白藜芦醇对妊娠期糖尿病大鼠肾脏组织抗氧化和抗炎的作用

Anti-oxidation and Anti-inflammatory Effects of Resveratrol on Renal Tissue in Gestational Diabetic Rats

目的:本实验以妊娠期糖尿病大鼠为研究对象,探讨白藜芦醇对妊娠期糖尿病大鼠肾脏组织抗氧化和抗炎的作用。方法:实验用雌性大鼠经高脂高糖饲料喂养8周后与雄鼠合笼过夜制备孕鼠,5 d后采用ip链脲佐菌素(STZ,35 mg·kg^(-1))破坏胰岛β细胞的方法诱导制备糖尿病大鼠,选取80只模型大鼠根据血糖水平随机分为模型组、白藜芦醇60,120,240 mg·kg^(-1)组治疗组和盐酸二甲双胍(200 mg·kg^(-1))阳性药组,并另取16只同批妊娠5 d大鼠作为正常妊娠组,以及16只同龄非妊娠雌性大鼠作为正常非妊娠组;判定造模成功后立即ig给药,每天1次,疗程2周。给药治疗前和治疗完成后分别观察各组大鼠一般状况;治疗完成后测定各组大鼠空腹血糖水平,测量24 h尿量并测定24 h尿蛋白量(UPro),检测血清中血尿素氮(BUN),肌酐(SCr),尿酸(UA)含量,通过PAS染色观察肾脏组织病变,通过原位末端转移酶标记法(TUNEL)染色观察肾脏组织细胞凋亡状况并计算凋亡指数;测定血清中总抗氧化能力(T-AOC)水平和肾脏组织中超氧化物歧化酶(SOD),谷胱甘肽过氧化物酶(GSH-Px),过氧化氢酶(CAT)活性以及丙二醛(MDA)含量,检测血浆中炎症因子C-反应蛋白(CRP),肿瘤坏死因子-α(TNF-α),白细胞介素-1β(IL-1β),白细胞介素-6(IL-6),细胞间黏附分子-1(ICAM-1)水平。结果:正常妊娠组大鼠各监测指标与正常非妊娠组比较均无显著性差异;与正常妊娠组比较,模型组大鼠"三多一少"症状明显,血糖水平显著升高,24 h尿量显著增多,UPro显著增高,血清中BUN,SCr,UA含量显著升高,呈现明显的肾脏组织病变和肾细胞凋亡、凋亡指数显著升高,血清中T-AOC水平和肾脏组织中SOD,GSH-Px,CAT活性均显著降低,MDA含量显著升高,上述差异均具有统计学意义(P<0.05,P<0.01);与模型组比较,白藜芦醇治疗组大鼠一般状况明显改善,"三多一少"症状明显减轻;白藜芦醇120,240 mg·kg-1治疗组大鼠空腹血糖水平显著降低,24 h尿量显著减少,UPro显著降低,血清中BUN,SCr,UA含量显著降低,肾脏组织病变和肾细胞凋亡状况均明显改善、凋亡指数显著降低,肾脏组织中SOD,CAT活性显著升高且MDA含量显著降低,并且240 mg·kg-1治疗组大鼠血清中T-AOC水平和肾脏组织中GSH-Px活性均显著升高,上述差异均具有统计学意义(P<0.05,P<0.01)。结论:白藜芦醇对妊娠期糖尿病大鼠肾脏组织抗氧化和抗炎能力具有改善作用。

Objective： To investigate the anti-oxidation and anti-inflammatory effects of resveratrol（ Res） on renal tissues in gestational diabetic rats. Method： Female rats were fed with high fat and high sugar diet for 8 weeks and then kept in the same cage with male rats to prepare pregnant rats. After 5 days,the pregnant rats were injected with STZ（ 35 mg·kg^-1） by intraperitonal to make gestational diabetic models. 80 model rats were selected andrandomly devided into model group, Res（ 60, 120, 240 mg ·kg^-1） groups and Metformin hydrochloride（ 200 mg·kg^-1） group. Another 16 gestational rats in the same batch were selected as normal gestational group,and 16 non-gestational female rats at the same age were selected as normal non-gestational group. After successful modeling,the medicines were given by intragastric administration for 2 weeks,once a day.Before and after treatment,the general status of rats in every group were observed. After treatment,the levels of blood sugar were determined; the level of 24 h urine protein（ UPro） was determined; the levels of blood urea nitrogen（ BUN）, creatinine（ SCr）, and uric acid（ UA） in serum were determined; the renal tissue histopathological changes were observed by periodic acid-schiff staining（ PAS） method; the renal cells apoptosis was observed by terminal-deoxynucleoitidyl transferase mediated nick end labeling（ TUNEL） staining method,and the apoptosis index was alculated; the levels of total antioxidative capacity（ T-AOC） in serum and the activities of superoxide dismutase（ SOD）,glutathione peroxidase（ GSH-Px）,catalase（ CAT） as well as the content of malondialdehyde（ MDA） in renal tissues were determined; the levels of C-reactionprotein（ CRP）,tumor nerosis factor-α（ TNF-α）,interlukin-1β（ IL-1β）,interlukin-6（ IL-6）,and intercellular cell adhesion molecule-1（ ICAM-1） in plasma were also determined. Result： There were no differences in various monitoring indexes between the normal gestation group and normal non-gestation group. As compared with the normal gestation group,the blood sugar level was significantly increased in model group（ P〈0. 01）; the 24 h urine volume,UPro and the contents of BUN,SCr,UA in serum were significantly increased（ P〈0. 01）; with obvious histopathological changes in renal tissues and renal cells apoptosis,and the apoptosis index was significantly increased（ P〈0. 01）;the level of T-AOC and the activities of SOD,GSH-Px,CAT in renal tissues were significantly decreased,and the content of MDA in renal tissue was significantly increased（ P〈0. 01）. As compared with the model group,the general status of the rats in Res groups were improved; the symptoms were relieved; the levels of blood sugar in Res120 and 240 mg·kg^-1groups were significantly decreased; the 24 h urine volume and the UPro level were significantly decreased, and the contents of BUN, SCr, UA in serum were significantly decreased; the histopathological changes in renal tissues and renal cells apoptosis were significantly improved,and the apoptosis indexes were significantly decreased; the activities of SOD,CAT in renal tissues were significantly increased,and the content of MDA was significantly decreased; the levels of T-AOC and the activity of GSH-Px in renal tissues of Res 240 mg·kg^-1group were significantly increased（ P〈0. 05,P〈0. 01）. Conclusion： Res had anti-oxidation and anti-inflammatory effects on renal tissues in diabetic rats.