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中国应用生理学杂志 ›› 2019, Vol. 35 ›› Issue (1): 60-64.doi: 10.12047/j.cjap.5589.2019.015

• 研究论文 • 上一篇    下一篇

口服AdipoRon对2型糖尿病小鼠脾脏和胰腺功能的影响

谢克俭,黄玲,屈小虎,李雪,王少杰,肖敏   

  1. 温州医科大学检验医学院、生命科学学院, 浙江 温州 325035
  • 收稿日期:2018-04-10 出版日期:2019-01-28 发布日期:2019-06-27
  • 通讯作者: Tel: 13600673947; E-mail: 840855242@qq.com
  • 基金资助:
    2015年度浙江省公益技术应用研究计划(实验动物)(2015C37099)

Effects of AdipoRon orally on the functions of spleen and pancreas in type 2 diabetic mice

XIE Ke-jian, HUANG Ling, QU Xiao-hu, LI Xue, WANG Shao-jie, XIAO Min   

  1. Department of Biology, School of Laboratory Medicine and Life Science, Wenzhou Medical University, Wenzhou 325035 China
  • Received:2018-04-10 Online:2019-01-28 Published:2019-06-27

摘要: 目的:观察口服AdipoRon对2型糖尿病小鼠脾脏和胰腺功能的影响,为AdipoRon的临床应用提供基础资料。方法:将40只C57/BL6雄性小鼠随机分为正常对照组(NC, n=10)和造模组(n=30),并分别给予普通饲料和高脂高糖饲料喂养。4周后,造模组小鼠腹腔注射链脲佐菌素(STZ,40 mg/kg)以诱导建立2型糖尿病模型。造模成功后将糖尿病模型小鼠随机分为糖尿病模型组(DM)、高剂量AdipoRon(50 mg/kg)(DM+H)组、低剂量AdipoRon(20 mg/kg)(DM+L)组,每组10只。DM+L组和DM+H组灌胃相应浓度的AdipoRon(使用去离子水溶解AdipoRon),NC组和DM组灌胃等体积去离子水,每日灌胃1次,灌胃10 d。末次干预后禁食12 h,处死小鼠取血液、胰腺和脾脏。HE染色光镜下观察胰腺的病理改变;ELISA法检测小鼠胰腺和脾脏组织中胰岛素受体(INSR)、胰岛素受体底物1(IRS-1)和肿瘤坏死因子-α(TNF-α)蛋白质含量;小鼠脾脏系数;Western blot法检测胰腺组织中p-IRS-1蛋白质水平;实时荧光定量PCR检测胰腺组织中insulin mRNA 表达。结果:光镜下可见正常组小鼠胰腺组织排列紧密、饱满、胰岛体积大,DM组小鼠胰腺组织排列较为疏散、胰岛体积较小,口服AdipoRon组小鼠胰腺组织基本紧密、饱满、胰岛体积略小。与NC比较,DM组小鼠胰腺和脾脏TNF-α水平明显升高,INSR、IRS-1水平均降低,脾脏系数、胰腺p-IRS-1蛋白质水平和insulin mRNA表达均降低,均具有统计学意义(P<0.05);与DM组比较,口服AdipoRon组小鼠胰腺和脾脏TNF-α水平明显下降,INSR和IRS-1水平均升高,脾脏系数升高,DM+H组胰腺p-IRS-1蛋白质水平和insulin mRNA 表达均升高(P<0.05);与DM+L组比较,DM+H组小鼠TNF-α水平明显下降,INSR和IRS-1水平均升高(P<0.05)。结论:口服AdipoRon可通过减弱糖尿病小鼠炎症反应,上调INSR表达、提高p-IRS-1水平,从而对糖尿病小鼠脾脏和胰腺组织有一定的保护作用。

关键词: AdipoRon, 2型糖尿病, 胰腺, 脾脏, 小鼠

Abstract: Objective:To observe the effects of AdipoRon orally on the functions of spleen and pancreas in type 2 diabetic mice, in order to present data for clinical application.Methods: Forty C57/BL6 male mice were randomly divided into 2 groups: normal control group (n=10) and model group (n=30), the former group was fed normally, while the later group was fed with high fat and sugar for 4 weeks.After that, type 2 diabetes model was established in DM group induced by intraperitoneal injection of streptozotocin (STZ, 40 mg/kg).As type 2 diabetes model established successfully, the model mice were randomly divided into three groups (n=10): diabetes mellitus (DM) group, high dose of AdipoRon group (DM + H) and low dose of adiponRon group (DM + L).All the four groups were treated with saline, saline, AdipoRon at the doses of 20 mg/kg and 50 mg/kg by gavages respectively, once a day for 10 days.And then put them to death for collecting blood, pancreas and spleen.Pathological changes of pancreas were observed with a light microscope after HE staining.Protein contents of insulin receptor (INSR), insulin receptor substrate 1( IRS-1) and tumor necrosis factor-α(TNF-α) in pancreatic and spleen tissues were detected by ELISA.The protein level of phosphorylation insulin receptor substrate 1(p-IRS-1) in pancreas was determined by Western blot, and the expression of insulin mRNA in pancreas was tested by RT-PCR.Results: Under the light microscope, it was visible that the pancreatic tissue in NC group was full and closely packed, and the islet was big.Pancreatic tissue of DM mice was incompact and the islet of DM mice was smaller than that of normal mice.As for the mice treated with AdipoRon orally, the pancreatic tissue was full and closely arranged, and the islet was slightly smaller.Compared with NC group, the levels of TNF-α in pancreas and spleen of DM group were increased markedly, the levels of INSR and IRS-1 were decreased, the spleen coefficient, p-IR-1 protein level and insulin mRNA expression in pancreas were decreased, all were significant statistically (P<0.05).Compared with DM group, the levels of TNF-α in pancreas and spleen of AdipoRon groups were decreased, the levels of INSR and IRS-1 in pancreas and spleen of AdipoRon groups were increased, while the spleen coefficient was increased (P<0.05).The p-IRS-1 protein level and insulin mRNA expression in pancreas in DM+H group were increased (P<0.05).Compared with DM + L group, the level of TNF-α was decreased, and the levels of INSR and IRS-1 were significantly increased (P<0.05) in DM + H group (P<0.05).Conclusion: Oral administration of AdipoRon can protect the spleen and pancreas of diabetic mice by decreasing the inflammatory response, up-regulating the expression of INSR, and increasing p-IRS-1 level in diabetic mice.

Key words: AdipoRon, type 2 diabetes, pancreas, spleen, mouse

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