From The Four Gentlemen's Soup To Promote The Healing Of Spleen-deficiency Gastric Ulcer, Explore The Mechanism Of "the Spleen Main Guard" In Neijing

Objective:The syndrome of spleen deficiency is a common clinical syndrome.Clinical data show that the deficiency of spleen qi is related to the occurrence of gastric ulcer and may be through the impairment of the protective mechanism of gastric mucosa.The idea of "spleen governs protection" originates from the Yellow Emperor 's Inner Classic.However,at present there is little explanation or discussion to its connotation.Besides,Sijunzi decoction(SJZD)exert a good therapeutic effect on gastric ulcer in clinical practice,but there is no systematic evaluation on its efficacy.Additionally,the fact that clinical application of fortifying the spleen and boosting qi method,represented by SJZD,can promote the healing of peptic ulcer with spleen deficiency syndrome is one embodiment of the thought of "spleen governs protection",but there is limited research on its mechanism.HGF and its receptor c-Met can promote the proliferation and migration of injured gastric mucosal epithelial cells,promote mucosal repair after gastric mucosal injury,and promote ulcer healing through PI3K/Akt pathway in the mucosal repair after gastric ulcer injury.However,there is little study on the involvement of HGF,c-Met and PI3K/Akt signaling pathways in the treatment of gastric ulcer by SJZD.What's more,the gut axis is the key factor to maintain the integrity of gastrointestinal mucosa,to enhance the defensive ability of gastrointestinal mucosa and to promote the repair of gastrointestinal mucosa.Whether SJZD promotes the spleen-deficiency ulcer healing through the mechanism of central system,especially EGFR,ERK and SSTR and TFF3 in the brain remains to be explored.Therefore,this study aims to elucidate "spleen governs protection"theory and to explore its mechanism through investigation of mechanism of SJZD promoting the spleen-deficiency gastric ulcer healing.Methods:This study is divided into three parts.The first part uses document collation method to study and analyze the theory of "spleen governs protection" in Yellow Emperor's Inner Classic.The second part carries an evidence-based systematic review and meta-analysis to appraise SJZD treatment on peptic ulcer based on clinical randomized controlled researches.The third part experimentally investigates the influence of SJZD on rat model of spleen deficiency gastric ulcer and its mechanisms including the mucous membrane healing quality,gastrointestinal mucosa protective mechanism,signaling pathway and central mechanism.Specifically,the methods employed include:(1)a combination method of breaking stagnant qi and depleting qi,irregular eating and acetic acid to duplicate rat model of spleen deficiency gastric ulcer;(2)The immunohistochemical method to detect the changes of HGF and c-Met in the gastral mucosa and EGFR,ERK1/2,SSTR2 and TFF3 in hypothalamus and hippocampus;(3)Western-blot method to observe gastric PI3K,Akt changes;and(4)HE staining method to observe the histological maturity of regenerated mucosa.Results:(1)Theoretical research holds that the connotation of "spleen governs protection"is that the spleen can protect the body from pathogens,expel pathogens after catching the disease,promote the disease healing,prevent disease transferring to other zang and fu organs and meridians and collaterals and prevent disease relapse.All these originate from its function of "governing transportation and transformation" and thereby being "the source of qi and blood"to the nourish zang and fu organs,limbs and orifices.(2)Systematic review and meta-analyses found that,fourteen trails fulfilled the selection criteria.Of them,favorable effect of modified SJZD were showed by 4 trails on PU healing,5 on symptom improvement,6 on Hp eradication and 3 on reducing ulcer relapse.Besides,side effects of modified SJZD were reported only in one case.However,due to high risk of bias and limited sample size,the results need to be supported by more rigorous RCTs.(3)The experimental study found that ?compared with those in spleen deficiency gastric ulcer model group,SJZD treatment group rats had smaller gastric ulcer index(p<0.05)and higher quality of ulcer healing with less inflammatory cell infiltration in submucosa of the ulcer base.It indicated that SJZD could promote the healing of stomach ulcer and enhance the ulcer healing quality.?HGF expression in gastric mucosa in rats with spleen-deficiency gastric ulcer was significantly higher than that in the control group(p<0.05),while the content of c-Met was significantly lower(p<0.05).And both HGF and c-Met levels in the SJZD group were significantly higher than those in the model group(p<0.05),which suggested that SJZD could promote HGF maturation,followed by initiation of its combination with c-Met to promote the proliferation and migration of gastric mucosa epithelial cells at the margin of ulcer,thereby accelerating the healing of gastric ulcer.?PI3K expression was higher and p-Akt were significantly higher(p<0.05)at the margin of gastric ulcer in model rats compared with those in normal group.In the SJZD group,the content of PI3K significantly decreased(p<0.05)to the level close to the normal group and p-Akt also decreased.It suggested that SJZD may promote the healing process of ulcer through activating the PI3K/Akt signaling pathway.?The results showed that EGFR level in both ventral hypothalamus nucleus(vhn)and hippocampus CA1 region in model group was significantly higher than that in normal group(p<0.05).While that in the SJZD group was significantly lower than the model group(P<0.05).It suggested that SJZD could correct the high expression of EGFR in vhn and hippocampal CAI region of model rat.? The expression of SSTR2 in vhn and the hippocampus CA1 in the model group significantly decreased compared with that in the control group(p<0.05).While SSTR2 in vhn of SJZD group significantly increased(p<0.05)and SSTR2 in hippocampal CA1 region was also higher than that in the model group.It showed that the SJZD could improve the expression of SSTR2 in the brain of rat with spleen deficiency and gastric ulcer.?In the model group,the expression of TFF3 in hippocampus CA1 region decreased significantly compared with that in the control group(p<0.05).That in SJZD group increased slightly compared with that in model group.While the TFF3 level in vhn of model group did not change significantly compared with that in control group.That in the SJZD also did not vary with difference of statistical significance.It demonstrated that SJZD did not have obvious regulation on TFF3 in the brain of rat with spleen deficiency and gastric ulcer.?And in the model group,the ERK1/2 level of hippocampus CA1 region was significantly lower than that in normal group(p<0.05),and in SJZD group it was even lower with significant difference compared with that in the normal group(p<0.05).In the model group,level of ERK1/2 in vhn increased significantly(p<0.05)compared with that in the control group,while the ERK1/2 expression of vhn decreased significantly(p<0.05)after SJZD treatment.It indicated that SJZD could regulate the overexpression of ERK1/2 in vhn.These results suggested that SJZD could promote the healing of gastric ulcer by modulating EGFR,SSTR,TFF3 and ERK signaling pathway in the central system,thus maintaining the integrity of gastrointestinal mucosa and promoting the repair of gastrointestinal mucosa.Conclusion:(1)The main connotation of "spleen governs protection" is that the spleen can protect the body from pathogens,expel pathogens after catching the disease,promote the disease healing and prevent disease transferring to other zang and fu organs and meridians and collaterals and prevent disease relapse.All the functions of spleen originate from its function of "governing transportation and transformation" and thereby being "the source of qi and blood"to the nourish zang and fu organs,limbs and orifices(2)Systematic review and meta-analyses of RCTs on SJZD confirms that modified SJZD can promote the healing of gastric ulcer,alleviate clinical symptoms,eradicate Hp and reduce ulcer relapse.And modified SJZD is relative safe.But it still needs more rigorous randomized controlled trails to further support the conclusion.(3)The experimental study shows that SJZD promote gastric ulcer healing through facilitating the expression of HGF and c-Met on the edge of ulcer and accelerating the proliferation and migration of gastric epithelial cells.PI3K/Akt signaling pathway are also involved in the process of SJZD promoting gastric ulcer healing.(4)SJZD also promotes the ulcer healing process through regulation of brain intestinal peptide,related signal factors and signal pathway,and thereby promoting the repair of gastrointestinal mucosa damage.It suggests that the biological mechanism of "spleen governs protection " is related with the gut-brain axis and peptide apart from immune system and local gastric mucosal protection.