Research On Protective Functional Mechanism For Mucosal Blenn Barrier Of Bile Reflux Gastritis Rats With Weiyanyin

Objective: this research intend to verify the curative effect that Weiyanyin cure bile reflux gastritis by examining the changes of layer thickness which is masc for AB/PAS dyeing and hexosamine, phospholipid, PGE₂ in stomach mucosa and 6-keto-PGF_1α, TXB₂ in blood plasma. discussing the protective functional mechanism of Weiyanyin on gastric mucosa of bile reflux gastritis ratsMethodsPart 1: The effect of Weiyanyin on masc layer thickness for AB/PAS dyeing and Pathological morphology of gastric mucosa of bile reflux gastritis rats1 HE dyeing40 healthy wistar rats,half male and half female,were randomly divided into 5 groups, 8 rats every group, namely normal group,model group(equal volume physiological saline were given through the means of watering stomach) ,high-dose group of Weiyanyin ( called high-dose group, 21.4g/Kg avoirdupois Weiyanyin drank through the means of watering stomach), low-dose group of Weiyanyin (called low-dose group,10.7g/Kg avoirdupois Weiyanyin drank through the means of watering stomach) , Domperidone control group (called control group, 2.78mg/Kg avoirdupois). Normal group were given equal volume physiological saline through the means of watering stomach , once everyday, do that 35 days .Except for normal group,each group were given reflux liquid 15ml/kg avoirdupois once a day,do that 35 days. The last time, give rats reflux liquid then stop giving them anything for 24 hours. We cut the belly open to obtain stomach by aseptic technique, then poaching Gastric contents with ice- physiological saline, at last, scissors a piece of tissue of sinus ventriculi which is about 1mm3 ,4 percent paraform fixation, dyeing by HE to observe those pathohistology changes of gastric mucosa with light microscope and analysis inflammatory cell of every group.2 AB/PAS dyeing:The dividing groups and medication use the same way as before , The last time,we give rats reflux liquid then stop giving them anything for 24 hours and put them to death by decapitation. We cut the belly open to obtain stomach by aseptic technique,then poaching Gastric contents with ice- physiological saline, and blot those gastric mucosa with filter paper. scissors a piece of tissue of sinus ventriculi which is about 1mm³ , fix these tissue with 4 percent paraform, dye them by AB/PAS dyeing to observe them and measure the masculine layer thickness, in every sheet ,we measure 10 times in different parts and any interval is the same then obtain average value. Part 2: The effect of Weiyanyin on phospholipid and hexosamine in gastric slime layer of bile reflux gastritis ratsThe dividing groups and medication use the same way as before , The last time,we give rats reflux liquid then stop giving them anything for 24 hours and put them to death by decapitation. We cut the belly open to obtain stomach by aseptic technique,then poaching gastric contents with ice- physiological saline, and blot those gastric mucosa with filter paper. Take a quarter of stomach and scrape gastric mucosa with microscope slide, weigh them then add methanol/CHCl3 into a half of them, mix them. take underlayer liquid 1 ml into alimentary tube to measure. Another 1/2 for measuring aminohexose, the thing which is scraped from gastric mucosa increase of 6 mol/L HCl boiling water bath for 1 h, after cooling plus 4 mol/L of sodium hydroxide and to pH 7.0, 3000r/min centrifugal 10 min, take the supernatant 1 ml of analyte.Part 3: The effect of Weiyanyin on contents of PGE2 in gastric mucosa and 6-Keto-PGF_1α, TXB₂ in blood plasm:The dividing groups and medication use the same way as before , The last time,we give rats reflux liquid then stop giving them anything for 24 hours and put them to death by decapitation. We cut the belly open to obtain stomach by aseptic technique, then poaching gastric contents with ice- physiological saline, and blot that gastric mucosa with filter paper. Mucosal were scraped up by reamer and weighted, and 10% issue plasma was preparation in ice bath, which centrifugated immediately for taking serum (3000 rs/ min, 15 mins) and keep temperature of under -20℃to examine PGE₂.After fast for 12 hours, kill it by decapitation and take blood about 5ml, misce bene. Infund it into plastics tube,then which centrifugated immediately for taking serum (3000 rs/ min, 15 mins), separated plasma and blood corpuscle, infund plasma into tube, and keep temperature of under -20℃to examine 6-Keto-PGF_1α, TXB₂ .ResultsPart 1: The effect of Weiyanyin on masc layer thickness for AB/PAS dyeing and Pathological morphology of gastric mucosa of bile reflux gastritis rats:1 HE dyeingMucosal epidermis of rats in normal group was smooth, and without breakage, amotic; Few inflammation cells could be seen; mucosal epidermis of rats in model group was sparseness, attenuation. epidermis cell appeared putrescence, desquamate, and a lot of inflammation cell and atypical hyperplasia could be seen; compared with model group, epidermis cell in mucosal of Weiyanyin low-dose group and Domperidone group was more abundance, and a few inflammation cell could be seen. The mucosal of Weiyanyin high-dose group is similar to normal group. (Fig. 1-5)Semi-Quantitative Analysis showed that, compared with normal group, inflammatory cell infiltration in mucosa of model group was much more obviously (P<0.01); compared with model group, inflammatory cell infiltration of Weiyanyin high-dose group, low-dose group and Domperidone control group was less (P<0.01 or P<0.05); And inflammatory cell infiltration of Weiyanyin high-dose group was less than Domperidone control group (P<0.05). (Table1)2 AB/PAS dyeingMasc layer thickness for AB/PAS dyeing of rats in model group is thin, even absent. The table 2 showed,compared with model group, Masc layer thickness for AB/PAS dyeing of rats in Weiyanyin high-dose group,low-dose group,Domperidone is better (P<0.01 or P<0.05),and Weiyanyin high-dose group is the best (P<0.01).Part 2: The effect of Weiyanyin on phospholipid and hexosamine in gastric slime layer of bile reflux gastritis rats1 The comparison of the content of phospholipid in gastric slime layer of bile reflux gastritis ratsThe table 3 showed, compared with normal group, the content of phospholipid of model group was lower, which had a very significant difference (P<0.01). Compared with model group, the content of phospholipid in high-dose group was higher, which had a very significant difference (P<0.01), Domperidone group and low-dose group also was higher, which had a significant difference (P<0.05).There had no marked difference (P>0.05) of the content of phospholipid among low-dose group and Domperidone group.2 The comparison of the content of hexosamine in gastric slime layer of bile reflux gastritis ratsThe table 4 showed, compared with normal group, the content of hexosamine of model group was lower, which had a very significant difference (P<0.01). Compared with model group, the content of hexosamine in high-dose group was higher, which had a very significant difference (P<0.01), Domperidone group and low-dose group also was higher, which had a significant difference (P<0.05).There had marked difference of the content of hexosamine among low-dose group and Domperidone group (P<0.05).Part 3: The effect of Weiyanyin contents of PGE2 in gastric mucosa and 6-Keto-PGF_1α, TXB₂ in blood plasm:1 The comparison of the content of PGE₂ in mucosal of bile reflux gastritis ratsThe table 5 showed, compared with normal group, the content of PGE₂ of model group was lower, which had a very significant difference (P<0.01). Compared with model group, the content of PGE₂ in high-dose group,low-dose group and Domperidone group was higher, which had a very significant difference (P<0.01).There had a very significant difference of the content of PGE₂ among low-dose group, high-dose group and Domperidone group(P<0.01).2 The comparison of the content of TXB₂ , 6-Keto-PGF_1αin blood plasm of bile reflux gastritis ratsThe table 6 showed, Compared with model group, the content of 6-Keto-PGF1αin high-dose group,low-dose group and Domperidone group was higher, which had a very significant difference (P<0.01 or P<0.05 ). The content of TXB₂ in high-dose group,low-dose group and Domperidone group was lower, which had a very significant difference (P<0.01 or P<0.05 ).There had a very significant difference (P<0.01)of the content of 6-Keto-PGF_1αand TXB)2 among high-dose group and Domperidone group.Conclusion:1 Weiyanyin has a good curative effect of treating bile reflux gastritis,and it could improve masculine layer thickness for AB/PAS dyeing of rats, and relieve the infiltration of phlegmasia cell,and improve pathomorphism of gastric mucosa and protect mucosa of experiment bile reflux gastritis rats effectively.2 Weiyanyin could increase the content of hexosamine and phosphatide, so that strengthen the glutinousness and hydrophobicity of stomach mucago, and protect mucosa of experiment bile reflux gastritis rats effectively.3 Weiyanyin could increase the content of PGE₂ and 6-Keto-PGF_1α,cut down the content of TXB₂ and protect mucosal of experiment bile reflux gastritis rats effectively.