| ObjectiveChronic atrophic gastritis(CAG)is a key stage in the evolution of chronic gastritis to gastric cancer,so blocking and reversing the progression of CAG is an effective way to prevent the occurrence of gastric cancer.Now there is no systematic and effective treatment for the disease in western medicine,while traditional Chinese medicine(TCM)has a significant advantage in this disease.There is a strong association between the genesis and development of CAG and gastric acid secretion.Our team previously found that Ezrin-ACAP4 pathway played an important role in gastric acid secretion.So it is of great significance to study the mechanism of TCM treating CAG based on Ezrin-ACAP4 pathway.Firstly,the core pathogenesis and therapeutic method of CAG was obtained by identifying the core location and nature of disease of CAG according to the clinical study of CAG’syndrome elements.Secondly,based on the core pathogenesis and therapeutic method of the clinical findings,the effective prescription of CAG-Xiaopiling(Lanceolata,Lily,Salyia,Hedyotis diffusa willd,Citrus medica,Combined spicebush,Panax notoginseng powder,Barbed skullcap,Zedoary turmeric)was chosen and the effect of Xiaopiling granule was observed during the progression of CAG in rats.Finally,the level of gastric acid secretion was investigated after the administration of Xiaopiling granule in CAG rats and the regulation mechanism of Ezrin-ACAP4 pathway related to gastric acid secretion was explored in vivo.This research would not only provide guidance for clinical diagnosis and treatment of CAG,but also provide objective evidence for clinical application of traditional Chinese medicine to regulate gastric acid secretion in order to prevent and treat CAG,so as to guide the clinical use of TCM in CAG.Methods1.Clinical Research:A multi-center and large-sample cross-sectional survey was applied to collect the four diagnostic information of patients with chronic atrophic gastritis and to establish a database for four diagnostic information.Factor analysis was used to analysis the four diagnostic of the patients with CAG,and thereby obtaining syndrome elements,identifying the core location and nature of disease and getting the core pathogenesis and therapeutic method.2.Experimental Research:①38 male SPF Wistar rats were randomly divided into two groups,including control group and model group;both of them have 19 rats.MNNG comprehensive method was used to establish CAG model in model rats.The change of characterization,body weight,food intake,water intake and grip strength were recorded every week.Gastric histopathology was observed in 5 randomly rats of both groups in the end of 16 week to clarify the CAG rat model successfully,and to judge the syndrome.②120 male SPF Wistar rats were randomly divided into two groups,with 25 rats in control group and 95 rats in model group.MNNG comprehensive method was used to establish CAG model in model rats.The CAG rat model was clarify successfully in the end of the 16th week.6 control rats and 24 model rats were selected to complete the experiment in the CAG phase(Other model rats continue to take MNNG).24 model rats were randomly divided into three groups,including Xiaopiling prevention-treatment group(PT-group),Weifuchun PT-group and Blank model group(PT-group),each have 8 rats.Three groups were administered with 4.3ml/kg Xiaopiling solution,4ml/kg Weifuchun solution and 4ml/kg water by gavage,qd.The rats were put to death for sampling after 8 weeks administering.Other model rats were clarified in the PLGC phase in the end of the 26th weeks,which were randomly divided into six groups,including Xiaopiling PT-group,weifuchun PT-group,Blank model group(PT-group),Xiaopiling treatment group,weifuchun treatment group,natural recovery group,each group have 9 rats and was administered with above drugs and doses For eight weeks.All rats were drawn materials in the end of 34th week.③To observe the pathological features of gastric tissue of rats in each group after the administration of Xiaopiling in the different stages of CAG.The level of gastric acid was measured with a pH instrument,the expression of H,K-ATPase,Ezrin,ACAP4 and ARF6 in Rat gastric mucosa was detected with Immunohistochemistry and Western-blotting.Results1.Clinical Research:In cross-sectional study,a total of 413 eligible CAG patients were collected.The syndrome elements of location of disease with CAG patients were in sequence of "stomach>spleen>liver>heart>colorectal>intestinal>kidney".The syndrome elements of nature of disease with CAG patients were in sequence of "qi deficiency>qi stagnation>yin deficiency>heat>dampness>blood stasis".2.Experimental Research:①In the early stage of model(the 3th-9th weeks),model rats began to form thin,decrease of grasping strength,reduction of food intake,reduced mobility,pale tongue with white,pale lips,yellow and reluster hair,pale claws which continued until the end of model.It was speculated that syndrome characteristics of model rats were Qi-deficiency in the early stage of model;In the later stage of model(the 13 th-15th weeks),model rats appeared blue lips,dull-red tongue,blue claws.It was speculated that model rats appeared blood stasis on the basis of the syndrome of Qi-deficiency in the later stage of model.Therefore,the syndrome of the CAG rats which’s molded by MNNG comprehensive method were Qi-deficiency and blood stasis syndrome.②The rats treated by Xiaopiling had gotten fatter,more shine hair,more sensitive,and more increased activity,pinker lips and claws,rosier tongue,steadier growth in weight,more food intake than before.Body weight and food intake increased higher than blank model group,natural recovery groups.Obviously,Xiaopiling could significantly improve the general status of the CAG and PLGC rats.③Rat gastric histopathology results:After the treatment of CAG stage rats,3 rats in Xiaopiling PT-group had no significant anomalies in rat gastric mucosa(3/5,60%).2 rats were diagnosed with CAG(2/5,40%);5 rats in Weifuchun PT-group were diagnosed with CAG(5/8,63%).2 rats were diagnosed with PLGC(2/8,25%).1 rat had no significant anomalies in rat gastric mucosa(1/8,12%);6 rats in blank model group were diagnosed with dysplasia of different degrees(6/7,86%).1 rat was diagnosed with CAG(1/7,14%).After the treatment of PLGC stage rats,4 rats in Tiaoqi Huoxue prescription PT-group appeared mild to moderate dysplasia(4/7,56%),3 rats were diagnosed with CAG(3/7,44%);1 rats in Weifuchun PT-group appeared gastric cancer(1/7,14%).6 rats appeared mild to moderate dysplasia(6/7,86%);3 rats in blank model group were diagnosed with gastric cancer(3/8,38%).5 rats were diagnosed with dysplasia of different degrees(5/8,62%);7 rats in Xiaopiling treatment group were diagnosed with CAG(7/9,78%).2 rats were diagnosed with mild dysplasia(2/9,22%);5 rats in Weifuchun treatment group appeared mild to moderate dysplasia(5/9,55%).4 rats were diagnosed with CAG(4/9,45%);7 rats in natural recovery group were dysplasia of different degrees(7/7,100%).Obviously,Xiaopiling could stop and reverse the progression and pathological state of the CAG rats.④The gastric acid pH of the rats treated by Xiaopiling valued significantly higer than blank model group,natural recovery group and Weifuchun group(P<0.05).Obviously,Xiaopiling could significantly improve the levels of gastric acid secretion in CAG and PLGC rats.⑤The levels of H,K-ATPase expression in gastric tissue of rats which treated by Xiaopiling were higher than blank model group,natural recovery group and Weifuchun group(P<0.05);The levels of earin expression in gastric tissue of rats which treated by Xiaopiling were higher than blank model group,natural recovery group and Weifuchun group(P<0.05);The levels of ACAP4 expression in gastric tissue of rats which treated by Xiaopiling were higher than blank model group,natural recovery group and Weifuchun group(P<0.05);The levels of ARF6 expression in gastric tissue of rats which treated by Xiaopiling were higher than blank model group,natural recovery group(P<0.05).Obviously,Xiaopiling could significantly enhance H,K-ATPase,Ezrin,ACAP4,ARF6 protein expression levels in gastric mucosal of CAG and the PLGC rats.Conclusion①There were deficiency and excess syndrome in the pathogenesis of CAG.Deficiency syndrome included qi deficiency and yin deficiency.Excess syndrome included qi stagnation,heat,dampness and blood stasis.We should diagnose and treat CAG based on "differentiation of syndromes to find out the cause of disease,differentiation of superficiality and origin,reinforcing the healthy qi to restore the normal function,eliminating pathogenic factors to treat symptoms,intermingled tonifying and obstruction-removing therapy".②The syndrome characteristics of CAG rat model established by MNNG comprehensive method was Qi deficiency and blood stasis.Xiaopiling granule could obviously improve the general condition of CAG and PLGC rats,increase the level of gastric acid secretion in rats,prevent and reverse the disease progression and pathological state of CAG rats,and increase the expression level of Ezrin in gastric mucosa of rats.Ezrin could recruit more ACAP4 positioning in the apical membrane of gastric parietal cells through the phosphorylation of the 66th Ser.ACAP4 could regulate the vesicle transport function of ARF6 in gastric parietal cells through a GTP hydrolysis,so that more sac vesicles were transported from the cytoplasm to membrane,then H,K-ATPase in the vesicles anchored in the apical membrane,so as to enhance the gastric acid secretion,which could delay the progression of CAG,and prevent the occurrence of gastric cancer. |
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