山楂叶总黄酮对2型糖尿病大鼠肾脏组织保护作用的研究

Protective Effect of Hawthorn Leaves Favonoids on the Kidneys of Type 2 Diabetic Rats

目的:研究山楂叶总黄酮(Hawthorn Leaves Favonoids,HLF)对糖尿病并发肾脏组织损伤的抑制作用。方法:96只实验用大鼠按随机数字表法分为正常对照组、模型组、HLF低(50 mg/kg)、中(100mg/kg)、高(200 mg/kg)剂量组和阳性对照组(盐酸二甲双胍,200 mg/kg),每组16只。除正常对照组外,其余各组大鼠均采用链脲佐菌素破坏胰岛β细胞的方法复制实验用2型糖尿病大鼠模型;确认造模成功后,各组大鼠分别灌胃给药治疗,每天1次,疗程6周。治疗完成后,测定各组大鼠空腹血糖水平,称量体质量并计算肾脏指数;测量24 h尿量及24 h尿蛋白量(UPro),测定血清中血尿素氮(BUN)、肌酐(SCr)、尿酸(UA)水平,肾脏组织中总抗氧化能力(T-AOC)、抗氧化酶活性以及丙二醛(MDA)含量;通过HE染色观察肾脏组织机构形态学变化;末端标记法(TUNEL)检测肾脏组织细胞凋亡状况并计算凋亡指数(Apoptosis Index,AI);采用酶联免疫法(ELISA)测定血浆中肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)、白介素-6(IL-6)和细胞间黏附分子-1(ICAM-1)含量水平。结果:经HLF治疗6周后,山楂叶总黄酮能够显著降低实验用2型糖尿病大鼠空腹血糖水平、改善体质量并显著降低肾脏指数,显著减少24 h尿量、降低UPro,显著降低血清中BUN、SCr、UA含量,显著提高肾组织中TAOC水平并显著改善超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性、显著降低MDA含量,上述差异均具有统计学意义(P<0.05,P<0.01);HLF能够剂量依赖性地改善2型糖尿病大鼠肾脏组织病理性形态学改变及细胞凋亡状况,HLF中、高剂量组AI较2型糖尿病模型对照组显著降低(P<0.01);HLF能够显著降低2型糖尿病大鼠血浆中TNF-α、IL-6、ICAM-1、IL-1β水平(P<0.05,P<0.01)。结论:HLF能够剂量依赖性地改善肾功能,抑制氧化应激损伤和细胞凋亡,降低炎性介质水平,降低细胞间黏附分子,改善肾脏组织病变,提示HLF对2型糖尿病大鼠肾脏组织具有剂量依赖性的保护作用。

Objective： To investigate the protective effect of hawthorn leaves favonoids on the kidneys of type 2 diabetic rats. Methods： Ninety- six experimental rats were randomly devided into 6 groups： normal control group,model group,HLF low（50 mg / kg）,medium（100 mg / kg）,high（200 mg / kg） dose groups and metformin hydrochloride 200 mg / kg treatment group; except the rats in normal control group,others were injected STZ to make type 2 diabetic model rats; the drugs were given by intragastric administration for 6 weeks,once a day. Six weeks later,the level of blood sugar was determined; the body weight was determined and the renal index（RI） was calculated; the urine volume and the UPro were determined; the contents of BUN,SCr,UA in serum were determined; the activity of SOD,GSH- Px,T- AOC and the content of MDA in renal tissue were determined; the histopathological changes of renal tissue were observed by HE staining and the apoptosis of renal tissue cells was observed by TUNEL; the levels of TNF- α,IL- 1β,IL- 6,ICAM- 1 in plasma were determined. Results： Compared with the type 2 diabetic models in the control group,the blood sugar levels of HLF medium and high dose groups were significantly decreased; the body weight was significantly increased and the RI was significantly decreased; the urine volume and the UPro were significantly decreased,and the contents of BUN,SCr and UA in serum were significantly decreased; the activities of SOD were significantly increased and the contents of MDA were significantly decreased; the activities of GSH- Px and T-AOC in HLF 200 mg / kg treatment group were significantly increased,and all of the above differences were significant（P〈0. 05,P〈0. 01）; the histopathological changes and apoptosis were significantly improved,especially the HLF 200 mg / kg treatment group,and the AI of HLF medium and high dose groups was significantly decreased（P〈0. 01）; the levels of TNF- α,IL- 6 and ICAM- 1 in plasma of medium and high dose groups were significantly decreased（P〈0. 05,P〈0. 01）,and the level of IL- 1β in HLF high dose group was significantly decreased（P〈0. 01）. Conclusion： HLF has dose- dependent effects on improving renal function,reducing the damage of oxidative stress,depressing the apoptosis,lowering the level of inflammatory mediators and the level of ICAM- 1 and improving the renal tissue histopathological changes,which suggests that HLF has dose- dependently protective effects on the kidneys in type 2 diabetic rats.