黄芪多糖对高糖刺激小鼠肾小球足细胞Nephrin和Desmin表达的影响
The Effect of Astragalus Polysaccharide on the Expressions of Nephrin and Desmin in High Sugar Stimulation Mice Podocytes
-
摘要: 目的 模拟体外高糖病理微环境, 探讨黄芪多糖对实验鼠肾脏肾小球足细胞Nephrin及Desmin表达的影响.方法 试验 (1) :将培养的实验鼠肾脏肾小球足细胞分为5个组, 分别是D-葡萄糖30 mmol/L+黄芪多糖干预组 (0.0 g/L, 0.1 g/L, 0.2 g/L, 0.4 g/L及0.8 g/L) ;干预时间为72 h, 筛选出黄芪多糖干预的最佳浓度点0.Z g/L.试验 (2) :将培养的实验鼠肾脏肾小球的足细胞分7个组, 分别是D-葡萄糖30 mmol/L+黄芪多糖的干预 (干预浓度是0.Z g/L) , 干预的时间分别是0 h, 3 h, 6 h, 12 h, 24 h, 48 h及72 h;筛选出干预的最佳时间点Y h.试验 (3) :根据上述试验 (1) 、 (2) 结果, 将培养的小鼠肾小球足细胞分为:正常对照组、甘露醇高渗对照组、高糖组和黄芪多糖干预组 (黄芪多糖的干预浓度是0.Z g/L, 干预的时间是Y h) , 采用流式细胞术与实时聚合酶链反应 (real-Time Polymerase Chain Reaction, real-Time PCR) 分别检测七组足细胞Nephrin、Desmin蛋白与m RNA的表达.结果 随着黄芪多糖干预梯度浓度的递增, 实验鼠肾脏肾小球的足细胞Nephrin蛋白表达逐渐上调, Desmin蛋白表达逐渐下调, 呈浓度依赖性 (P<0.05) ;随着黄芪多糖干预时间的延长, 实验鼠肾脏肾小球的足细胞Nephrin的表达逐渐上调, 但肾小球的足细胞Desmin的表达逐渐下调, 呈时间依赖性.相比正常对照组和甘露醇高渗对照组, 高糖组小鼠肾小球足细胞Nephrin蛋白和m RNA表达均下降, Desmin蛋白和m RNA表达均增加 (P<<0.05) ;而相较高糖组, 黄芪多糖干预组Nephrin蛋白和m RNA表达明显上升, Desmin蛋白和m RNA表达明显降低 (P<0.05) .结论 在高糖刺激下, 黄芪多糖干预可上调肾小球足细胞Nephrin表达, 降低肾小球足细胞Desmin表达, 是黄芪治疗DN蛋白尿的可能作用靶点.Abstract: Objective To study the effect of Astragalus polysaccharide on the expressions of Nephrin and Desmin in high sugar stimulation mice podocytes. Me thods Test one: the cultured mouse podocytes were divided into 5 groups: astragalus polysaccharide group [D-glucose 30 mmol/L + astragalus polysaccharide (0.0 g/L, 0.1 g/L, 0.2 g/L, 0.4 g/L and 0.8 g/L) ]. Test two: In test one, we chose the concentration of astragalus polysaccharide which most affected both expression of Nephrin and Desmin in podocytes and named it as astragalus polysaccharide 0.X g/L. The cultured mouse podocytes were divided into 7 groups: astragalus polysaccharide group (D-glucose 30 mmol/L + astragalus polysaccharide 0.X g/L) had been cultured for 0 h, 3 h, 6 h, 12 h, 24 h, 48 h and 72 h, respectively. Test three: Basted on test two, we chose the time which most affected both expression of Nephrinand Desmin in podocytes and named it as Yh. The cultured mouse podocytes were divided into 4 groups: the control group, mannitol hypertonic glucose control group, high sugar group, and astragalus polysaccharide group (high sugar group + astragalus polysaccharide 0.X g/L) . Each group had been cultured for Yh. Nephrin and Desmin of each subgroup was detected by flow cytometry podocytes and real-time PCR. Re s ults With the intervention of polysaccharide, Nephrin in podocyte cell surface gradually increased, Desmin gradually reduced in a concentration-dependent manner ( P <0.05) . With the intervention of astragalus polysaccharide, Nephrin gradually increase and Desmin expression gradually decreased in a concentration-dependent manner (P<0.05) .Compared to the control group and the high sugar group, Nephrin mR NA increased and Desmin mR NA decreased in0.8 g/L astragalus polysaccharide co-cultured for 72 h ( P <0.05) . Conclus ion Astragalus polysaccharide intervention can increase Nephrin and reduce Desmin in podocytes, which are possible targets of Astragalus polysaccharide in treatment of DN proteinuria.
-
Key words:
- Astragalus polysaccharide /
- Podocytes /
- Nephrin /
- Desmin
-
[1] [1]LI M, WANG W, XUE J, et al.Meta-analysis of the clinical value of Astragalus membranaceus in diabetic nephropathy[J].J Ethnopharmacol, 2011, 133 (1) :412-419. [2] [2]BOSE M, ALMAS S, PRABHAKAR S.Wnt signaling and podocyte dysfunction in diabetic nephropathy[J].J Investig Med, 2017, 65 (8) :1093-1101. [3] [3] GUO C, LIU Y, ZHAO W, et al.Apelin promotes diabetic nephropathy by inducing podocyte dysfunction via inhibiting proteasome activities[J].J Cell Mol Med, 2015, 19 (9) :2273-2285. [4] [4]NAKHJAVANI M, AGHAJANI NARGESI A, SALABATI M, et al.Changes in Leukocyte Subpopulations with Decline in Glomerular Filtration Rate in Patients with Type 2Diabetes[J].Acta Med Iran, 2015, 53 (7) :425-431. [5] [5]JOURDAN T, SZANDA G, ROSENBERG A Z, et al.Overactive cannabinoid 1 receptor in podocytes drives type2 diabetic nephropathy[J].Proc Natl Acad Sci U S A, 2014, 111 (50) :E5420-E5428. [6] [6]DUMONT V, TOLVANEN TA, KUUSELA S, et al.PACSIN2 accelerates nephrin trafficking and is up-regulated in diabetic kidney disease[J].FASEB J, 2017, 31 (9) :3978-3990. [7]李志杰, 张悦, 刘煜敏, 等.黄芪多糖对早期糖尿病肾病大鼠足细胞nephrin和podocin表达的影响[J].中国病理生理杂志, 2011, 27 (09) :1772-1776. [8] [8]HONG T, CUI LK, WEN J, et al.Cordycepin protects podocytes from injury mediated by complements complex C5b-9[J].Sichuan Da Xue Xue Bao Yi Xue Ban, 2015, 46 (2) :173-227. [9] [9]ZHAI L, GU J, YANG D, et al.Metformin ameliorates podocyte damage by restoring renal tissue nephrin expression in type 2 diabetic rats[J].J Diabetes, 2017, 9 (5) :510-517. [10] [10]GUO C, LIU Y, ZHAO W, et al.Apelin promotes diabetic nephropathy by inducing podocyte dysfunction via inhibiting proteasome activities[J].J Cell Mol Med, 2015, 19 (9) :2273-2285. [11] [11]ZHANG J, HU X, WANG S, et al.Protective effects of low-dose rapamycin combined with valsartan on podocytes of diabetic rats[J].Int J Clin Exp Med, 2015, 8 (8) :13275-13281. [12]王竹, 刘俊田, 孙万森, 等.祛风通络方对阿霉素肾病大鼠足细胞Desmin及CD2AP蛋白的影响[J].中国中西医结合杂志, 2014, 34 (2) :203-208. [13]张智颖.解毒通络保肾胶囊对实验性糖尿病肾病大鼠Desmin的实验研究[D].长春中医药大学, 2013. [14] [14]XIAO F, HU Y G, WU S N, et al.Protective effect of astragalus saponin extracts on kidneys of diabetic rats[J].Zhongguo Zhong Yao Za Zhi, 2015, 40 (10) :2014-8. [15] [15]ZHANG J, XIE X, LI C, et al.Systematic reviewof the renal protective effect of Astragalus membranaceus (root) on diabetic nephropathy in animal models[J].J Ethnopharmacol, 2009, 126 (2) :189-196. -
点击查看大图
计量
- 文章访问数: 1560
- HTML全文浏览量: 577
- PDF下载量: 174
- 被引次数: 0
下载:
