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
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 stay-ing time and the percentage of times entering the open arm of rats;the gastric emptying rate and small intestinal pro-pulsion 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 expres-sion 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 per-centage 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 signifi-cantly decreased(P＜0.05 or P＜0.01);the ultrastructure of ENS-PDGFRα+cell-SMC network was damaged with-out 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 propul-sion rate,and the gastric empting rate of rats in medium-and high-dose Shuwei Decoction groups increased after ga-vage;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(P＜0.05 or P＜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(P＜0.05 or P＜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.
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