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姜黄素及其类似物J7对2型糖尿病大鼠睾丸氧化应激损伤的干预作用*
引用本文:徐菲菲,缪成锋,池琛,吴谷,陈国荣. 姜黄素及其类似物J7对2型糖尿病大鼠睾丸氧化应激损伤的干预作用*[J]. 中国应用生理学杂志, 2019, 35(2): 145-149. DOI: 10.12047/j.cjap.5783.2019.032
作者姓名:徐菲菲  缪成锋  池琛  吴谷  陈国荣
作者单位:温州医科大学附属第一医院病理科, 浙江 温州 325015
基金项目:温州市重大科技专项(2018ZY019)
摘    要:目的: 研究姜黄素(CUR)及其类似物J7对糖尿病大鼠睾丸氧化应激损伤的干预作用。方法: 60只SD大鼠随机分组,其中10只作为正常(NC)组,余50只通过高脂饮食和腹腔注射链脲佐菌素诱导建立糖尿病大鼠模型,造模成功后将其再分为4组:糖尿病(DM,n=12)组、姜黄素治疗(CUR,n=10)组、J7高剂量治疗(J+,n=10)组、J7低剂量治疗(J-,n=10)组。CUR组大鼠每天予以姜黄素20 mg/kg灌胃治疗,J+及J-组分别予以J7 20 mg/kg、10 mg/kg灌胃治疗,8周后处死大鼠,测量各组大鼠体重、空腹血糖,羟胺法和硫代巴比妥酸法分别检测超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量,Western blot法检测tNrf2、pNrf2、CAT、NQO1蛋白表达水平,qRT-PCR检测CAT、NQO1、HO1 mRNA表达水平,光镜下观察大鼠睾丸形态学改变,免疫组化检测Nrf2及CAT蛋白表达情况。结果: DM组血糖、MDA水平升高(P<0.05),体重、SOD活性、pNrf2/tNrf2、CAT、NQO1蛋白及CAT、NQO1、HO1 mRNA水平均有下降(P<0.05);光镜下见睾丸各级生精细胞减少、排列紊乱;免疫组化显示Nrf2蛋白核周表达量下降,CAT蛋白表达水平降低。经姜黄素及J7治疗后,三个治疗组的MDA水平均下降(P<0.05),SOD活性、pNrf2/tNrf2、CAT、NQO1蛋白及NQO1、HO1 mRNA水平均上升(P<0.05),J+及J-组血糖显著下降(P<0.05),J+组CAT mRNA水平显著上升(P<0.05);J+组pNrf2/tNrf2比值明显高于CUR组及J-组(P<0.05),J+组CAT蛋白水平也明显高于J-组(P<0.05),其余指标在三个治疗组间不具有显著性差异。光镜下见三个治疗组睾丸形态学病变减轻;免疫组化显示Nrf2蛋白核周表达量上升,CAT蛋白表达水平升高。提示高剂量J7抗糖尿病大鼠睾丸的氧化应激损伤的能力较强。结论: 姜黄素及J7可在一定程度上对抗糖尿病大鼠睾丸的氧化应激损伤,其机制可能与Nrf2-ARE信号通路的激活相关。

关 键 词:姜黄素  姜黄素类似物J7  糖尿病  大鼠  睾丸  氧化应激  

Intervention of curcumin and its analogue J7 on oxidative stress injury in testis of type 2 diabetic rats
XU Fei-fei,MIAO Cheng-feng,CHI Chen,WU Gu,CHEN Guo-rong. Intervention of curcumin and its analogue J7 on oxidative stress injury in testis of type 2 diabetic rats[J]. Chinese journal of applied physiology, 2019, 35(2): 145-149. DOI: 10.12047/j.cjap.5783.2019.032
Authors:XU Fei-fei  MIAO Cheng-feng  CHI Chen  WU Gu  CHEN Guo-rong
Affiliation:Department of Pathology, the First Affiliated Hospital, Wenzhou Medial University, Wenzhou 325015, China
Abstract:Objective: To investigate the intervention of curcumin and its analogue J7 on oxidative stress injury in testis of type 2 diabetic rats. Methods: Sixty male SD rats, 10 rats were chosen as normal control group (NC), the other 50 rats were assigned to experiment group. Experiment diabetic rats were induced by high-fat food and intraperitoneal injection of steptozotocin (STZ). After the model was established successfully, diabetic rats were divided into four groups randomly: diabetes mellitus group (DM, n=12), curcumin treatment group (CUR, n=10), high dose treatment group of J7 (J+, n=10), low dose treatment group of J7 (J-, n=10). The CUR group were intragastrically administered with curcumin 20 mg/kg daily, in addition, the J+ group and the J- group were intragastrically administered with J7 20 mg/kg and 10 mg/kg daily respectively. After 8 weeks, the fast blood glucose was detected biochemically. The activity of superoxide dismutase (SOD) and the level of malondialdehyde (MDA) were detected by hydroxylamine method and thiobarbituric acid method respectively. The protein expressions of the nuclear factor-erythroid 2-related factor 2 (tNrf2), phosphorylation of Nrf2 (pNrf2), catalase (CAT), NAD(P)H quinine oxidoreductase 1 (NQO1) were measured by Western blot. The mRNA expressions of CAT, NQO1, hemeoxygenase-1 (HO1) were measured by quantitative real-time PCR (qRT-PCR). Morphological structure of testis was observed by hematoxylin-eosin (HE) staining. The expressions of Nrf2 and CAT were also detected by immunohistochemical method. Results: The levels of fast blood glucose and MDA in DM group were increased significantly(P<0.05), while the body weight, the activity of SOD, the protein expressions of pNrf2/tNrf2, CAT, NQO1 and the mRNA expressions of CAT, NQO1, HO1 were decreased (P<0.05). Under light microscope, the DM group showed disrupted histological appearance. Immunohistochemistry showed that the protein expressions of Nrf2 around the nucleus and CAT were decreased. With the treatment of curcumin and J7, the MDA levels in the three treatment groups were decreased (P<0.05). The activity of SOD, the protein expressions of pNrf2/tNrf2, CAT, NQO1 and the mRNA expressions of NQO1, HO1 were increased (P<0.05). the levels of fast blood glucose were decreased in the J+ and J- group (P<0.05), and the mRNA expression of CAT was increased in the J+ group (P<0.05). The ratio of pNrf2/tNrf2 in the J+ group was significantly higher than that in CUR and J- group (P<0.05). The protein level of CAT in the J+ group was also significantly higher than that in J- group (P<0.05). There were no significant differences in other indexes among the three treatment groups. Under light microscope, the morphology was obviously improved in the three treatment groups. Immunohistochemistry showed that the protein expressions of Nrf2 around the nucleus and CAT were increased in the three treatment groups. It was suggested that high dose J7 had better antioxidant stress ability in testis of diabetic rats. Conclusion: Curcumin and J7 could inhibit the oxidative stress damage of testicular tissue in diabetic rats, which might be related with the activation of the Nrf2-ARE signaling pathway.
Keywords:curcumin   curcumin analogue J7   diabetes   rats   testis   oxidative stress  
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