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目的探讨糖尿病大鼠血清及肾组织甘露糖结合凝集素(mannose-binding lectin,MBL1)表达变化及来氟米特治疗后对其表达的影响。方法将56只8周龄清洁级雄性SD大鼠,按随机数字表法分为对照组(n=16)与模型组(n=40);模型组给予链脲佐菌素制作糖尿病大鼠模型,造模成功后再随机分为糖尿病组和来氟米特组,来氟米特组给予来氟米特[5 mg/(kg·d)]治疗性灌胃,对照组及糖尿病组给予等体积生理盐水灌胃,连续8周。分别于成模后4周及8周处死大鼠,检测各组大鼠24 h尿蛋白定量、血糖、血清肌酐、尿素氮,肾脏标本测肾脏质量,PAS染色观察肾脏病理改变。ELISA法检测血清MBL1水平,采用免疫组化、半定量RT-PCR及Western blot检测肾组织MBL1表达。结果与对照组大鼠比较,4、8周糖尿病组及来氟米特组大鼠肾质量指数、血清肌酐、尿素氮、血清MBL1及24 h尿蛋白定量差异均有统计学意义(P<0.01);来氟米特组大鼠肾质量指数、血清肌酐、尿素氮、血清MBL1[4周(1.117±0.067)μg/L vs(1.249±0.054)μg/L,8周(1.222±0.074)μg/L vs(1.470±0.132)μg/L]及24 h尿蛋白定量较糖尿病组大鼠明显降低(尿素氮P<0.05,余指标P<0.01);对照组肾组织MBL1有少量表达,糖尿病组肾组织MBL1表达增加,来氟米特组各时间点与糖尿病组比较均显著降低(P<0.01),相关分析显示血清及肾组织MBL1表达与24 h尿蛋白定量、血清肌酐、尿素氮、肾质量指数、平均肾小球横截面积、平均肾小球体积呈显著正相关。结论 MBL1参与糖尿病肾病发生、发展,来氟米特可以通过减低血清及肾组织MBL1的水平,降低炎症反应,减轻糖尿病大鼠肾脏损害。
Objective To investigate the expression of mannose-binding lectin (MBL1) in serum and kidney of diabetic rats and the effect of leflunomide on the expression of MBL1. Methods 56 male SD rats of 8 weeks of age were divided into control group (n = 16) and model group (n = 40) according to random number table. The model group was given streptozotocin to make diabetic rat model , And then were randomly divided into diabetic group and leflunomide group after successful modeling. Leflunomide group was given leflunomide [5 mg / (kg · d)] intragastrically, and the control group and diabetic group were given The volume of normal saline gavage for 8 weeks. The rats were sacrificed at 4 weeks and 8 weeks after operation respectively. 24 h urinary protein, blood glucose, serum creatinine, blood urea nitrogen and kidney were measured 24 hrs after operation. The renal pathological changes were observed by PAS staining. Serum MBL1 levels were detected by ELISA. Immunohistochemistry, semi-quantitative RT-PCR and Western blot were used to detect the expression of MBL1 in renal tissues. Results Compared with the control group, the quantitative difference of renal mass index, serum creatinine, urea nitrogen, serum MBL1 and 24 h urinary protein in diabetic group and leflunomide group at 4 and 8 weeks were statistically significant (P <0.01 (1.117 ± 0.067) μg / L vs (1.249 ± 0.054) μg / L, 8 weeks (1.222 ± 0.074) μg / L for leflunomide group, serum creatinine, / L vs (1.470 ± 0.132) μg / L] and 24 h urinary protein in diabetic rats were significantly lower than those in diabetic rats (urea nitrogen, P <0.05, remaining indicators, P <0.01) The expression of MBL1 in renal tissue increased, while in leflunomide group was significantly lower than that in diabetic group at all time points (P <0.01). Correlation analysis showed that the expression of MBL1 in serum and kidney tissue was correlated with 24 h urinary protein, serum creatinine, urea nitrogen, Quality index, the average glomerular cross-sectional area, the average glomerular volume was significantly positively correlated. Conclusions MBL1 is involved in the development and progression of diabetic nephropathy. Leflunomide can reduce the level of MBL1 in serum and renal tissue, reduce the inflammatory reaction and reduce the renal damage in diabetic rats.