左归丸对GDM模型母鼠子鼠胰岛功能的影响  

Effect of Zuoguiwan on Pancreatic Islet Function in Offspring of Maternal Rats with GDM

作　　者：谭贝西 赵乐 冯忆 王恬恬 王超群 王永辉 TAN Beixi;ZHAO Le;FENG Yi;WANG Tiantian;WANG Chaoqun;WANG Yonghui(Shanxi University of Chinese Medicine,Jinzhong 030619,China)

机构地区：[1]山西中医药大学,山西晋中030619

出　　处：《中国实验方剂学杂志》2023年第17期1-8,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金面上项目(82274378)。

摘　　要：目的:探讨左归丸对妊娠期糖尿病(GDM)模型母鼠子鼠胰岛功能的影响,并基于胰腺产后增殖过程探讨左归丸改善子鼠胰岛功能的作用机制。方法:选取血糖正常的健康雌性SD大鼠与雄性大鼠按照2∶1比例同笼,以阴道栓或阴道涂片为依据,选取受孕雌性大鼠将其分为正常组、模型组、胰岛素组(地特胰岛素,20 U·kg^(-1))和左归丸低、高剂量组(1.89、3.78 g·kg^(-1))。除正常组外,用链脲佐菌素复制GDM大鼠模型,通过对母鼠血糖检测判断模型是否建立,除正常组和模型组外,其余组均每日给药。于子鼠出生后21 d时,每组检测6只子鼠空腹血糖值(FBG)与血清中空腹胰岛素(FINS),计算胰岛素抵抗指数(HOMA-IR),每组另取12只子鼠进行口服糖耐量(OGTT)检测;出生22 d(B22d)腹主动脉取血,利用酶联免疫吸附测定法(ELISA)检测各组子鼠血清中胰岛素(INS)、胰高血糖素(GC)、胰多肽(PPY)、生长抑素(SS)观察胰岛各细胞所表达激素的水平,利用苏木素-伊红(HE)染色观察各组子鼠胰腺组织病理变化;利用免疫荧光法(IF)观察子鼠胰岛面积与结构;且通过蛋白免疫印迹法(Western blot)检测子鼠胰腺β细胞发育成熟、功能表达密切相关的关键蛋白胰腺十二指肠同源框蛋白1(Pdx1)、Nkx6.1、葡萄糖运载体2(Glut2)表达。结果:与正常组比较,模型组子鼠FBG水平、FINS水平及HOMA-IR均显著升高(P<0.01);与模型组比较,胰岛素组FBG水平及HOMA-IR显著降低(P<0.01),左归丸低剂量组FBG水平明显降低(P<0.05),左归丸高剂量组FBG水平、FINS水平及HOMA-IR均显著降低(P<0.01)。与正常组比较,模型组大鼠OGTT 60 min血糖值、曲线下面积(AUC)指数明显升高(P<0.05,P<0.01);与模型组比较,左归丸高剂量组子鼠OGTT血糖值在60 min、AUC指数明显降低(P<0.05,P<0.01)。胰腺组织HE染色观察,与正常组比较,模型组胰岛数量减少,腺泡细胞排列疏松;与模型组比较,给药各组胰岛数量�Objective:To investigate the effect of Zuoguiwan on pancreatic islet function in offspring of gestational diabetes mellitus(GDM)maternal rat model and explore the mechanisms of Zuoguiwan in improving pancreatic islet function based on postpartum pancreatic regeneration.Method:Healthy female SD rats with normal blood glucose levels were paired with male rats in a 2∶1 ratio and housed together.Pregnancy was confirmed based on vaginal plugs or vaginal smears.The pregnant rats were divided into the following groups:normal group,model group,insulin group(insulin Detemir,20 U·kg^(-1)),low-dose Zuoguiwan group(1.89 g·kg^(-1)),and high-dose Zuoguiwan group(3.78 g·kg^(-1)).The GDM rat model was induced using streptozotocin in rats except for those in the normal group.The model was confirmed by blood glucose testing in the maternal rats.Except for the normal and model groups,the other groups received daily administration of corresponding treatments.At 21 days after birth,fasting blood glucose(FBG)and fasting serum insulin(FINS)levels were measured in 6 offspring from each group.The homeostasis model assessment of insulin resistance(HOMA-IR)was calculated,and an oral glucose tolerance test(OGTT)was performed on additional 12 offspring from each group.Blood samples were taken from the abdominal aorta of the offspring at postnatal day 22,and enzyme-linked immunosorbent assay(ELISA)was used to measure insulin,glucagon(GC),pancreatic polypeptide(PPY),and somatostatin(SS)levels in the serum.Hematoxylin-eosin(HE)staining was performed to observe pathological changes in the pancreatic tissue of the offspring.Immunofluorescence(IF)was used to observe the area and structure of the pancreatic islets.Western blot was used to detect the expression of key proteins involved in the development and functional expression of pancreaticβ-cells,namely pancreatic and duodenal homeobox factor 1(Pdx1),Nkx6.1,and Glucose transporter 2(Glut2).Result:Compared with the normal group,the model group showed significant increases in FBG and FINS le

关 键 词：妊娠期糖尿病 胰腺发育 胰岛功能 左归丸 

分 类 号：R2-0[医药卫生—中医学] R22R285.5R289R33