Exploring the Effect of <italic style="font-style: italic">Shuwei Decoction</italic> （舒胃汤） for Rats with Functional Dyspepsia of Liver-Depression and Spleen-Deficiency Syndrome Model on the Ultrastructure of ENS-PDGFRα<sup>+</sup> Cell-SMC Network in Gastric Sinusoidal Tissues

LIANG Junyao; GUO Xuan; ZHANG Dexu; ZHOU Man; XIAO Jie; Tang Chusen; Feng Hui; XU Yin

doi:10.13288/j.11-2166/r.2024.12.013

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Exploring the Effect of Shuwei Decoction （舒胃汤） for Rats with Functional Dyspepsia of Liver-Depression and Spleen-Deficiency Syndrome Model on the Ultrastructure of ENS-PDGFRα⁺ Cell-SMC Network in Gastric Sinusoidal Tissues

更新时间：2024-06-12

- Exploring the Effect of Shuwei Decoction （舒胃汤） for Rats with Functional Dyspepsia of Liver-Depression and Spleen-Deficiency Syndrome Model on the Ultrastructure of ENS-PDGFRα⁺ Cell-SMC Network in Gastric Sinusoidal Tissues
- Journal of Traditional Chinese Medicine Vol. 65, Issue 12, Pages: 1269-1278(2024)
- 作者机构：
  
  湖南中医药大学第一附属医院，湖南省长沙市雨花区韶山中路95号，410000
- 作者简介：
- 基金信息：
- DOI：10.13288/j.11-2166/r.2024.12.013
  CLC：
- Received：28 November 2023，
  
  Revised：2024-02-25，
  
  Published：17 June 2024
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梁俊尧,郭璇,张德旭等.舒胃汤对功能性消化不良肝郁脾虚证模型大鼠胃窦组织中ENS-PDGFRα⁺细胞-SMC网络超微结构的影响[J].中医杂志,2024,65(12):1269-1278. DOI: 10.13288/j.11-2166/r.2024.12.013.

LIANG Junyao,GUO Xuan,ZHANG Dexu,et al.Exploring the Effect of Shuwei Decoction （舒胃汤） for Rats with Functional Dyspepsia of Liver-Depression and Spleen-Deficiency Syndrome Model on the Ultrastructure of ENS-PDGFRα⁺ Cell-SMC Network in Gastric Sinusoidal Tissues[J].Journal of Traditional Chinese Medicine,2024,65(12):1269-1278. DOI: 10.13288/j.11-2166/r.2024.12.013.
梁俊尧,郭璇,张德旭等.舒胃汤对功能性消化不良肝郁脾虚证模型大鼠胃窦组织中ENS-PDGFRα⁺细胞-SMC网络超微结构的影响[J].中医杂志,2024,65(12):1269-1278. DOI: 10.13288/j.11-2166/r.2024.12.013. DOI：

LIANG Junyao,GUO Xuan,ZHANG Dexu,et al.Exploring the Effect of Shuwei Decoction （舒胃汤） for Rats with Functional Dyspepsia of Liver-Depression and Spleen-Deficiency Syndrome Model on the Ultrastructure of ENS-PDGFRα⁺ Cell-SMC Network in Gastric Sinusoidal Tissues[J].Journal of Traditional Chinese Medicine,2024,65(12):1269-1278. DOI: 10.13288/j.11-2166/r.2024.12.013. DOI：

摘要

目的

基于肠神经系统（ENS）-血小板衍生的生长因子受体α阳性（PDGFRα

）细胞-平滑肌细胞（SMC）网络超微结构探讨舒胃汤治疗功能性消化不良（FD）肝郁脾虚证的可能作用机制。

方法

将50只SD大鼠随机分为空白组、模型组及舒胃汤低、中、高剂量组，每组10只。空白组不予造模，其余大鼠采用改良复合病因造模法（慢性束缚应激+夹尾激怒+过度疲劳+饮食失节）建立FD肝郁脾虚证大鼠模型，持续21天。造模完成后舒胃汤低、中、高剂量组大鼠分别给予舒胃汤药液3.78、7.56、15.12 g/（kg·d）灌胃，空白组和模型组大鼠给予生理盐水10 ml/（kg·d）灌胃，均每日灌胃1次，持续14天。实验结束后比较各组大鼠灌胃前后体质量增长值，进行高架十字迷宫实验观察大鼠开臂停留时间百分比和进入开臂次数百分比；黑色半固体糊灌胃法检测大鼠胃排空率、小肠推进率；HE染色法观察胃窦组织病理情况；Western Blot法和RT-qPCR法分别检测胃窦组织血小板衍生的生长因子受体α（PDGFRα）、小电导钙激活钾通道3（Sk3）、嘌呤能P2Y1受体（P2Y1）蛋白及mRNA表达；免疫

荧光双染检测胃窦组织中PDGFRα与酪氨酸激酶受体（C-kit）、蛋白基因产物9.5（PGP9.5）、Sk3、平滑肌细胞（SMC）的共定位情况；透射电镜观察ENS-PDGFRα

细胞-SMC网络的超微结构。

结果

HE染色结果显示，各组大鼠胃组织黏膜皱襞清晰，未见出血、溃疡、肿胀等器质性病变，模型组及舒胃汤低剂量组胃窦组织固有层内部分区域胃腺排列较为紊乱，舒胃汤中、高剂量组固有层内胃腺排列较为整齐，病变程度轻于模型组及舒胃汤低剂量组。与空白组比较，模型组大鼠灌胃前后体质量增长值、开臂停留时间百分比、进入开臂次数百分比、小肠推进率、胃排空率均明显降低，胃窦组织中PDGFRα、Sk3、P2Y1蛋白及mRNA表达降低，PDGFRα与C-kit、Sk3、SMC、PGP9.5共定位表达明显降低（

＜0.05或

＜0.01），ENS-PDGFRα

细胞-SMC网络超微结构破坏，未见缝隙连接。与模型组和舒胃汤低剂量组比较，舒胃汤中、高剂量组大鼠灌胃前后体质量增长值、开臂停留时间百分比、进入开臂次数百分比、小肠推进率、胃排空率均升高，胃窦组织中PDGFRα、Sk3、P2Y1蛋白及mRNA表达升高，舒胃汤高剂量组PDGFRα与C-kit、Sk3、SMC、PGP9.5共定位表达明显升高（

＜0.05或

＜0.01），ENS-PDGFRα

细胞-SMC网络超微结构完整。与舒胃汤中剂量组比较，舒胃汤高剂量组PDGFRα、Sk3、P2Y1蛋白表达，PDGFRα、Sk3 mRNA表达，以及PDGFRα与C-kit、Sk3、SMC、PGP9.5共定位表达均升高（

＜0.05或

＜0.01）。

结论

舒胃汤可有效改善FD肝郁脾虚证模型大鼠胃窦组织的病理损伤，提高大鼠胃排空率、小肠推进率，其可能作用机制与保护ENS-PDGFRα

细胞-SMC网络超微结构完整性有关。

Abstract

Objective

Based on the ultrastructure of enteric nervous system （ENS）-platelet-derived growth factor receptor α positive （PDGFRα

） cell - smooth muscle cell （SMC） network， the effect and mechanism of

Shuwei Decoction

（舒胃汤） for rats with functional dyspepsia （FD） of liver-depression and spleen-deficiency syndrome were explored.

Methods

Fifty SD rats were divided into blank group， model group， low-， medium- and high-dose

Shuwei Decoction

groups randomly， with 10 rats in each group. The blank group was not set up， and the other 4 groups were set up with the modified composite cause （chronic restraint stress + tail irritation + excessive fatigue + diet disorder） for 21 days， resulting in FD model rats with liver-depression and spleen-deficiency syndrome. After successful modelling， rats in the low-， medium- and high-dose groups were given 3.78， 7.56 and 15.12 g/（kg·d） of

Shuwei Decoction

by gavage， respectively， while rats in the blank group and the model group were given 10 ml/（kg·d） of saline by gavage； all of them were given gavage once a day for 14 days. Body mass increase were collected before and after gavage， and compared after the experiment； the elevated cross maze experiment was conducted to observe the percentage of staying time and the percentage of times entering the open arm of rats； the gastric emptying rate and small intestinal propulsion rate were measured by black semi-solid paste； HE staining was used to observe histopathology of the gastric antrum； the Western Blotting and RT-qPCR were used to detect the expression levels of PDGFRα， small conductance calcium activated potassium （SK3）， and purinergic G protein-coupled receptor P2Y1 （P2Y1 R）， and mRNA expression in gastric antrum； immunofluorescence double staining was used to detect the co-localization of PDGFRα with receptor tyrosine kinase （C-kit）， protein gene product 9.5 （PGP9.5）， Sk3， and smooth muscle cells （SMCs） in gastric sinusoidal tissues； and transmission electron microscopy was used to observe the ultrastructure of the ENS-PDGFRα

cell-SMC network.

Results

HE staining results showed that the mucosal folds of gastric tissue were clear in all groups， no organic lesions such as bleeding， ulceration and swelling were observed， and the arrangement of gastric glands in the lamina propria of gastric sinusoidal tissues in model group and low-dose

Shuwei Decoction

group was disordered. The arrangement of gastric glands in lamina propria in

medium- and high-dose

Shuwei Decoction

groups was more orderly， and the lesion degree was less than that in model group and low-dose

Shuwei Decoction

group. Compared with the blank group， the growth value of body mass， the percentage of open arm retention time， the percentage of times entering the open arm， the small intestine propulsion rate， and

the gastric empty rate of rats in the model group significantly decreased after gavage； the expressions of PDGFRα， Sk3， P2Y1 protein and mRNA in gastric sinusoidal tissues decreased； the co-localization expression of PDGFRα with C-kit， Sk3， SMC and PGP9.5 significantly decreased （

＜0.05 or

＜0.01）； the ultrastructure of ENS-PDGFRα

cell-SMC network was damaged without gap junction. Compared with model group and low-dose

Shuwei Decoction

group， the growth value of body mass， the percentage of open arm retention time， the percentage of times entering the open arm， the small intestine propulsion rate， and the gastric empting rate of rats in medium- and high-dose

Shuwei Decoction

groups increased after gavage； the expressions of PDGFRα， Sk3， P2Y1 protein and mRNA in gastric sinusoidal tissues increased； the co-localization expression of PDGFRα with C-kit， Sk3， SMC and PGP9.5 significantly increased （

＜0.05 or

＜0.01）； the ultrastructure of ENS-PDGFRα

cell-SMC network was intact. Compared with medium-dose

Shuwei Decoction

group， the protein expressions of PDGFRα， Sk3 and P2Y1， the mRNA expressions of PDGFRα and Sk3， and the co-localization expressions of PDGFRα and C-kit， Sk3， SMC， and PGP9.5 in high-dose

Shuwei Decoction

group increased （

＜0.05 or

＜0.01）.

Conclusion

Shuwei Decoction

can improve the pathological lesion of the gastric antrum in the FD model rats with liver-depression and spleen-deficiency syndrome and increase the gastric emptying rate as well as the small intestinal propusion rate. The mechanism of

Shuwei Decoction

on FD rats with liver-depression and spleen-deficiency syndrome is related to the protection of the ultrastructural integrity of ENS-PDGFRα

cell-SMC network.

关键词

Keywords

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⁰

Exploring the Effect of Shuwei Decoction （舒胃汤） for Rats with Functional Dyspepsia of Liver-Depression and Spleen-Deficiency Syndrome Model on the Ultrastructure of ENS-PDGFRα+ Cell-SMC Network in Gastric Sinusoidal Tissues

DOI：10.13288/j.11-2166/r.2024.12.013

摘要

Abstract

关键词

Keywords

references

Exploring the Effect of Shuwei Decoction （舒胃汤） for Rats with Functional Dyspepsia of Liver-Depression and Spleen-Deficiency Syndrome Model on the Ultrastructure of ENS-PDGFRα⁺ Cell-SMC Network in Gastric Sinusoidal Tissues