摘要
目的探究小豆蔻素(CAR)调节核因子e2相关因子2/血红素加氧酶-1/核因子-κB(Nrf2/HO-1/NF-κB)信号通路对妊娠糖尿病(GDM)大鼠氧化应激损伤的影响。方法实验分为对照组(Control组)、妊娠糖尿病模型组(GDM组)、低剂量CAR组(L-CAR组)、高剂量CAR组(H-CAR组),高剂量CAR+Nrf2抑制剂Brusatol组(H-CAR+BR组),每组12只。采用血糖仪测定大鼠空腹血糖(FBG);胰岛素放射免疫分析试剂盒检测大鼠胰岛素(FINS)水平;试剂盒检测血清天门冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)、碱性磷酸酶(ALP)以及总抗氧化能力(T-AOC);试剂盒测定肝脏组织谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)、丙二醛(MDA)、过氧化氢酶(CAT)水平;ELISA法测定血清炎性因子-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)、白细胞介素-10(IL-6)水平;H-E染色法检测肝脏组织病理变化;western blot法检测大鼠肝脏组织Nrf2、HO-1、NF-κB p65、p-NF-κB p65蛋白水平。结果与Control组比较,GDM组大鼠肝小叶及肝窦组织结构紊乱,且大量细胞变性坏死,FBG水平(1 d、7 d、14 d)、AST、ALT、ALP、MDA、TNF-α、IL-6、IL-1β、p-NF-κB p65/NF-κB p65升高,FINS水平(1d、7 d、14 d)、T-AOC、GSH-Px、CAT、SOD、IL-10、Nrf2、HO-1水平降低(P<0.05);与GDM组比较,L-CAR组、H-CAR组大鼠肝小叶及肝窦组织结构好转且坏死细胞减少,FBG水平(7d、14d)、氧化应激水平、促炎因子含量及p-NF-κBp65/NF-κBp65降低,FINS水平(7d、14d)、Nrf2、HO-1水平升高(P<0.05),且H-CAR组上述指标变化较L-CAR组显著(P<0.05);Nrf2抑制剂Brusatol可削弱高剂量CAR对GDM大鼠氧化应激损伤的缓解作用(P<0.05)。结论CAR可能通过上调Nrf2/HO-1表达,抑制NF-κB通路,进而减轻GDM大鼠氧化应激损伤。
Objective To explore the influence of cardamonin(CAR)on oxidative stress injury in gestational diabetes mellitus(GDM)rats by regulating nuclear factor e2-related factor 2/heme oxygenase-1/nuclear factor-κB(Nrf2/HO-1/NF-κB)signaling pathway.Methods The experiment was separated into Control group,gestational diabetes mellitus model group(GDM group),low-dose CAR group(L-CAR group),high-dose CAR group(H-CAR group),high-dose CAR+Nrf2 inhibitor Brusatol group(H-CAR+BR group),12 animals in each group.The fasting blood glucose(FBG)of rats was measured by a blood glucose meter,insulin radioimmunoassay kit was performed to detect the level of insulin in rats(FINS).The kit was applied to detect serum aspartate aminotransferase(AST),alanine aminotransferase(ALT),alkaline phosphatase(ALP)and total antioxidant capacity(T-AOC).The kit was applied to measure the levels of glutathione peroxidase(GSH-Px),superoxide dismutase(SOD),malondialdehyde(MDA)and catalase(CAT)in liver tissue.The levels of serum inflammatory factors-tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6),and interleukin-10(IL-6)was measured by ELISA.H-E staining was applied to detect the pathological changes of liver tissue.Western blot was applied to detect the protein levels of Nrf2,HO-1,NF-κB p65 and p-NF-κB p65 in rat liver tissue.Results Compared with the Control group,the hepatic lobules and hepatic sinusoids of the GDM group were disordered,and a large number of cells were degenerated and necrotic,the levels of FBG(1 d,7 d,14 d),AST,ALT,ALP,MDA,TNF-α,IL-6,IL-1β,p-NF-κB p65/NF-κB p65 increased,the levels of FINS(1 d,7 d,14 d),T-AOC,GSH-Px,CAT,SOD,IL-10,Nrf2,and HO-1 decreased(P<0.05).compared with the GDM group,the tissue structure of the hepatic lobules and hepatic sinusoids of the L-CAR group and the H-CAR group improved and the necrotic cells decreased,the level of FBG(7 d,14 d),oxidative stress level,content of pro-inflammatory factors and p-NF-κB p65/NF-κB p65 decreased,the levels of FINS(7 d,14 d),Nrf2 and HO-1 increased(P<0.05),and the changes of the above indicators in the H-CAR group were obviously higher than those in the L-CAR group(P<0.05).The Nrf2 inhibitor Brusatol attenuated the alleviation effect of high-dose CAR on oxidative stress injury in GDM rats(P<0.05).Conclusion CAR may reduce oxidative stress injury in GDM rats by up-regulating the expression of Nrf2/HO-1 and inhibiting the NF-κB pathway.
作者
孙芙林
岳涛
刘海燕
郭菊芳
SUN Fulin;YUE Tao;LIU Haiyan;GUO Jufang(Maternity Health Department,Jinniu Maternity and Child Health Hospital of Chengdu,Chengdu,Sichuan 610036,China;Department of Laboratory Medicine,the Pengzhou Hospital of Integrated Traditional Chinese and Western Medicine,Chengdu,Sichuan 611930,China)
出处
《中国优生与遗传杂志》
2023年第2期269-274,共6页
Chinese Journal of Birth Health & Heredity
基金
中国疾病预防控制中心妇幼保健中心2020年度“母婴营养与健康研究项目”(2020FYH019)。
