摘要
目的观察葡萄籽原花青素提取物预灌胃对造影剂诱导糖尿病大鼠急性肾损伤的预防作用,并探讨可能作用机制。方法50只SD肥胖大鼠,腹腔注射1%链脲佐菌素(40 mg/kg),41只成功建成糖尿病大鼠模型,随机分为DM组8只、CM组9只、葡萄籽原花青素提取物低剂量组8只、中剂量组8只、高剂量组8只,另取10只肥胖大鼠为空白对照组(NC组),1 mL/kg腹腔注射柠檬酸缓冲液;低、中、高剂量组大鼠每日分别用50、250、500 mg/kg的葡萄籽原花青素提取物灌胃1次,连续3天,第3天灌胃24 h时尾静脉注射碘海醇(1.8 g I/kg);NC组、DM组、CM组大鼠每日用10 mL/kg生理盐水灌胃1次,第3天灌胃24 h时,NC组、DM组尾静脉注射5 ml/kg生理盐水;CM组尾静脉注射碘海醇(1.8 g I/kg)。末次给药48 h时各组大鼠断尾采血,检测血清肌酐(SCr)和尿素氮(BUN),采血后处死各组大鼠,取肾组织检测肾组织氧化应激指标超氧化物歧化酶(SOD)、丙二醛(MDA),采用原位缺口末端标记法测算各组大鼠肾小管上皮细胞凋亡指数,采用Western Blotting法检测各组大鼠肾组织核因子E2相关因子2(Nrf2)-Kelch样ECH关联蛋白1(Keap1)通路相关醌氧化还原酶1(NQO1)、血红素单加氧酶-1(HO-1)、Nrf2、Keap1蛋白。结果与NC组比较,CM组及低剂量组血清SCr、BUN水平高(P均<0.05)。与CM组比较,NC组、DM组、低中高剂量组血清SCr、BUN水平低(P均<0.05);与低剂量组比较,中、高剂量组大鼠血清SCr、BUN水平低(P均<0.05)。与NC组比较,DM组、CM组、低中剂量组肾组织匀浆SOD水平低,DM组、CM组及低剂量组肾组织匀浆MDA水平高(P均<0.05)。与CM组比较,DM组、低中高剂量组肾组织匀浆组SOD水平高、MDA水平低(P均<0.05)。与CM组比较,中、高剂量组大鼠肾小管上皮细胞凋亡指数小(P均<0.05)。与NC组比较,CM组大鼠肾组织Nrf2、HO-1及NQO1蛋白相对表达量低,中高剂量组大鼠肾组织Nrf2、HO-1及NQO1蛋白相对表达量高(P均<0.05);与CM组比较,低中高剂量组大鼠肾组织Nrf2、HO-1及NQO1蛋白相对表达量高(P均<0.05);与中剂量组比较,低剂量组大鼠肾组织Nrf2、HO-1及NQO1蛋白相对表达量低(P均<0.05)。结论葡萄籽原花青素提取物可改善尾静脉注射造影剂糖尿病大鼠的肾损伤程度,且500 mg/kg时效果最好。葡萄籽原花青素提取物可能通过激活Nrf2—Keap1信号通路,预防造影剂诱导的糖尿病大鼠的急性肾损伤。
Objective To observe the preventive effect of intragastric pre-administration of grape seed proanthocyan⁃idin extract(GSPE)on rats with acute renal injury induced by contrast media and to explore its possible mechanism.Methods Fifty SD obese rats were intraperitoneally injected with 1%STZ(40 mg/kg),of which,41 diabetes model rats were successfully established.They were randomly divided into five groups;there were 8 rats in the diabetes mellitus(DM)group,9 rats in the contrast medium(CM)group,8 rats in the low-dose GSPE group,8 rats in the medium-dose GSPE group,and 8 rats in the high-dose GSPE group.Another 10 obese rats were taken as the blank control(NC)group,and 1 mL/kg of citric acid buffer was intraperitoneally injected.The rats in the low-,medium-and high-dose GSPE groups were given 50,250 and 500 mg/kg GSPE by gavage once a day for three consecutive days,and the rats on the third day were given iohexol(1.8 g I/kg)by tail vein at the time of 24 hours of gavage.The rats in NC group,DM group and CM group were given 10 mL/kg normal saline once a day.When the rats were given 10 mL/kg normal saline for 24 h on the third day,the rats in the NC group and DM group were given 5 mL/kg normal saline through the tail vein,and the rats in the CM group were given iohexol(1.8 g I/kg)through the tail vein.At 48 h after the last administration,blood was taken from their tails of rats in each group,and serum creatinine(SCr)and blood urea nitrogen(BUN)were measured.Rats in each group were killed after the blood was taken.The renal tissue was taken to detect the oxidative stress indicators of renal tissues,such as superoxide dismutase(SOD)and malondialdehyde(MDA).The apoptosis index of renal tubular epitheli⁃al cells in each group was measured by in situ nick end labeling(TUNEL)method;Western blotting was used to detect the nuclear factor E2-associated factor 2(Nrf2)-Kelch like ECH-associated protein 1(Keap1)pathway-related quinone oxido⁃reductase 1(NQO1),heme monooxygenase-1(HO-1),Nrf2,and Keap1 proteins in the renal tissues of rats in each group.Results Compared with the NC group,the levels of SCr and BUN in the CM group and low-dose GSPE group were higher(all P<0.05).Compared with the CM group,the levels of SCr and BUN in the NC group,DM group and low-,medium-and high-dose GSPE groups were lower(all P<0.05).Compared with the low-dose GSPE group,the levels of SCr and BUN in the medium-dose and high-dose GSPE groups were lower(all P<0.05).Compared with the NC group,the levels of SOD in the renal tissue homogenate of the DM group,CM group,low-dose and medium-dose GSPE groups were lower,and the levels of MDA in the renal tissue homogenate of the DM group,CM group and low-dose GSPE group were higher(all P<0.05).Compared with the CM group,the levels of SOD in the DM group,low-dose,medium-dose and high-dose GSPE groups were higher,and the levels of MDA were lower(all P<0.05).Compared with the CM group,the apoptosis indices of renal tubular epithelial cells in the medium-dose and high-dose GSPE groups were lower(all P<0.05).Compared with NC group,the relative expression levels of Nrf2,HO-1,and NQO1 proteins in the renal tissues of rats in the CM group were lower,while the relative expression of Nrf2,HO-1 and NQO1 proteins in the renal tissues of rats in the medium-dose and high-dose GSPE group were higher(all P<0.05).Compared with the CM group,the relative ex⁃pression levels of Nrf2,HO-1,and NQO1 proteins in the kidney tissues of rats in the low-dose,medium-dose and high-dose GSPE group were higher(all P<0.05).Compared with the medium-dose group,the relative expression levels of Nrf2,HO-1,and NQO1 proteins in the kidney tissues of rats in the low-dose group were lower(all P<0.05).Conclu⁃sion GSPE can alleviate renal injury in diabetes rats injected with contrast agent through tail vein,and the optimum con⁃centration is 500 mg/kg.GSPE may prevent acute renal injury in contrast-induced diabetes rats by activating Nrf2-Keap1 signaling pathway.
作者
翟志红
张海俊
黄辉
牛强
ZHAI Zhihong;ZHANG Haijun;HUANG Hui;NIU Qiang(Department of Cardiology,The First Affiliated Hospital of Medical College of Shihezi University,Shihezi 832000,China;不详)
出处
《山东医药》
CAS
2023年第6期19-23,共5页
Shandong Medical Journal
基金
国家自然科学基金资助项目(81860559)
石河子大学自主资助支持校级立项项目(ZZZC2022078)。
