金芪降糖片对糖尿病大鼠肾脏IL-17和IL-23表达的影响

The effect of Jinqi Jiangtang tablet on expressions of IL-17 and IL-23 in kidney of diabetic rats

目的探讨金芪降糖片对糖尿病大鼠辅助性T细胞(Th)17以及肾脏白细胞介素(IL)-17和IL-23表达的影响。方法雄性SD大鼠45只,随机分为正常对照组(NC组,15只)和造模组(30只),造模组通过一次性尾静脉注射链脲佐菌素(STZ,45 mg/kg)诱导建立糖尿病模型,造模成功的28只大鼠随机分为糖尿病组(DM组)和金芪降糖片治疗组(金芪组,2.1 g·kg^(-1)·d^(-1)),各14只。金芪组给予每日1次金芪降糖片溶液灌胃18周,NC组和DM组给予等体积生理盐水灌胃。18周末取材。流式细胞术测定外周血Th17表达频率;双抗体两步夹心酶联免疫吸附法(ELISA)测定血清IL-17和IL-23水平;电镜观察各组大鼠肾小球超微结构改变;免疫组化法测定肾脏IL-17和IL-23的表达水平。结果 (1)DM组和金芪组大鼠外周血Th17/CD4^+细胞比例均较NC组显著上升,金芪组大鼠外周血Th17/CD4^+细胞比例较DM组降低。(2)DM组和金芪组大鼠血清IL-17、IL-23水平均较NC组显著上升,金芪组IL-17、IL-23水平较DM组降低。(3)电镜下,DM组大鼠肾小球毛细血管基底膜增厚,部分足细胞突起融合,系膜细胞增生;金芪组大鼠肾小球病变较DM组减轻。(4)DM组和金芪组大鼠肾脏IL-17、IL-23表达均较NC组显著上升,金芪组IL-17、IL-23表达均较DM组降低。结论 Th17活化、肾脏IL-17和IL-23表达增多可能在糖尿病肾病发病过程中起重要作用。金芪降糖片可能通过抑制Th17活化和减少肾脏IL-17和IL-23的表达来改善糖尿病肾病。

Objective To investigate the effect of Jinqi Jiangtang tablet on the activation of T helper type 17 (Th17) and the expressions of interleukin (IL)-17 and IL-23 in kidney of diabetic rats. Methods A total of 45 male SD rats were randomly divided into normal control group (NC, n=15) and experimental group (n=30). Diabetes was induced by tail vein injection with streptozotocin (STZ, 45 mg/kg). The well-established 28 diabetic model rats were then randomly divided into diabetes group (DM, n=14) and Jinqi Jiangtang tablet administration group (Jinqi, n=14). The rats in Jinqi group were given Jinqi Jiangtang tablet solution by gavage at a single dose of 2.1 g·kg-1·d-1 for 18 weeks, while NC group and DM group were given 0.9% NaCl in the same way. All rats were sacrificed after 18 weeks. The circulating Th17 frequencies were assessed using flow cytometry. Serum IL- 17 and IL- 23 levels were measured by enzyme- linked immunosorbent assay. The pathological changes in kidney were studied by electron microscope. The expressions of IL-17 and IL-23 in kidney were detected using immunohistochemistry. Results (1) Compared with the group NC, the circulating Th17 frequencies were significantly increased in group DM and group Jinqi. The circulating Th17 frequencies were significantly lower in group Jinqi than those in group DM. (2) Compared with the group NC, the serum IL-17 and IL-23 levels were significantly increased in group DM and group Jinqi. The serum IL-17 and IL-23 levels were significantly lower in group Jinqi than those in group DM. (3) In group DM, irregular thickening of glomerular basement membrane, fusion of epithelial cell foot processes and mesangial expansion were observed by electron microscope. The above-mentioned pathological changes were improved in the group Jinqi. (4) Compared with the group NC, the expressions of IL-17 and IL-23 in the renal cortex were significantly increased in group DM and group Jinqi, and those were significantly lower in group Jinqi than those in group DM.Conclusion The activation of Th17 and the increased expressions of IL-17 and IL-23 in kidney play a potential role in diabetic nephropathy. Jinqi Jiangtang tablet can improve diabetic nephropathy through inhibiting the activating Th17 and decreasing the expression of IL-17 and IL-23 in kidney.