达格列净通过Caspase-1介导的细胞焦亡对糖尿病肾病小鼠肾损伤的影响

Effect of Dapagliflozin on kidney injury in diabetic nephropathy mice through Caspase-1 mediated pyroptosis

目的基于胱天蛋白酶-1(Caspase-1)通路介导的细胞焦亡作用,探究达格列净(DAP)对糖尿病肾病(DN)小鼠肾损伤的影响。方法取30只SPF级C57BL/6雄性小鼠,8周龄,体重17~25 g,根据随机数字表法将其分为对照组、模型组、DAP组,每组10只。模型组、DAP组建立DN模型,对照组给予普通饲料。DAP组灌胃给予1 mg/(kg·d)DAP,对照组及模型组均灌胃给予等量生理盐水,连续给药4周。药物干预后比较各组小鼠的一般情况,血肌酐、24 h尿蛋白、血清白细胞介素-18(IL-18)水平;观察肾脏病理学变化;比较小鼠肾组织Caspase-1通路相关蛋白表达。结果与对照组比较,模型组精神萎靡,活动减少,偶有颤抖,尿量显著增多,体型较瘦;与模型组比较,DAP组有多饮多食现象,尿量增多不明显,活动正常。与对照组比较,模型组24 h尿蛋白、血肌酐、血清IL-18水平及核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)、Caspase-1、GSDMD、IL-18蛋白水平均显著升高(P<0.05);与模型组比较,DAP组小鼠24 h尿蛋白、血肌酐、血清IL-18水平及NLRP3、Caspase-1、GSDMD、IL-18蛋白水平均显著降低(P<0.05)。结论DAP可能通过抑制NLRP3/Caspase-1信号通路活化抑制细胞焦亡,保护肾脏。

Objective To investigate the effect of Dapagliflozin (DAP) on kidney injury in diabetic nephropathy (DN) mice based on the pyrocytosis mediated by cysteine aspartic acid specific protease-1(Caspase-1) pathway. Methods Thirty SPF grade C57BL/6 male mice, aged eight weeks and weighing 17-25 g, were divided into control group, model group, and DAP group with ten mice in each group according to random number table method. DN model was established in model group and DAP group, and ordinary diet was given to control group. DAP group was given 1 mg/ (kg·d) DAP by intragastric administration, while control group and model group were given the same amount of normal saline by intragastric administration for consecutive four weeks. After drug intervention, the general conditions, serum creatinine, 24 h urinary protein, and serum interleuking-18 (IL-18) levels of each group were compared. Pathological changes of kidney were observed. The expression of Caspase-1 pathway related proteins in mouse kidney was compared. Results Compared with the control group, the model group was listless, less active, occasionally shivering, urine volume was increased significantly, and thinner. Compared with model group, DAP group showed polydipsia and hyperbiosis, and the urine volume was not obvious increased and the activity was normal. Compared with control group, 24 h urinary protein, serum creatinine, serum IL-18 levels and NLR family pyrin domain containing 3 (NLRP3), Caspase-1, GSDMD, and IL-18 protein levels in model group were significantly increased (P<0.05). Compared with model group, 24 h urinary protein, serum creatinine and serum IL-18 levels, NLRP3, Caspase-1, GSDMD, and IL-18 protein levels in DAP group were significantly decreased (P<0.05). Conclusion DAP may protect kidney by inhibiting NLRP3/Caspase-1 signaling pathway to activate and inhibit pyrodeath.