摘要
目的观察黄芪甲苷(AS-IV)对肥胖糖尿病(DM)大鼠肾脏氧化应激、Nrf2蛋白表达和胰岛β细胞的影响。方法 2018年1-6月于武汉大学人民医院动物实验室进行实验。最终纳入健康雄性Wistar大鼠24只,随机数字表法分为对照组(n=8)、DM组(n=8)、AS-Ⅳ组(n=8),DM组给予链脲佐菌素(STZ)60 mg/kg一次性单剂量腹腔注射建立DM模型,AS-Ⅳ组在DM组基础上给予AS-Ⅳ80 mg·kg^(-1)·d^(-1)灌胃给药,8周后处死大鼠,检测各组.血尿相关指标、体质量及肾脏氧化应激指标,并观察胰岛细胞病理改变与胰岛细胞凋亡情况,采用Western免疫印迹检测Nrf 2蛋白。结果 (1)DM组及AS-Ⅳ组空腹血糖(FPG)、24h尿白蛋白排泄率(UAER)及体质量显著高于对照组,空腹胰岛素(FINS)显著低于对照组(F=59. 526、69. 342、19. 272、37. 934,P均=0. 000); AS-Ⅳ组FPG、UAER及体质量显著低于DM组,FINS显著高于DM组(P<0. 01)。(2) DM组及AS-Ⅳ组丙二醛(MDA)含量显著高于对照组,总超氧化物歧化酶(T-SOD)活性显著低于对照组(F=29. 581、53. 546,P <0. 01); AS-Ⅳ组MDA含量显著低于DM组,T-SOD活性显著高于DM组(P <0.01)。(3)与对照组比较,DM组胰岛细胞数目显著减少,且出现肿胀、坏死及空泡变性,呈不规则排列,AS-Ⅳ组胰岛细胞较DM组显著改善;DM组及AS-Ⅳ组大鼠胰岛细胞凋亡率显著高于对照组(t=9.991、5.987,P均=0.000),AS-Ⅳ组大鼠胰岛细胞凋亡率显著低于DM组,差异均有统计学意义(t=6.952,P=0. 000)。(4) Western免疫印迹结果显示,DM组大鼠肾组织总Nrf 2、核Nrf 2蛋白表达显著低于对照组及AS-Ⅳ组(t=2.542、3.881、2.642、2.267,P=0.024、0.002、0.019、0.040),AS-Ⅳ组大鼠肾组织总Nrf 2、核Nrf 2蛋白表达显著高于DM组,差异均有统计学意义(t=2.985、2.928,P=0.010、0.011)。结论 AS-Ⅳ可有效减轻肥胖DM大鼠肾脏氧化应激反应,上调Nrf2蛋白表达,并延缓胰岛β细胞凋亡,其作用机制可能与Nrf2-ARE通路被激活相关。
Objective To observe the effects of astragaloside IV (AS-IV) on oxidative stress, Nrf 2 protein expression and islet beta cells in obese diabetic rats. Methods The experiment was carried out in the Animal Laboratory of People's Hospital of Wuhan University from January to June 2018. Twenty-four healthy male Wistar rats were randomly divided into control group ( n =8), DM group ( n =8) and AS-IV group ( n =8). In the DM group, 60 mg/kg streptozotocin (STZ) was administered intraperitoneally at a single dose to establish the DM model. The AS IV group was given 80 mg·kg -1 ·d -1 of AS-IV on the basis of DM group, and then intragastrically administered. The rats were killed 8 weeks later. The indexes of blood and urine, body mass and oxidative stress of kidney were measured. The pathological changes of islet cells and apoptosis of islet cells were observed. Western blotting was used to detect Nrf 2 protein. Results (1) The fasting blood glucose (FPG), 24-hour urinary albumin excretion rate (UAER) and body mass of DM and AS-IV groups were significantly higher than those of control group, fasting insulin (FINS) were significantly lower than those of control group ( F =59.526, F =69.342, F =19.272, F =37.934, P =0.000); the FPG, UAER and body mass of AS-IV subgroup were significantly lower than those of DM group, FINS was significantly lower than those of AS-IV group. It was higher than that in DM group ( P <0.01).(2) Malondialdehyde (MDA) content in DM group and AS-IV group was significantly higher than that in control group, total superoxide dismutase (T SOD) activity was significantly lower than that in control group ( F =29.581, F =53.546, P < 0.01 ); MDA content in AS-IV group was significantly lower than that in DM group. The activity of T SOD was significantly higher than that of DM group ( P <0.01).(3) Compared with the control group, the number of islet cells in DM group decreased significantly, and there were swelling, necrosis and vacuolar degeneration, which showed irregular arrangement. The apoptotic rate of islet cells in AS-IV group was significantly higher than that in DM group and AS-IV group ( t =9.991, t = 5.987 , P =0.000). The apoptotic rate of islet cells in AS-IV group was significantly lower than that in DM group ( t =6.952, P =0.000).(4) Western blotting showed that the expression of total Nrf 2 and nuclear Nrf 2 in kidney tissue of rats in DM and AS-IV groups was significantly lower than that in control group ( t =2.542, t =3.881, t =2.642, t =2.267, P =0.024, P =0.002, P =0.019, P =0.040). The expression of total Nrf 2 and nuclear Nrf 2 in AS-IV group was significantly higher than that in DM group ( t =2.985, t =2.928, P =0.010, P =0.011). Conclusion AS IV can effectively alleviate oxidative stress in the kidney of obese DM rats, up-regulate the expression of Nrf2 protein, and delay the apoptosis of islet beta cells. Its mechanism may be related to the activation of Nrf2 ARE pathway.
作者
刘文
章尹岗
别志霞
解为慈
叶迎春
LIU Wen;ZHANG Yingang;BIE Zhixia;XIE Weici;YE Yingchun(Department of Endocrinology, Tianmen First People's Hospital, Tianmen 431700,China)
出处
《疑难病杂志》
CAS
2019年第1期67-71,共5页
Chinese Journal of Difficult and Complicated Cases
作者简介
通信作者:章尹岗,E-mail:haloyadabe@126.com
