【摘要】 目的：探究大黄素通过调节整合素连接激酶(ILK)/丝裂原活化蛋白激酶(MAPK)信号通路对糖尿病肾脏病大鼠肾组织的改善作用。方法：采用腹腔注射60 mg/kg链脲佐菌素的方法诱导糖尿病肾脏病(DN)大鼠模型，并按随机数字表法分为模型组、大黄素低剂量组、大黄素中剂量组、大黄素高剂量组、二甲双胍组，每组12只。另取12只SD大鼠腹腔注射等量生理盐水，设为假手术组。经药物治疗后，检测大鼠24 h尿蛋白定量、血清肌酐(Scr)水平、肾组织病理形态改变、血清白细胞介素-17(IL-17)和诱导型一氧化氮合酶(iNOS)水平、肾组织ILK/MAPK通路相关蛋白表达。体外培养人肾小管上皮细胞HK-2，随机分为对照组、模型组、大黄素组、ILK过表达质粒组、ILK空载质粒组、大黄素+ILK过表达质粒组。除对照组外，其余各组均以含有30 mmol/L葡萄糖的高糖培养液诱导构建高糖损伤细胞模型，同时分别以药物及质粒处理后，检测各组细胞活力、释放炎性介质IL-17、iNOS水平、ILK/MAPK通路蛋白表达。结果：与假手术组比较，模型组大鼠肾组织呈现明显病理损伤，24 h尿蛋白定量、Scr、血清IL-17及iNOS水平、肾组织p-p38 MAPK/p38 MAPK、ILK蛋白表达均明显升高(P<0.05)；与模型组比较，大黄素低、中、高剂量组和二甲双胍组大鼠肾组织病理损伤均减轻，24 h尿蛋白定量、Scr、血清IL-17及iNOS水平、肾组织p-p38 MAPK/p38 MAPK、ILK蛋白表达均明显降低，且大黄素各剂量组之间呈剂量依赖性(P<0.05)；大黄素高剂量组各指标与二甲双胍组比较，差异无统计学意义(P>0.05)。与对照组比较，模型组HK-2细胞活力明显降低(P<0.05),IL-17与iNOS水平、细胞p-p38 MAPK/p38 MAPK、ILK蛋白表达均明显升高(P<0.05)。与模型组、ILK空载质粒组、大黄素+ILK过表达质粒组比较，大黄素组HK-2细胞活力均明显升高(P<0.05),IL-17与iNOS水平、细胞p-p38 MAPK/p38 MAPK、ILK蛋白表达均降低(P<0.05)，而ILK过表达质粒组细胞活力下降，IL-17与iNOS水平、细胞p-p38 MAPK/p38 MAPK、ILK蛋白表达均升高(P<0.05)。结论：大黄素可通过抑制ILK/MAPK信号，阻碍炎症产生及进展，减轻高糖引发的肾小管上皮细胞损伤，大黄素还可通过抑制炎症缓解肾组织损伤，改善DN大鼠肾功能。更多还原

【Abstract】 Objective: To explore the improving effect of emodin on the renal tissue of diabetic nephropathy rats by regulating the integrin-linked kinase(ILK)/mitogen-activated protein kinase(MAPK) signaling pathway. Methods:The rat model of diabetes nephropathy(DN) was induced by intraperitoneal injection of 60 mg/kg streptozotocin,and was divided into model group, emodin low dose group, emodin medium dose group, emodin high dose group and metformin group according to the random number table, with 12 rats in each group. Another 12 SD rats were intraperitoneally injected with the same amount of normal saline and set as the sham operation group. After drug treatment, the 24 hour urine protein quantitation, serum creatinine(Scr) level, the pathological and morphological changes of renal tissue, the serum inflammatory mediator interleukin-17(IL-17) and inducible nitric oxide synthase(iNOS) levels, and the expression of ILK/MAPK pathway related proteins of renal tissue in rat were detected. Human renal tubular epithelial cells HK-2 were cultured in vitro and randomly divided into control group, model group,emodin group, ILK overexpression plasmid group, ILK empty plasmid group, and emodin+ILK overexpression plasmid group. Except for the control group, the other groups were induced to construct a high-glucose injury cell models by a high-glucose culture medium containing 30 mmol/L glucose. At the same time, after treatment with drugs and plasmids, the cell viability, release of inflammatory mediators IL-17, iNOS level and expression of ILK/MAPK pathway protein were detected in each group. Results: Compared with the sham operation group, the renal tissue of the model group showed obvious pathological damage, and the 24 hour urine protein content, Scr,serum IL-17 and iNOS levels, the protein expression of p-p38 MAPK/p38 MAPK and ILK in renal tissue were significantly increased(P<0.05). Compared with the model group, the pathological damage of the renal tissue of the rats in the low, medium and high dose groups of emodin and metformin groups were reduced, the 24 hour urine protein content, Scr, serum IL-17 and iNOS levels, the protein expression of p-p38 MAPK/p38 MAPK and ILK in renal tissue were significantly decreased, and there was a dose-dependent relationship between the various dose groups of emodin(P<0.05). Compared with metformin group, there was no significant difference in the indexes of high-dose emodin group(P>0.05). Compared with the control group, the viability of HK-2 cells in the model group was significantly decreased(P<0.05), and the levels of IL-17 and iNOS, the protein expression of p-p38MAPK/p38 MAPK and ILK were significantly increased(P<0.05). Compared with the model group, the ILK empty plasmid group and the emodin+ILK overexpression plasmid group, the viability of HK-2 cells in emodin group was significantly increased(P<0.05), and the level of IL-17 and iNOS, the protein expression of cell p-p38 MAPK/p38 MAPK, and ILK were all decreased(P<0.05), while the cell viability was decreased, and the level of IL-17 and iNOS, the protein expression of p-p38 MAPK/p38 MAPK, and ILK in cell were increased in ILK overexpression plasmid group(P<0.05). Conclusion: Emodin can inhibit ILK/MAPK signaling to prevent the occurrence and progression of inflammation, reduce the damage of renal tubular epithelial cells caused by high glucose. Emodin can also alleviate renal tissue damage and improve renal function in DN rats by inhibiting inflammation.更多还原