The Effect Of Weiyanyin On Gastrointestinal Peptide Of Bile Reflux Gastritis Rats

Objective: Weiyanyin is composed of bupleurum chinense, nutgrass galingale rhizome, fructus meliae toosendan, corydalis tuber, atractylodes macrocephala, hoelen, prepared radlx glycyrrhizae, rhizoma coptidis, aurantii nobilis pericarpium, aurantii fructus, prepared rhizoma pinelliae with ginger, stir-baked caulis banbusae in teaniam with ginger juice, et al. In traditional Chinese medicine, it has the effects to disperse the depressed liver-energy, cholagogue, invigorate the spleen, tonifying didney pneuma, harmony stomach, and calm the adverse-rising energy. It is an effective prescription sieved by professor after many years clinical practice, which has good therapeutic efficacy on treating bile reflux gastritis. This research applies reflux liquid to duplicate the model of bile reflux gastritis rats by intragastric administration. The aim is to verify the curative effect of Weiyanyin by observing the changes of pathological morphology of gastric mucosa, motilin (MTL) in blood plasma, substance P (SP) in gastric mucosa, prostaglandin E2 (PGE2) in gastric mucosa and gastrin (GAS) in blood serum of bile reflux gastritis rats. And then discuss the functional mechanism of Weiyanyin curing bile reflux gastritis from the perspective on adjustment of gastrointestinal peptide. MethodsPart 1: The effect of Weiyanyin on pathological morphology of gastric mucosa of bile reflux gastritis rats40 healthy wistar rats,half male and half female, were randomly divided into 5 groups, 8 rats every group, named normal group, model group, high-dose group (21.4g/Kg avoirdupois Weiyanyin intragastric administration), low-dose group (10.7g/Kg avoirdupois Weiyanyin intragastric administra- tion), control group (2.78mg/Kg avoirdupois Domperidone intragastric administration). Meanwhile normal group and model group were given equal volume physiological saline by intragastric administration from the second week to the fifth week, once a day Sustained 28 days. Except for normal group, each group was given reflux liquid 15ml/kg avoirdupois by intragastric administration once a day, for 35 days. After the last time administration, we stopped giving the rats food and drink for 24 hours. We cut the belly open to obtain stomach by aseptic technique, then sheared the stomach along arcus major ventriculi, rinsed gastric contents with ice-physiological saline, blotted it with filter paper and macroscopic observed it. At last, two pieces of tissues of sinus ventriculi were scissorsed, each piece about 1mm³. The tissues were fixed with 4 percent paraform, dehydrolysised with alcohol, paraffin imbedding, and cut into sheets. And the sheets were dyed by HE and AB/PAS dyeing respectively to observe pathological changes of gastric mucosa with light microscope. Detected inflammatory cells of every group by semiquantitative analysis through HE dyeing and measured the AB/PAS positive layer thickness with micrometer though AB/PAS dyeing. In every sheet, we measured 10 times in different parts and each interval was the same, and then obtained average value.Part 2: The effect of Weiyanyin on MTL in blood plasma and SP in gastric mucosa of bile reflux gastritis ratsThe dividing groups and medication were same to above. After the last time intragastric administration, stopped giving the rats food and drink for 24 hours and put them to death by decapitation and obtained blood 5ml. Added 100g/L EDTA-2Na 0.2ml to the blood and jolted in plastic tube. Centrifuged the blood (3500 r/min, 15min) and conserved the plasma under -20℃to examine the content of MTL later. We cut the belly open to obtain stomach by aseptic technique, sheared the stomach along arcus major ventricul, rinsed gastric contents with ice-physiological saline, and blotted the tissue with filter paper. Scraped mucosal tissues of sinus ventriculi with microscope slide, weighed them then put them into homogenizer, added physiological saline into them, mixed them, made them into 10 percent of homogenate in ice bath. Centrifuged the homogenate (3500 r/min, 15min) then took clear supernatant liquid into cryogenic box to conserve under -20℃for checking the content of SP later.Part 3: The effect of Weiyanyin on PGE2 in gastric mucosa and GAS in blood serum of bile reflux gastritis rats: The dividing groups and medication used the same way as above. After the last time administration, fasted the rats for 24 hours and put them to death by decapitation and obtained blood 5ml. Centrifuged the blood (3500 r/ min, 15 min) to make serum separated and conserved the serum under -20℃to detect the content of GAS. We cut the belly open to obtain stomach by aseptic technique, sheared the stomach along arcus major ventricul, and then rinsed gastric contents with ice-physiological saline, and blotted the gastric mucosa with filter paper. Mucosal tissues of sinus ventriculi were scraped by microscope slide and weighted. And 10% tissue bomogenate was prepared in ice bath, which centrifugated immediately to take supernatant (3500r/min, 15 min) and conserved under -20℃to examine PGE2 later.ResultsPart 1: The effect of Weiyanyin on Pathological morphology of gastric mucosa of bile reflux gastritis rats:1 HE dyeingMucosal epidermis of rats in normal group was smooth, without breakage, amotic and few inflammation cells could be seen. Mucosal epidermis of rats in model group was sparseness, attenuation. Epidermis cells appeared breakage, desquamate, and a lot of inflammation cells could be seen. Compared with model group, epidermis cells in mucosa of high-dose group, low-dose group and control group were better, and less inflammation cells could be seen. The mucosa of high-dose group was 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 high-dose group, low-dose group and control group was less (P<0.01 or P<0.05); And inflammatory cell infiltration of high-dose group was less than control group (P<0.05). (Table 1)2 AB/PAS dyeingAB/PAS positive layer thickness of normal group was thick and consecutive. It was thin even absent in model group. AB/PAS positive layer thickness of high-dose group, low-dose group and control group was thin slightly, and high-dose group was the best among the three groups. (Fig. 6-10)Measurements of AB/PAS positive layer thickness showed: Compared with normal group, AB/PAS positive layer thickness in mucosa of model group was much thinner (P<0.01); The figures of high-dose group, low-dose group and control group were lower than model group (P<0.01 or P<0.05); High-dose group and low-dose group were higher than control group (P<0.01 or P<0.05). (Table 2)Part 2: The effect of Weiyanyin on MTL in blood plasma and SP in gastric mucosa of bile reflux gastritis rats1 The comparison of the contents of MTL in blood plasma among groupsThe table 3 showed: Compared with normal group, the content of MTL in model group was lower, which had statistical significance (P<0.01); Compared with model group, the contents of MTL in high-dose group, low-dose group and control group were higher, which had statistical significance (P<0.01 or P<0.05); Compared with control group, the contents of MTL in high-dose group and low-dose group were higher (P<0.05).2 The comparison of the contents of Substance P in gastric mucosa among groupsThe table 4 showed: Compared with normal group, the content of SP in model group was lower, which had statistical significance (P<0.01); Compared with model group, the contents of SP in high-dose group, low-dose group and control group were higher, which had statistical significance (P<0.01 or P<0.05); Compared with control group, the contents of SP in high-dose group and low-dose group were higher but had no statistical significance (P>0.05).Part 3: The effect of Weiyanyin on PGE₂ in gastric mucosa and GAS in blood serum of bile reflux gastritis rats1 The comparison of the contents of PGE₂ in gastric mucosa among groupsThe table 5 showed: Compared with normal group, the content of PGE₂ in model group was lower, which had statistical significance (P<0.01); Compared with model group, the contents of PGE₂ in high-dose group, low-dose group and control group were higher, which had statistical significance (P<0.01 or P<0.05); The contents of PGE₂ in high-dose group and low-dose group were higher than control group, which had statistical significance (P<0.01 or P<0.05).2 The comparison of the contents of GAS in blood serum among groupsThe table 6 showed: Compared with normal group, the content of GAS in model group was lower, which had statistical significance (P<0.01); The contents of GAS in high-dose group, low-dose group and control group were higher than model group, which had statistical significance (P<0.01 or P<0.05); Compared with control group, the contents of GAS in high-dose group and low-dose group were higher, which had statistical significance (P<0.01 or P<0.05).Conclusion1 Weiyanyin has a certain curative effect. It could improve the changes of pathomorphology in gastric mucosa, relieve the damnification and inflammatory cell infiltration of gastric mucosa, and increase AB/PAS positive layer thickness. It has the function of protecting gastric mucosa of bile reflux gastritis rats effectively.2 Weiyanyin could increase the content of MTL in blood plasma and SP in gastric mucosa of bile reflux gastritis rats. It could enhance dynamia of stomach intestine, accommodate MMC and promote gastric emptying, speed up the clearance of reflux liquid. So it could decrease the attack chance of reflux liquid, relieve the damage of stomach mucosa.3 Weiyanyin could increase the content of PGE₂ in gastric mucosa and GAS in blood serum of bile reflux gastritis rats. It could strengthen barrier of stomach mucosa, improve blood flow, and encourage the recovery of mucosa. It has the function of protecting and nourishing gastric mucosa.