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

TAN Beixi; ZHAO Le; FENG Yi; WANG Tiantian; WANG Chaoqun; WANG Yonghui

doi:10.13422/j.cnki.syfjx.20230604

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

更新时间：2023-08-07

- Effect of Zuoguiwan on Pancreatic Islet Function in Offspring of Maternal Rats with GDM
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 17, Pages: 1-8(2023)
- 作者机构：
  
  山西中医药大学，山西晋中 030619
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20230604
  CLC： R2-0;R22;R285.5;R289;R33
- Published：05 September 2023，
  
  Published Online：16 February 2023，
  
  Received：03 November 2022，
- 稿件说明：
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谭贝西,赵乐,冯忆等.左归丸对GDM模型母鼠子鼠胰岛功能的影响[J].中国实验方剂学杂志,2023,29(17):1-8.

TAN Beixi,ZHAO Le,FENG Yi,et al.Effect of Zuoguiwan on Pancreatic Islet Function in Offspring of Maternal Rats with GDM[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(17):1-8.
谭贝西,赵乐,冯忆等.左归丸对GDM模型母鼠子鼠胰岛功能的影响[J].中国实验方剂学杂志,2023,29(17):1-8. DOI： 10.13422/j.cnki.syfjx.20230604.

TAN Beixi,ZHAO Le,FENG Yi,et al.Effect of Zuoguiwan on Pancreatic Islet Function in Offspring of Maternal Rats with GDM[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(17):1-8. DOI： 10.13422/j.cnki.syfjx.20230604.

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

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（

0.01）；与模型组比较，胰岛素组FBG水平及HOMA-IR显著降低（

0.01），左归丸低剂量组FBG水平明显降低（

0.05），左归丸高剂量组FBG水平、FINS水平及HOMA-IR均显著降低（

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（

0.05，

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（

0.05，

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，给药各组胰岛数量增加，腺泡细胞排列紧密。与正常组比较，模型组子鼠血清中INS、GC、PPY、SS含量均显著升高（

0.01）；与模型组比较，左归丸低、高剂量组及胰岛素组子鼠血清中INS、GC、PPY、SS含量均明显降低（

0.05，

0.01）。免疫荧光结果显示，与正常组比较，模型组INS阳性率明显降低（

0.05）；与模型组比较，左归丸低高剂量组明显增高（

0.05）；GC阳性率各组间差异无统计学意义；单个胰岛中INS与GC所占比例中，与正常组比较，模型组INS所占比例显著降低（

0.01），GC所占比例显著增高（

0.01）；与模型组比较，左归丸低、高剂量组INS所占比例显著增高（

0.01），GC所占比例显著降低（

0.01）。与正常组比较，模型组GDM子鼠胰腺组织中Pdx1、Nkx6.1、Glut2蛋白表达含量均明显降低（

0.05）；与模型组比较，胰岛素组、左归丸低剂量组GDM子鼠胰腺组织中Pdx1、Nkx6.1蛋白表达含量均明显增高（

0.05），左归丸低、高剂量组Glut2蛋白表达含量均明显增高（

0.05）。

结论

左归丸可促进GDM模型大鼠子鼠胰岛发育，改善胰岛形态及功能，缓解胰岛素抵抗表现，其作用机制可能与调节子代胰腺组织中Pdx1、Nkx6.1、Glut2蛋白表达有关。

Abstract

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 pancreatic

-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 levels， and HOMA-IR （

0.01）. Compared with the model group， the insulin group showed significant decreases in FBG levels and HOMA-IR （

0.01）， the low-dose Zuoguiwan group showed a significant decrease in FBG levels （

0.05）， and the high-dose Zuoguiwan group showed significant decreases in FBG and FINS levels， and HOMA-IR （

0.01）. Compared with the normal group， the model group showed significant increases in OGTT 60-min blood glucose levels and AUC index （

0.05，

0.01）. Compared with the model group， the high-dose Zuoguiwan group showed significant decreases in OGTT60-min blood glucose levels and area under the curve（AUC） index （

0.05，

0.01）. HE staining of pancreatic tissue showed that compared with the normal group， the model group had a reduced number of islets and a loose arrangement of acinar cells. Compared with the model group， the groups with drug treatment showed increased number of islets and a compact arrangement of acinar cells. Compared with the normal group， the model group had significantly increased levels of insulin， GC， PPY， and SS in the serum （

0.01）. Compared with the model group， the low-dose and high-dose Zuoguiwan groups and the insulin group showed significantly decreased serum levels of insulin， GC， PPY， and SS （

0.05，

0.01）. IF results showed that compared with the normal group， the model group had a significantly lower positive rate of insulin （

0.05）. Compared with the model group， the low-dose and high-dose Zuoguiwan groups showed a significant increase in the positive rate of insulin （

0.05）. There was no significant difference in the positive rate of GC among the groups. In terms of the proportion of insulin and GC in individual islets， compared with the normal group， the model group showed a significant decrease in the proportion of insulin （

0.01） and a significant increase in the proportion of GC （

0.01）. Compared with the model group， the low-dose and high-dose Zuoguiwan groups showed significantly increased proportion of insulin （

0.01） and significantly decreased proportion of GC （

0.01）. Compared with the normal group， the model group showed significantly decreased expression levels of Pdx1， Nkx6.1， and Glut2 proteins in the pancreatic tissue of GDM offspring （

0.05）. Compared with the model group， the insulin group and the low-dose Zuoguiwan group showed significant increases in the expression levels of Pdx1 and Nkx6.1 proteins in the pancreatic tissue of GDM offspring （

0.05）， and the low-dose and high-dose Zuoguiwan groups showed significant increases in the expression levels of Glut2 protein （

0.05）.

Conclusion

Zuoguiwan can promote pancreatic islet development in offspring of GDM maternal rat model， improve pancreatic islet morphology and function， and alleviate insulin resistance. Its mechanism of action may be related to the regulation of Pdx1， Nkx6.1， and Glut2 protein expression in the pancreatic tissue of offspring.

关键词

妊娠期糖尿病胰腺发育胰岛功能左归丸

Keywords

gestational diabetes mellituspancreas developmentpancreatic islet functionZuoguiwan

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