补肾活血方药对糖尿病肾病大鼠肾脏中晚期糖基化终末产物的影响及可能机制

Effects and possible Mechanism of Bushen- huoxue formulas on AGEs in kidney of diabetic rats

目的探讨补肾活血方药对糖尿病肾病大鼠肾脏晚期糖基化终末产物(AGEs)的影响及可能机制。方法将48只雄性SD大鼠随机分为4组,即正常对照组、模型组、氨基胍组及补肾活血方组,除正常对照组外,其余3组大鼠予链脲佐菌素腹腔注射建立糖尿病肾病大鼠模型。造模成功后,补肾活血方组大鼠予6.3 g生药/kg补肾活血方水煎剂,氨基胍组大鼠予含盐酸氨基胍的自来水50 mg/(kg·d),模型组和正常对照组大鼠予等容积自来水灌胃,均每日1次,连续8周。测定各组大鼠空腹血糖(FBG)、肾质量/体质量、血尿素氮(BUN)、血肌酐(Cr)、24 h尿蛋白定量,检测肾小球AGEs(GTE–AGEs)及血清和尿中抗-羧甲基赖氨酸(CML)-AGEs、荧光性-AGEs含量,生物化学法测定肾小球脂质过氧化物(LPO)含量,观察肾脏病理改变。结果与正常对照组相比,模型组大鼠FBG、BUN、Cr、肾质量/体质量、24 h尿蛋白定量、GTE-AGEs、血清荧光性-AGEs及CML-AGEs、肾小球LPO含量均显著升高(P<0.05,P<0.01);与模型组比较,补肾活血方组及氨基胍组FBG、BUN、Cr、肾质量/体质量、24 h尿蛋白定量、GTE-AGEs、肾小球LPO显著降低,尿荧光性-AGEs、尿CML-AGEs含量显著升高(P<0.05)。与正常对照组比较,模型组肾小球平均截面积(MGA)、肾小球平均体积(MGV)显著增大(P<0.01);与模型组比较,补肾活血方组及氨基胍组MGA、MGV显著减小(P<0.05,P<0.01),2组肾脏病理学改变显著减轻,且2组组间比较差异无统计学意义(P>0.05)。结论补肾活血方可通过增加尿液AGEs排泄减少AGEs在糖尿病肾病大鼠肾脏的蓄积,降低尿蛋白排泄,减少氧化应激水平,从而发挥其对糖尿病肾病的防治作用。

Objective Explore the effects and possible mechanism of Bushen - huoxue formulas on advanced glycosylation end products （AGEs） in kidneys in diabetic rats. Methods 48 male SD rats were randomly divided into four groups ： normal control group （ group N ）, model group （ group DM ）, positive control medicine group with aminoguanidine （ group AG） , and treatment group with Bushen - huoxue formulas （ group BSHX）. Diabetes models were induced by injection of streptozocin. After the treatment of aminoguanidine, Bushen - huoxue for 8 weeks, kidney weight/body weight, blood urea nitrogen （BUN）, serum creatinine （Cr）, total 24 h urine protein were measured for each group; glomerular AGEs （GTE - AGEs）, serum and urine of CML - AGEs. fluorescent - AGEs were detected with enzyme - linked immunoassay and fluorescence; The content of glomerular lipid peroxide （LPO） was detected with biochemical method. The pathological changes of kidney were observed. Result The level of FBG, BUN, Cr, kidney weight/body weight, 24 h urine protein, GTE - AGEs, serum fluorescence - AGEs, the serum levels of CML - AGEs and GTE - LPO were all significantly higher in group DM than those in group N （ P 〈 0.05, P 〈0.01 ）. The FBG, BUN, Cr, kidney weight/bodyweight, 24h urine protein, GTE - AGEs and GTE - LPO were significantly lower in group BSHX and group AG than those in group DM; the content of urine fluorescence - AGEs, urine CML - AGEs were significantly higher. MGA and MGV in group DM were all significantly increased as compared with those in group N （P 〈 0.01 ）. MGA and MGV in group BSHX and group AG were all significantly increased as compared with those in group DM （ P 〈 0.05, P 〈 0.01 ）. The pathological changes of kidney were reduced in the rats of group BSHX and group AG, there was no significant difference between group BSHX and group AG on renal pathology change （P 〉 0. 05 ）. Conclusion Bushen - huoxne formulas can reduce accumulation of AGEs in the kidney of diabetic rats, reduce the excretion of urinary protein, and reduce oxidative stress levels by in- creasing the excretion of urine AGEs. Thus, it plays roles in prevention and treatment of diabetic nephropathy.