橙皮素通过Nrf2-ARE通路对大鼠肾缺血再灌注损伤的保护作用  

Protective effect of hesperetin on renal ischemia-reperfusion injury in rats through Nrf2-ARE pathway

作　　者：翟鑫铎 金敏 刘宇[1] ZHAI Xin-duo;JIN Min;LIU Yu(Department of Pharmacology,Shanxi Medical University,Taiyuan 030001,Shanxi Province,China)

机构地区：[1]山西医科大学药理学教研室,山西太原030001

出　　处：《中国临床药理学杂志》2024年第12期1803-1807,共5页The Chinese Journal of Clinical Pharmacology

基　　金：山西省留学人员科技活动择优基金资助项目(20210043)。

摘　　要：目的研究橙皮素(HSP)对大鼠肾缺血再灌注损伤(RIRI)的作用,并探讨其作用机制。方法将50只雄性SD大鼠随机分为假手术组、模型组、HSP预处理组、全反式维甲酸(ATRA)预处理组(ATRA+I/R)、HSP+ATRA预处理组(HSP+ATRA+I/R),每组10只。假手术组仅打开腹腔暴露双肾。其余各组采用夹闭双侧肾蒂45 min,再用灌注24 h的方法建立RIRI模型。用苏木精-伊红(HE)染色法确定肾组织损伤水平;用微板法检测血清肌酸酐(Scr)水平;用脲酶法检测血清尿素氮(BUN)水平;用酶联免疫吸附测定(ELISA)法检测血清中白细胞介素-6(IL-6)、白细胞介素-1β(IL^(-1)β)和肿瘤坏死因子-α(TNF-α)的含量;用试剂盒法检测肾组织超氧化物歧化酶(SOD)、谷胱甘肽(GSH)、丙二醛(MDA)和过氧化氢酶(CAT)水平;用蛋白质印迹法检测肾组织中核因子E2相关因子2(Nrf2)及其下游抗氧化蛋白血红素氧合酶-1(HO-1)和醌氧化物还原酶1(NQO1)的表达。结果假手术组、模型组、HSP预处理组肾小管损伤评分分别为(0.20±0.45)、(4.20±0.84)和(2.40±0.55)分;Scr含量分别为(55.52±11.23)、(207.10±19.22)和(105.60±18.11)μmol·L^(-1);IL-6含量分别为(33.66±6.83)、(172.50±8.09)和(105.40±10.03)pg·mL^(-1)。假手术组、模型组、HSP预处理组、ATRA预处理组和HSP+ATRA预处理组SOD水平分别为(57.04±3.44)、(37.29±3.60)、(51.61±9.41)、(32.55±5.58)和(37.40±3.66)U·mg·prot^(-1);Nrf2蛋白相对表达水平分别为0.37±0.24、0.57±0.28、1.31±0.34、0.44±0.17和0.77±0.25;HO-1蛋白相对表达水平分别为0.26±0.14、0.57±0.30、1.32±0.61、0.53±0.28和0.67±0.50。模型组与假手术组相比,肾小管损伤评分、Scr、IL-6、SOD指标比较,在统计学上差异均有统计学意义(P<0.05,P<0.01);HSP预处理组与模型组相比,上述指标在统计学上差异均有统计学意义(P<0.05,P<0.01)。结论HSP可能通过激活Nrf2-抗氧化反应元件(ARE)通路上调其下游抗氧化蛋白的表达,减轻氧Objective To study the effect of hesperetin(HSP)on renal ischemia-reperfusion injury(RIRI)in rats and its mechanism.Methods Fifty male SD rats were randomly divided into five groups:sham group,model group(renal ischemia-reperfusion injury I/R),HSP pretreatment(HSP+I/R)group,all-trans retinoic acid pretreatment(ATRA+I/R)group,HSP and ATRA pretreatment(HSP+ATRA+I/R)group.Each group had 10 rats.The sham group only exposed both kidneys by opening the abdominal cavity.In other groups,the RIRI model was established by occluding the bilateral renal pedicles for 45 min,followed by reperfusion for 24 h.Hematoxylin-eosin staining(HE)was used to ascertain the extent of kidney injury.The microplate method was used to measure serum creatinine(Scr)levels,while the urease method was used to measure blood urea nitrogen(BUN)levels.Enzyme-linked immunosorbent assay(ELISA)was used to detect the contents of interleukin-6(IL-6),interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in the serum.The kit method was used to examine the levels of superoxide dismutase(SOD),glutathione(GSH),malondialdehyde(MDA)and catalase(CAT)in kidney tissue.The Western blot analysis was conducted to detect the expression of nuclear factor erythroid 2 related factor 2(Nrf2)and its downstream antioxidant proteins heme oxygenase-1(HO-1)and quinone oxide reductase 1(NQO1)in renal tissues.Results The scores for renal tubule damage in sham,I/R and HSP+I/R groups were 0.20±0.45,4.20±0.84 and 2.40±0.55;Scr contents were(55.52±11.23),(207.10±19.22)and(105.60±18.11)μmol·L^(-1);IL-6 contents were(33.66±6.83),(172.50±8.09)and(105.40±10.03)pg·mL^(-1).The SOD levels in sham,I/R,HSP+I/R,ATRA+I/R and HSP+ATRA+I/R groups were(57.04±3.44),(37.29±3.60),(51.61±9.41),(32.55±5.58)and(37.40±3.66)U·mg·prot^(-1);the relative expression levels of Nrf2 were 0.37±0.24,0.57±0.28,1.31±0.34,0.44±0.17 and 0.77±0.25;the relative expression levels of HO-1 protein were 0.26±0.14,0.57±0.30,1.32±0.61,0.53±0.28 and 0.67±0.50.There was significant difference

关 键 词：橙皮素 肾缺血再灌注损伤 炎症 氧化应激 

分 类 号：R28[医药卫生—中药学]