Investigation Of JianWeiYuYang Granule's Effects On Peptic Ulcer Recurrence And The Relative Study On Hydrophobicity Of Gastric Mucosa

Objective:To investigate the effect of Jianweiyuyang(JWYY) granule on the gastric ulcer recurrence rats and the peptic ulcer patients and the expression of PAF and PS2 in the ulcer healing process and study the effect of JWYY granule on the contents of aninohexose,phosphatide and Contact angle(CA),in order to reveal the mechanisms of JWYY's effects on ulcer recurrence from the view point of hydrophobicity.Methods:1.Animal Experiment:108 rats were randomly divided into five groups:control group (n=12),sham operated group(n=12),model group(n=36),Ranitidine group(n=24) and JWYY group(n=24).All rats were induced ulcer by the method of Okabe acetic squash method with some modification except for control group and sham operated group.Rats of JWYY and Ranitidine group were intragastric administrated JWYY physic liquor(1.62g/(kg.d)) and Ranitidine physic liquor(27mg/(kg.d)) respectively,one time per day,totally 90 days.The others were intragastric administrated distilled water.At 91^st day,rats of model group were randomly divided into model group and model recurrence group.Gastric ulcer recurrence model was duplicated by IL-1β(1μg/kg) intraperitoneal injection except for rats of model group which were intraperitoneal injection saline instead,At 7^th,92 ^th day(92^th day is also 48h later after ulcer recurrence was induced),the rats of each group were put to death and the stomach was taken out.After get the liquid of upper gastric mucosa,then stomach wall of rats were devided into three parts.A part of stomach wall was put into the 4% formaldehydum polymerisatum for 16 hour,then it was embedded into the paraffin wax for HE staining and measure the expression of PAF and PS2 by immunohischemical.Then a part of stomach wall was put into the liquid nitrogen kettle for measure the expression of PAF and PS2 by RT-PCR and Western blotting method.The last part of stomach wall was used to measure the contact angle of gastric mucosa and the contents of aninohexose and phosphatide in ulcerated gastric mucosa for examing the hydrophobicity of gastric mucosa.2.Clinical Research:Choose 76 gastric ulcer patients(final diagnosis by gastroscope) were randomly divided into two groups,including JWYY treatment group,(36 cases,treatment group) and western drug control group(36 samples,control group).12 patients without gastric mucosa lesion was named normal group.Including 50 Hp-positive PU patients were confirmed by endoscopy,histological inspection,urease test,the differentiation of symptoms and signs of the traditional Chinese medicine is the stagnation of liver-QI with deficiency of the spleen type.JWYY was used for treatment group,9g a time and 3 times a day and Ranitidine used for control group,0.15g a time and twice a day.All medicine used four weeks as a period of treatment and Amoxicillin and Metronidazole were used for the patients with lip infection additionally in Rani group.We had investigated the rate of ulcer recurrence,curative effect under gastroscope,eradicative rate of Hp,clinical curative effect in Chinese medicine syndrome.We prepared four or five pieces of tissues.They were cut from the part of gastric ulcer before treatment from two groups,one piece was used in Hp urea enzyme experiment,one pieces were fixed with 4%paraformaldehyde sotution,then wrapped up with paraffin wax and sectioned for immunohischemical,to observe the change of PAF,Ps2.one piece was used in measuring the contact angle of gastric mucosa,and the contents of aninohexose and phosphatide in ulcerated gastric mucosa for examining the hydrophobicity of gastric mucosa,another one or two pieces were stored at liquid nitrogen kettle for RT-PCR and Western blotting.To cut four or five pieces of tissues from the part of gastric organ before treatment as normal group,which were dealed with the same method and they did not treat or recheck in the whole processes.The same processes have done after a period of treatment.Results:1.Experiments study:After 7 days when the recurrence model was caused,the other groups can obeserve the ulcer foci of gastric mucosa in model rats except control group and sham operated group,the ulcer foci in rats were observed macroscopically and histologically at different degree.GUI of model group is the highest,while the GUI of JWYY group and Ranitidine group were both significantly lower than that of model group(P＜0.01) Histo-morphology of both JWYY group and Ranitidine group were better than that of model group(P＜0.01).At the 92^th day,ulcer relapse rate was 75%,16.7%,58.3%,in JWYY group,Model recurrence group and Ranitidine group respectively,there was significant difference between the Model recurrence group and the two treatment groups(P＜0.01).While the structure of JWYY group was relatively regulation compared with Ranitidine group The rate of the recurrence of ulcer and the ulcer index were decreased significantly in JWYY group compared with Ranitidine groups(P＜0.01).The contents of aninohexose,phosphatide and Contact angle(CA) increased in JWYY group,there was significant difference between JWYY group and Ranitidine group(P＜0.01).immuno0-hischemical, RT-PCR and Western blotting method show that There has a Linear correlation between the the contents of phosphatide, aninohexose,Contact angle(CA) and the expression of PAF,PS2 mRNA in rats(P＜0.01).JWYY granule can prevent the occurrence and relaps of ulcer and affect the hydrophobicity of gastic mucosa and strengthen the stability of myxo-gellayer by reducing the express of PAFmRNA,elevating the express of PS2mRNA and effecting the contents of phosphatide and aninohexose and Contact angle(CA),this is maybe one of the mechanisms of JWYY to heal ulcer quickly.2.Clinical research: Through the clinical investigation we can known that both treatment group and control group have fine effect on gastric ulcer and there are no significant difference in the rate ofventriculi curative effect(P＞0.05),but treatment group is better in ameliorate the clinical symptom than the control group,furthermore the compliance of the patients is better in JWYY group than control group(P＜0.01).The rate of the recurrence is 4.1%in treatment group and 30%in control group(P＜0.01). furthermore optical microscope investigation show that there is litter inflame- matory cell infiltration and the structure of PU patients was relatively regulation compared with other groups in JWYY group.The contents of aninohexose,phosphatide and Contact angle(CA) of PU patients increased in JWYY group,there was significant difference between JWYY group and Ranitidine group(P＜0.01).The immuno-hischemical, RT-PCR and Western blotting method show that There has a Linear correlation between the the contents of phosphatide,aninohexose, Contact angle(CA) and the expression of PAF,PS2 mRNA in PU patients(P＜0.01).JWYY granule can prevent the occurrence and relaps of ulcer and affect the hydrophobicity of gastic mucosa and strengthen the stability of myxo-gellayer by reducing the express of PAFmRNA,elevating the express of PS2mRNA and effecting the contents of phosphatide and aninohexose and Contact angle(CA),this is maybe one of the mechanisms of JWYY to heal ulcer quickly.Conclusions:1.JWYY granules can accelerate the ulcer healing,ameliorate the structure of new mucosal organize and prevent the recurrence of gastric ulcer through animal experiment and clinic observation.2.JWYY granule can prevent the occurrence and relaps of ulcer and affect the hydrophobicity of gastic mucosa and strengthen the stability of myxo-gellayer by reducing the express of PAF,elevating the express of PS2 and effecting the contents of phosphatide and aninohexose and Contact angle(CA),this is maybe one of the mecha- nisms of JWYY to heal ulcer quickly.