The Pharmacodynamics And Mechanism Of Different Extraction Of Poria Cocos On Tonifying Spleen

Background: Poria cocos(Schw.)Wolf.Which is a dried sclerotium of the Polyporaceae fungus,and it is a commonly used Chinese medicine.It is first recorded in the Shennong Herbal Classic,and it is sweet,and mild;It belongs to the heart,lungs,spleen,and kidneys in Chinese medicine,the main function is diuresis,tonifying spleen and calming nerves.Triterpenoids and polysaccharides are the main components of Poria cocos,in addition to other components such as steroids,amino acids,volatile oils,choline,histidine and potassium salts." The spleen provides the material basis for the acquired constitution " indicates that the "spleen" plays an important role in traditional Chinese medicine ".Loss of spleen transport is often manifested as spleen deficiency.Spleen Qi Deficiency Syndrome refers to qi deficiency and spleen loss of healthy transportation.Common symptoms include reduced appetite,abdominal distension,loose stool,fatigue,weakness of limbs and weak pulse.Western medicine believes that the spleen is the human immune organ and mainly plays a role in regulating immunity,while Chinese medicine believes that the spleen is related to the human digestive system,nervous system,and immune system.At present,the traditional efficacy of Poria cocos and the mechanism of action are not clear.This study is based on the theory of tonifying spleen of Poria cocos,and comprehensively and accurately interprets the material basis and mechanism under the guidance of Chinese medicine theory.Objective: This article aims to establish a model of spleen deficiency in rats by using a combination of dietary insufficiency and overwork,to clarify the pharmacodynamics and action mechanism of different extracts of Poria cocos on spleen deficiency.The mechanism of action provides a scientific basis.Methods: A rat model of spleen deficiency was constructed by using eating disorder and overwork.The Model group was fed every other day.The rats swim to exhaustion every day,the Control group was fed normally without swimming,for 6 weeks.After the modeling was completed,the positive Control group were administrated daily with Buzhong Yiqi Pill solution,PCD group was administrated with Poria cocos decoction daily,and PCT group,PCWP group and PCAP group were administrated corresponding extraction daily,PCPO group was administrated with Poria cocos power daily,for 2 weeks.At the end of the experiment,a 3% D-xylose solution was administered to 10 m L/kg,and then serum,heart,liver,spleen,lung,kidney,stomach,small intestine,and thymus of all rats were taken out,and organ coefficients were calculated.The D-xylose,ALB,TP,ACH in rat serum were measured by biochemical kit;The GAS,MTL,CCKAR,VIP,EGF,TGF-?1,IL-17,IL-2,IFN-?,IL-4,IL-1?,IL-6 and TNF-? in rat serum were measured by ELISA kit;H&E staining is used to determine the pathophysiological changes of stomach and small intestine;immunohistochemical staining was used to detect the expression of Ghrelin and EGFR in gastric,MUC2 in small intestine tissue,and AQP1 and AQP2 in kidney;Western blot was used to detect T-bet,GATA3,ROR-?t,and Foxp3 expression in spleen.Results: During the modeling period,from the 6th day,the rats swimming endurance decreased,hair was dull and caducous,and stool became dirty.From the 8th day,the diet decreased,and the weight was significantly lower than Control group,loose stool and fatigue appeared.During the administration period,from the 3rd day,BZYQP group,PCD group and PCT group showed an increase in swimming time,a decrease in hair loss,a softer stool,and a faster weight gain.From the 7th day,the weight of the rats in each group was significantly higher than the Model group.The swimming time increased,the hair became shiny,and the mental state improved.By the 12 th day,the stool in the administration group was basically normal.The results of the biochemical kit showed that compared with the Control group,the contents of D-xylose,ACH,ALB and TP in the Model group were significantly decreased(p<0.01).Compared with the Model group,the contents of D-xylose,ACH and TP in the PCT group were significantly increased(p<0.05),and the contents of D-xylose and ACH in the PCWP group were significantly increased(p<0.05).The contents of D-xylose,ACH,ALB and TP in PCD group in PCAP group and PCPO group were significantly increased(p<0.01 or p<0.05).ELISA results showed that compared with the Control group,the contents of GAS,MTL,SP,IL-2,IL-4 and TGF-?1 in the Model group were significantly reduced(p<0.01 or p<0.05),while the contents of VIP,CCKAR,5-HT,CGRP,EGF,IFN-?,IL-1,IL-6,IL-17 and TNF-? were significantly increased(p<0.01 or p<0.05).Compared with the Model group,the contents of GAS,MTL,SP,EGF,IL-2 and IL-4 in PCD group were significantly increased(p<0.01 or p<0.05),the contents of VIP,CCKAR,5-HT,CGRP,IFN-?,IL-1,IL-6,IL-17 and TNF-? were significantly decreased(p<0.01 or p<0.05).The contents of GAS,SP,EGF and IL-4 in PCT group were significantly increased(p<0.01 orp<0.05),the contents of CCKAR,5-HT,CGRP,IL-1,IL-6,IL-17,TNF-? were significantly reduced(p<0.01 or p<0.05).The contents of GAS,SP,EGF,IL-2,IL-4,TGF-?1 in PCWP group were significantly increased(p<0.01 or p<0.05),the contents of VIP,CCKAR,5-HT,CGRP,IFN-?,IL-1,IL-17,TNF-? were significantly decreased(p<0.01 or p<0.05).The contents of EGF,IL-4,TGF-?1 in the PCAP group were significantly increased(p<0.01 or p<0.05),the contents of 5-HT,IFN-?,IL-1,IL-6,IL-17 and TNF-? were significantly decreased(p<0.01 or p<0.05).The contents of GAS,EGF,IL-2,IL-4 and TGF-?1 in the PCPO group were significantly increased(p<0.01 or p<0.05),and the contents of VIP,5-HT,IL-1,IL-6 and IL-17 were significantly decreased(p<0.01 or p<0.05);H&E staining results showed that the gastric tissue in the Control group had normal morphology and no obvious pathological changes.The small intestinal mucosal epithelium was intact with not fall off.In the Model group,the gastric mucosal epithelial cells were necrotic,the tissue morphology was destroyed,the small intestinal villi structure was incomplete,and the mucosa epithelial cells were obviously detached.After administration,each administration group could improve gastric tissue lesions in rats with spleen deficiency.The PCT group and PCAP group were significantly improved the small intestinal mucosa.Immunohistochemical staining results showed that compared with the Control group,in Model group,the expression of Ghrelin,EGFR,MUC2 were decreased,and the expression of AQP1,AQP2 were increased.Compared with Model group,the expression of Ghrelin,EGFR and MUC2 in PCD group was increased,the expression of AQP1 and AQP2 were decreased.The expression of Ghrelin,EGFR and MUC2 in PCT group were increased,the expression of AQP1 and AQP2 were decreased.The expression of MUC2 in PCWP group were increased,the expression of AQP1 and AQP2 were decreased.The expression of Ghrelin,EGFR and MUC2 in PCAP group were increased,the expression of AQP2 were decreased.The expression of Ghrelin and MUC2 in PCPO group were increased,the expression of AQP1 and AQP2 were decreased;Western blot results showed that compared with the Control group,the expressions of T-bet and ROR-?t in the Model group were significantly increased(p<0.01),the expression of GATA3 and Foxp3 was significantly reduced(p<0.01).Compared with the Model group,the expression of T-bet in PCD group was significantly reduced(p<0.05),the expression of GATA3 was significantly increased(p<0.01).The expression of ROR-?t in PCT group was significantly reduced(p<0.01),the expression of GATA3 and Foxp3 were significantly increased(p<0.01 or p<0.05).In PCWP group and PCAP group the expression of T-bet and ROR-?t were significantly reduced(p<0.01),the expression of GATA3 and Foxp3 were significantly increased(p<0.01).The expression of ROR-?t in PCPO group was significantly reduced(p<0.01),and the expression of GATA3 and Foxp3 was significantly increased(p<0.01).Conclusion: Different extraction of Poria cocos have good therapeutic effects on spleen deficiency,but their mechanisms of action are different.PCT,PCWP and PCAP can protect gastrointestinal mucosa by regulating the contents of serum EGF and MUC2 in the small intestine,reduce the contents of AQP1 or AQP2 in the kidney to improve water metabolism disorders,and regulate the imbalance of Th17/Treg cells to enhance immunity.Among them,PCT and PCAP can promote the repair of gastric mucosa by regulating the expression of EGFR.PCT and PCWP can also improve gastrointestinal function by regulating the brain intestinal peptide.PCWP and PCAP can also enhance immunity by regulating the imbalance of Th1/Th2 cells.