The Experimental Study On WeiWeiFang Decoction On The Contents Of Hexosamine In Ulcerated Gastric Mucosa

Research backgroundPeptic ulcer is a clinically common chronic relapsing disease. In recent years, gastric mucosal protective agents, H2 receptor antagonist (H2R) and proton pump inhibitor (PPI) applications and standards to enable the anti-HP treatment significantly improve short-term effect, but it is still difficult to avoid ulcer recurrence, drug withdrawal 1 year later, the recurrence rate reach up to 30% ~ 50%, besides the drug costs and side effects, poor patient compliance problems exist, so the disease recurrence rate remains high, so look for the treatment of PU and the development of natural drugs become urgent and important task.The pathogenesis of peptic ulcer is complex. At present, balance theory of intragastric attack factors and defense mechanisms is being widely accepted. Hydrophobicity of gastric mucosa has an important value of the defense, in the incidence of ulcer development and recurrence of the process plays an important role. Strong hydrophobicity of gastric mucosa, the gastric mucosal defensive ability, ulcers is easyly to be natural repaired, recurrence is not easy,on the other hand, are easily lead to ulcers, also it is difficult to cure, even if short-term cure, but also easy to relapse. Gastric hydrophobic basic substances are lipids from mucus gel material (the main components are phospholipids) and glycoprotein (the main components are hexosamine). Previously research proved: HS content and ulcer index was a negative correlation, and reflects the integrity of the gastric mucosal membrane potential was a positively correlation, HS content has become widely used to determine gastric mucosal injury and one important parameters of Pharmacodynamic study.WWF is a empirical prescription, which is used to cure peptic ulcer in clinic by my tutor. It is made up of Fuling,Pugongying,Baiji,Sanqi. The study aims to investigate the effect of WWF on HS in experimental gastric ulcer on rats and probe the possible mechanism of WWF repair stomach mucosa damage. The rats are induced by icy acetic acid, HS content of gastric mucosa by colorimetric determination, Meantime observe the general situation of GU healing, Secretion of gastric acid is detected by PH paper, analysis the changes of gastric mucosa pathological tissue slice, aims to clarify the clinical efficacy. Adopt different dosage aims to explore the defense mechanism of the gastric mucosa intervention role, Provide a theoretical basis for clinical application in the further.Methods1. Animal grouping and administration Fifty-six male Wistar rats with SPF level and at the weight of 180-220g are divided into 7 groups according to a completely random method. Those are normal group, model group, self-healing group, WWF group with high, medium and low dose and ranitidine group. Except normal group , rats of other groups were induced by icy acetic acid. In each group, there are 8 rats. After the surgery,rats can eat freely ,while those of WWF groups and ranitidine group were administrated various doses of WWF and ranitidine suspension(1ml per 100g body weight) for a week (once everyday) from the forth day. Except model group were executed in the forth day, the others were at the eleventh day.2. Preparation of GU model Models were prepared according to the classic glacial acetic acid method, and transformed. The rats must be fasted diet over 24 hours before the operatation. Inject intraperitoneally 10% of Chloral Hydrate to anaesthetize rats, preparefor abdominal skin and disinfect with alcohol ,then cut one 2cm longitudinal incision since xiphoid along ventral, expose the stomach, and paste acetic acid-soaked filter in diameter of 5 mm in the tunica serosa gastria, 1minter later, put the stomach back, suture layer by layer, disinfect the suture department again.3. Indicators detection Removed stomach after the rats were killed, Secretion of gastric acid was detected by PH paper. Their stomach were fixed with formalin of 10 percent, embedded with olefin and sliced up. The tissue indexes were tested, pathology examination by HE dyeing. Gastric mucosa and mucus gently scraped with slide, weighing them under low temperature, placed in 2ml normal saline, prepare homogenate by routine, then put it into glass test tube, 3000r/min centrifugal 10min, the supernatant fluid reserve at -20℃, respectively in the determination of protein and HS content.Statistical analysisThe experimental results are expressed by X|-±S. SPSS13.0 software was applied into One-way ANOVA for mean comparison based on multiple samples. When the variance was homogeneous, Least-significant difference (LSD) was applied for test and the value of P≤0.05 would mean significant difference.Results1. The changes of PH level The study demonstrated that gastric fluid PH value of Model group increased significantly higher than the normal group(P<0.01). Weiweifang with middle dose has no discrepancy compared with normal group (P> 0.05) . Ranitidine group compared with the model group has significant difference (P <0.05). Weiweifang with high-dose group and the ranitidine group, no significant difference (P> 0.05).2. Observation of gastric mucosa Gastric mucosa of normal control group rats were normal and smooth, without congestion, edema and erosion. Gastric mucosa of model group rats were congested and dropsical with many bleeding dots and ulcers; some material of light coffee color could be observed inside stomach. Self-healing group demonstrate poor healing, could see clearly ulcer department, were congested with many bleeding dots and ulcers; observed inside stomach. gastric mucosa organization adhesion obviously. In groups of WWF with high and middle dose, rats' gastric mucosa were smooth with little gore and bleeding dots. In groups of WWF with low dose, rats' gastric mucosa were congested and dropsical lightly, with little gore and bleeding dots. In Ranitidine group, rats' gastric mucosa were smooth without obvious erosions.3. Observation under microscope Gastric mucosa of normal group rats were normal and smooth,with gland in order. Inflammatory cells and bleeding inclination could not be observed. Gastric mucosa of model group rats were not intact, with many red cells on the surface of mucosa. The glands were broken through, and capillary vessel under ulcers were blocked and bleeding. Blots might be observed inside some little aorta. Some ulcers could destroy muscle layer, the muscle cells lined in disorder. Self-healing group rats' gastric mucosa was broken through, with gland cells were arranged in disorder and inflammatory cells infiltration. Some of fibrous scar formation and occasionally thrombosis is observed. In groups of WWF with high and middle dose, we found that epidermic cell of gastric mucosa were tumid and the clearance between glands were enlarged. Other tissue and the micro-circulation under mucosa were normal. In groups of WWF with low dose, under-mucosa layer of rats' gastric mucosa were destroyed with light pathology changes. Ranitidine group has no special discrepancy compared with group WWF with high dose. Gastric mucosa were smooth, with gland in order.4. Hexosamine(HS) content of gastric mucosa Compared with model group, WeiWeiFang (WWF) can be significant increase in gastric mucosal HS content (P <0.05), obviouly in the group of WWF with middle dose (P <0.01); group of WWF with high-and low-dose group and the ranitidine group had no statistical significance (P> 0.05); self-healing group compared with each treatment group significantly different (P <0.05), and with the middle dose group the most significant difference (P<0.01).Conclusion1. The method of replicating gastric ulcer model by icy acetic acid was in line with performance of human gastric ulcer, which has a high stability and good feasibility.2. Model group is higher in gastric fluid pH than normal group. It manifests that acetic Acid-Induced Gastric ulcer is a stress for rats. For self-protect, rats irritably remove the equilibrium mechanism to counteract Acetic Acid. It may be the reason why the model group is in higher lever in gastric fluid pH.3. General observation on gastric mucosa shows that ulcer healing is in good condition in every medication group. WWF mid-dose group is the best which is followed by WWF high-dose group and the ranitidine group, and there is an obvious distinction between medication groups and self-healing group.4. The group of WWF with middle dose has the best therapeutical effect by detecting gastric ulcer circa histopathologic slide in light microscope. It has good effect in microcirculation, anti-inflammatory and cell regeneration.5.Each group of WWF, gastric mucosa HS content was significantly higher than the model group, also higher than the ranitidine group, indicate that WWF may promote HS content increased, and its mechanism may be related to maintaining the hydrophobicity of gastric mucosa and gastric mucosal integrity, the effective better than western medicine.In above, it has been find out that The group of WWF with middle dose has the best effect by detecting gastric mucosa, gastric ulcer circa histopathologic slide. WWF have shown that a good organization of the ulcer repair, the HS contents of gastric mucosa were significantly higher than the model group, show that the drug can increas HS content, there is the role of gastric mucosal protection. Especially, WWF with middle dose group has significant effect, compared with the western medicine group, gastric mucosal HS were significantly higher indicating more effective than the western medicine group (ranitidine group). This experiment proof WWF have anti-ulcer action may through enhance gastric mucosal defensive factors.