作用及免疫代谢的影响[J].陕西中医,2023,(8):1021-1026.[doi:DOI:10.3969/j.issn.1000-7369.2023.08.006]
　ZHANG Lei,LIU Chaonan,HAN Xiutao,et al.Effect of triptolide on autophagy and immunity of DKD podocyte injury[J].,2023,(8):1021-1026.[doi:DOI:10.3969/j.issn.1000-7369.2023.08.006]
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雷公藤乙素对糖尿病肾病足细胞损伤自噬作用及免疫代谢的影响
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《陕西中医》[ISSN:1000-7369/CN:61-1281/TN]

卷:
期数:
2023年8期
页码:
1021-1026
栏目:
基础研究
出版日期:
2023-08-05

文章信息/Info

Title:
Effect of triptolide on autophagy and immunity of DKD podocyte injury
作者:
张 磊1; 2; 3; 刘超楠1; 2; 3; 韩秀涛1; 2; 赵静宇1; 2; 周晓霜1; 2
(1.山西中医药大学,山西 晋中 030619; 2.山西省人民医院,山西 太原 030012; 3.首都医科大学附属北京佑安医院 北京肝病研究所,北京 100069)
Author(s):
ZHANG Lei; LIU Chaonan; HAN Xiutao; ZHAO Jingyu; ZHOU Xiaoshuang
(Shanxi University of Traditional Chinese Medicine,Jinzhong 030619,China)
关键词:
糖尿病肾病;  雷公藤乙素;  足细胞;  树突状细胞;  T淋巴细胞;  糖代谢
Keywords:
Diabetic kidney disease;  Triptolide;  Podocyte;  Dendritic cell;  T lymphocytes;  Glucose metabolism
分类号:
R 256.5
DOI:
DOI:10.3969/j.issn.1000-7369.2023.08.006
文献标志码:
A
摘要:
目的:探讨雷公藤乙素对糖尿病肾病(DKD)足细胞损伤自噬作用及DKD患者树突状细胞、T淋巴细胞、糖代谢的影响。方法:以体外培养小鼠足细胞系为研究对象,将足细胞分为正常对照组、高糖组和高糖+雷公藤乙素组。Western blot检测三组足细胞特异性指标肾病蛋白(Nephrin)、肾小球足细胞裂隙膜蛋白(Podocin)及自噬相关蛋白LC3、p62、哺乳动物雷帕霉素靶蛋白(mTOR)表达情况; RT-PCR法检测三组足细胞炎症因子白介素-1α(IL-1α)、白介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)mRNA表达水平。收集健康者和DKD患者外周静脉血单核细胞向树突状细胞(DC)分化,混合淋巴细胞反应。将其分为正常对照组、DKD组和DKD+雷公藤乙素组。流式细胞仪检测DC表面分子HLA-DR MHCⅡ、CD86、CD54水平,T淋巴细胞CD3+、CD4+、CD8+水平以及用HLA-DR MHCⅡ标记的DC细胞葡萄糖转运蛋白-1(GLUT-1)水平。结果:与高糖组比较,高糖+雷公藤乙素组Nephrin、mTOR蛋白表达升高,p62表达降低(均P<0.05),LC3、Podocin蛋白表达虽升高,但差异无统计学意义(均P>0.05)。与高糖组比较,高糖+雷公藤乙素组IL-1α、TNF-α mRNA表达水平明显改善(均P<0.05),IL-1β、IL-6 mRNA表达水平虽有改善,但是差异无统计学意义(均P>0.05)。与DKD组比较,DKD+雷公藤乙素组DC表达HLA-DR MHCⅡ、CD86、CD54平均荧光值较高(均P<0.05)。与DKD组比较,DKD+雷公藤乙素组T淋巴细胞CD3+、CD4+和CD8+表达升高(均P<0.05)。与DKD组比较,DKD+雷公藤乙素组GLUT-1表达量升高(P<0.05)。结论:雷公藤乙素能够减轻足细胞损伤,激活自噬的活性,能够使DC有效激活T淋巴细胞,改善免疫功能,并能够调节糖代谢。
Abstract:
Objective:To investigate the effect of triptolide on podocyte injury and autophagy in diabetic kidney disease(DKD),and the effects of triptolide on dendritic cells,T lymphocytes and glucose metabolism in DKD patients.Methods:Mouse podocyte cultured in vitro were divided into normal control group,high glucose group and high glucose+triptolide group.Western blot was used to detect the expression of podocyte specific indicators,Nephrin,Podocin and autophagy related proteins LC3,p62,mTOR.The mRNA expression levels of inflammatory cytokines interleukin-1α(IL-1α),interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in the three groups were detected by RT-PCR.Collect peripheral venous blood monocytes from healthy people and DKD patients to differentiate into DC,and mix lymphocyte reaction,and they were divided into normal control group,DKD group,and DKD+triptolide group.Flow cytometry was used to detect the expression of HLA-DR MHC Ⅱ,CD86,and CD54 on the surface of DC cells,as well as the expression of CD3+,CD4+,and CD8+ on T lymphocytes,as well as the expression of glucose transporter-1(GLUT-1)in DC cells labeled with HLA-DR MHC Ⅱ.Results:Compared with the high glucose group,the protein expressions of Nephrin and mTOR in the high glucose+triptolide group were increased,while the expression of p62 was decreased(all P<0.05); The protein expressions of LC3 and Podocin were increased,but the differences were not statistically significant(all P>0.05).Compared with high glucose group,mRNA expression levels of IL-1α and TNF-α in high glucose+triptolide group were significantly improved(all P<0.05),the mRNA expression levels of IL-1β and IL-6 were improved,but the differences were not statistically significant(all P>0.05).Compared with DKD group,DKD+triptolide group expressed higher mean fluorescence values of MHCⅡ,CD86,CD54(all P<0.05).Compared with DKD group,expressions of CD3+,CD4+ and CD8+ in T lymphocytes of DKD+triptolide group were increased(all P<0.05).Compared with DKD group,GLUT-1 expression in DKD+triptolide group increased(P<0.05).Conclusion:Triptolide can alleviate podocyte damage,activate autophagy activity,effectively activate T lymphocytes through dendritic cells,improve immune function,and regulate glucose metabolism.

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备注/Memo

备注/Memo:
基金项目:山西省科技厅基础研究计划项目(202103021222013)
更新日期/Last Update: 2023-08-10