川芎嗪对链脲佐菌素诱导糖尿病肾病的治疗意义

Therapeutical Significance of Ligustrazine for Treating Streptozotocin- Induced Diabetic Nephropathy

目的研究川芎嗪对糖尿病肾病的治疗意义。方法选取Wistar大鼠30只,随机取10只作为正常对照组(A组),其余20只腹腔注射链脲佐菌素(STZ)60 mg/kg诱发糖尿病后,均分为2组,川芎嗪治疗组(C组)给予川芎嗪100 mg/(kg·d),糖尿病对照组(B组)和A组每天用等容量0.9%氯化钠注射液每100 g体重1 m L,分别于第4,8,12,16周测定血糖和留取24 h尿液测定尿白蛋白定量。16周后处死大鼠,分离肾脏,测定组织醛糖还原酶活性,用免疫组化法观察Bcl-2及Bax蛋白的表达,取部分肾皮质病理观察和电镜观察肾脏细胞凋亡形态学改变。结果 C组的Bcl-2蛋白表达较B组增多,而Bax蛋白表达减少;C组醛糖还原酶(AR)活性较B组显著下降(P=0.000),虽较A组高但差异无统计学意义(P>0.05)。B组血糖、醛糖还原酶活性均显著高于A组(P<0.01);C组电镜下凋亡形态学改变不明显,而B组呈典型的凋亡形态学改变,A组中未见凋亡性改变;B组肾小球基底膜增厚、系膜区域扩大,C组病变减轻;16周末,C组24 h尿白蛋白定量较B组显著下降(P=0.000)。结论川芎嗪能通过抑制醛糖还原酶活性,调控凋亡相关基因Bcl-2及Bax的表达而抑制细胞凋亡,明显改善糖尿病大鼠肾脏结构和功能,并可减少尿蛋白,延缓糖尿病肾病的发生和发展;高糖状态使醛糖还原酶过度激活促进凋亡相关基因Bcl-2和Bax的表达,诱导肾小管、肾小球细胞凋亡而参与糖尿病肾病的发生、发展。

Objective To research the therapeutical significance of ligustrazine for diabetic nephropathy. Methods 30 Wistar rats were selected,in which 10 rats were randomly taken as the normal control group（ A） and remaining 20 rats were given streptozotocin（ STZ）60 mg / kg） by intraperitoneal injection. After inducing diabetes, these model rats were equally divided into the two groups. The ligustrazine treatment group（ C） was given ligustrazine 100 mg /（ kg · d）, while the diabetes control group（ B） and the normal control group were given normovolemic normal saline 1 m L / 100 g. Blood sugar and 24 h urine albumin quantity were measured at 4,8,12,16 weeks. The rats were killed after 16 weeks for separating kidney. The aldose reductase（ AR） activity in tissue was detected. The immunohistochemical method was adopted to observe the expression of Bcl- 2 and Bax. The partial renal cortex was taken for conducting the pathological observation and the electronic microscopic observation on the morphologic change of renal cell apoptosis. Results The expression of Bcl- 2 protein in the ligustrazine treatment group was increased compared with the diabetes control group, while the expression of Bax protein was decreased; the AR activity in the ligustrazine treatment group was significantly decreased compared with the diabetes control group（ P = 0. 000）,and higher than that in the normal control group without statistical difference（ P〉0. 05）. The blood sugar and AR activity in the diabetes control group were significantly higher than those in the normal control group（ P〈0. 01）. The apoptosis morphology change under electronic microscopy in the ligustrazine treatment group was unapparent, while the diabetes control group revealed the typical apoptosis morphological change,but no apoptosis change was found in the normal control group; the glomerular basement membrane in the diabetes control group was thickened and the mesangial area was enlarged, the lesions in the ligustrazine treatment group were alleviated; 24 h urine albumin quantity at 16 weekend in the ligustrazine treatment group was significantly decreased compared with diabetes control group（ P = 0. 000）. Conclusion Ligustrazine can regulate the expression of apoptosisi related gene Bcl- 2 and Bax by inhibiting AR activity,inhibit apoptosis,obviously improve the renal structure and function in the diabetic rats,reduce the urine protein and delay the occurrence and development of diabetic nephropathy（ DN）; the high glucose status makes the AR excessive activation, promotes the expression of apoptosis related gene Bcl- 2 and Bax, induces the renal tubule and glomerular cell apoptosis and participates in the occurrence and dev elopment of DN.