褪黑素对妊娠期糖尿病大鼠胰腺自噬的影响及其机制  

EFFECT OF MELATONIN ON PANCREATIC AUTOPHAGY IN RATS WITH GESTATIONAL DIABETES MELLITUS AND THE MECHANISM

作　　者：郑晴晴 李超[3] 李岩[3] 乔炳龙[4] 马春玲[3] ZHENG Qingqing;LI Chao;LI Yan;QIAO Binglong;MA Chunling(Faculty of Medicine,Qingdao University,Qingdao 266071,China)

机构地区：[1]青岛大学医学部,山东青岛266071 [2]枣庄市市中区妇幼保健院妇产科 [3]青岛大学附属医院产科 [4]青岛大学附属医院放射科

出　　处：《精准医学杂志》2023年第3期205-209,共5页Journal of Precision Medicine

基　　金：山东省医药卫生科技发展项目(2016WS0274)。

摘　　要：目的 探讨褪黑素对妊娠期糖尿病(gestational diabetes mellitus, GDM)大鼠胰腺自噬的影响及其机制。方法 将28只大鼠随机分为正常妊娠组(A组)、GDM组(B组)、GDM褪黑素低剂量组(C组)及GDM褪黑素高剂量组(D组),每组7只。检测各组大鼠血清中葡萄糖、胰岛素及糖化血红蛋白水平,并制作各组大鼠胰腺组织病理切片,HE染色观察各组大鼠胰腺组织病理变化,Tunel染色检测胰腺细胞凋亡情况,透射电镜检测胰腺组织自噬情况,免疫印迹法检测胰腺组织中自噬相关蛋白微管相关蛋白1轻链3(LC3)、Beclin1、磷酸化磷脂酰肌醇-3-激酶(p-PI3K)、磷脂酰肌醇-3-激酶(PI3K)、磷酸化蛋白激酶B(p-AKT)、蛋白激酶B(AKT)、磷酸化哺乳动物雷帕霉素靶蛋白(p-mTOR)和哺乳动物雷帕霉素靶蛋白(mTOR)的表达水平。结果 各组大鼠血清葡萄糖、胰岛素及糖化血红蛋白水平比较均有显著差异(F=52.26~66.73,P<0.05);与A组相比,B组大鼠血清葡萄糖、胰岛素及糖化血红蛋白水平显著升高(t=-14.45~7.23,P<0.05);与B组相比,C组、D组大鼠血清葡萄糖、胰岛素及糖化血红蛋白含量显著降低(t=-5.32~10.02,P<0.05)。HE染色、Tunel染色及透射电镜观察结果显示,与A组大鼠相比,B组大鼠胰腺组织病理损伤及细胞凋亡加重,胰腺组织自噬水平增加;与B组大鼠相比,C、D组大鼠胰腺组织病理损伤及细胞凋亡减轻,胰腺组织自噬水平降低。免疫印迹法结果显示,各组大鼠胰腺组织中LC3Ⅱ/LC3Ⅰ、p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR水平以及Beclin1相对表达量比较差异均具有显著性(F=27.68~67.72,P<0.05);与A组大鼠相比,B组大鼠胰腺组织LC3Ⅱ/LC3Ⅰ水平和Beclin1相对表达量均显著升高(t=-8.13~-6.83,P<0.05),p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR水平显著下降(t=8.83~14.05,P<0.05);与B组大鼠相比,C、D组大鼠胰腺组织LC3Ⅱ/LC3Ⅰ水平和Beclin1相对表达量均显著下降(t=3.36~6.95,P<0.05),p-PI3K/PI3K、p-AKT/AKT、p-mObjective To investigate the effect of melatonin on pancreatic autophagy in rats with gestational diabetes mellitus(GDM)and the underlying mechanism.Methods Twenty-eight rats were randomly divided into A group(rats with normal pregnancy),B group(rats with GDM),C group(low-dose melatonin administration for rats with GDM),and D group(high-dose melatonin administration for rats with GDM),with seven rats in each group.All the groups were examined for the se-rum levels of glucose,insulin,and glycosylated hemoglobin.Pancreatic tissue slices were prepared to examine the pathological changes of the pancreas with hematoxylin-eosin(HE)staining,the apoptosis of pancreatic cells with TUNEL staining,and the autophagy of the pancreatic tissue with transmission electron microscopy.Western blot was used to measure the expression of autop-hagy-related proteins in the pancreatic tissue:microtubule-associated protein 1 light chain 3(LC3),Beclin1,phosphorylated phosphatidylinositol 3-kinase(p-PI3K),PI3K,phosphorylated protein kinase B(p-AKT),AKT,phosphorylated mammalian target of rapamycin(p-mTOR),and mTOR.Results There were significant differences in serum glucose,insulin,and glycosylated hemoglobin levels between different groups(F=52.26-66.73,P<0.05).Compared with those of group A,the levels of serum glucose,insulin,and glycosylated hemoglobin in group B were significantly increased(t=-14.45-7.23,P<0.05).Compared with group B,groups C and D showed significantly decreased serum glucose,insulin,and glycosylated hemoglobin levels(t=-5.32-10.02,P<0.05).The results of HE staining,TUNEL staining,and transmission electron microscopy revealed that compared with group A,group B showed increased levels of pathological damage,apoptosis,and autophagy in the pancreatic tissue;groups C and D showed reduced pathological damage,apoptosis,and autophagy in the pancreatic tissue than group B.Western blot detected significant differences in the levels of LC3Ⅱ/LC3Ⅰ,p-PI3K/PI3K,p-AKT/AKT,and p-mTOR/mTOR and the relative expression of Beclin1 in th

关 键 词：糖尿病 妊娠 褪黑激素 胰腺 磷酸肌醇3-激酶类 原癌基因蛋白质c-akt 自噬 大鼠 

分 类 号：R714.256[医药卫生—妇产科学]