三七总皂苷联合氨基胍治疗对糖尿病大鼠肾脏氧化损伤的影响

Effects of combined treatment with panax notoginseng saponins and aminoguanidine on oxidative stress in kidney of diabetic rats

目的 研究三七总皂苷联合氨基胍治疗对糖尿病大鼠肾组织氧化损伤的影响及其机制.方法 清洁级健康雄性SD大鼠48只,腹腔注射链脲佐菌素,建立糖尿病大鼠模型,按数字表法随机分为糖尿病模型组(n=12)、三七总皂苷治疗组(n=12)、氨基胍治疗组(n=12)和三七总皂苷+氨基胍联合治疗组(n=12);另设正常对照组(n=12).第4、8周应用比色法测定血清及肾皮质超氧化物歧化酶及丙二醛含量,采用荧光光谱法测定血清和肾皮质晚期糖基化终产物含量,行肾组织PAS染色并测定肾小球平均截面积,计算肾小球平均体积.采用单因素方差分析进行组间比较.结果 三七总皂苷+氨基胍联合治疗组血清及肾皮质晚期糖基化终产物(F=36.017,P<0.01;F=8.213,P<0.01)、丙二醛(F=8.683,P<0.01;F=14.615,P<0.01)、内生肌酐清除率(F=7.233,P<0.01)、肾小球平均截面积(F=24.317,P<0.01)、肾小球平均体积(F=26.145,P<0.01)、尿蛋白量(F=17.108,P<0.05)、肾脏重量(F=5.182,P<0.05)、肾脏肥大指数(F=50.169,P<0.01)低于糖尿病模型组,血清及肾皮质超氧化物歧化酶(F值分别为6.260、7.666,均P<0.01)、体重(F=10.449,P<0.05)高于糖尿病模型组.三七总皂苷+氨基胍联合治疗组血清丙二醛低于氨基胍治疗组(F=8.683,P<0.05),肾皮质丙二醛低于氨基胍治疗组和三七总皂苷治疗组(F=14.615,P<0.05),血清晚期糖基化终产物低于氨基胍治疗组和三七总皂苷治疗组(F=36.017,P<0.01或<0.05),肾皮质晚期糖基化终产物低于氨基胍治疗组和三七总皂苷治疗组(F=8.213,P<0.05).8周末时,三七总皂苷+氨基胍联合治疗组肾小球平均截面积、肾小球平均体积均低于三七总皂苷治疗组(F值分别为24.317、26.145,均P<0.05)和氨基胍治疗组(F值分别为24.317、26.145,均P<0.05).结论 三七总皂苷+氨基胍联合治疗可通过减轻肾组织氧化应激反应、减少肾组织晚期糖基化终产物的生成与蓄积对糖尿病大鼠肾脏起到保护作用,其作用优于单一药物治疗.

Objective To investigate the effect of combined treatment with panax notoginseng saponins (PNS) and aminoguanidine (AG) on oxidative stress in kidney of diabetic rats as well as possible mechanism. Methods The rat diabetic model was induced by intraperitoneal injection of STZ in the left lower abdomen of the rats. A total of 48 model rats were randomly divided into diabetic group ( n = 12 ), PNS treatment group (n = 12) , AG treatment group (n = 12) , and PNS+AG treatment group (n= 12). Another 12 rats were used as normal controls. Serum and renal cortex superoxide dismutase (SOD) and malondialdehyde (MDA) were measured by colorimetric assay. Advanced glycation end products (AGEs) were determined by spectrum law of fluorescence in the 4 and 8 weeks. The mean glomerular area ( MGA) and mean glomerular volume (MGV) were measured by image analysis system. The changes of the blood glucose levels, urine amount, urine albumen, the renal function, body weight ( BW) , kidney weight (KW) and KW to BW ratio were observed. Results In comparison with the diabetic group, AGEs (F = 36.017, P<0.01;F=8.213, P<0.01) and MDA(F=8.683,P<0.01;F=14.615, P<0.01) in serum and renal cortex, creatinine clearance rate (F = 7.233, P<0.01), MGA(F = 24. 317, P<0.01), MGV(F = 26.145, P<0.01), urinary excretion of protein (F = 17.108, P<0.05), KW( F = 5.182, P<0.05) and KW to BW ratio( F = 50.169, P<0.01 ) were decreased significantly in the PNS + AG treatment group, and SOD(F = 6.260,P<0.01;F=7.666, P<0.01)in serum and renal cortex and BW were increased significantly (F = 10.449,P<0.05). Decreased serum MDA was observed in PNS + AG treatment group compared with AG treatment group ( F = 8.683, P<0.05 ) , although renal cortex MDA was significantly lower than that of PNS treatment group and AG treatment group (F = 14.615, P<0.05). Serum AGEs was decreased significantly in PNS + AG treatment group compared with AG or PNS treatment group ( F = 36.017,P<0.01 or <0.05), and renal cortex AGEs was significantly lower than that of AG or PNS treatment group( F = 8.213, P<0.05). At 8 weeks, lower MGA( F = 24.317 ,P<0.05) and MGV(F = 26.145 ,P<0.05) were found in the PNS + AG treatment group. Conclusion Combined treatment with PNS and AG may possess renoprotective effect on STZ-induced diabetic rats, and the possible mechanism could be associated with reduced oxidative stress and inhibited overproduction of AGEs in renal tissue.