| Objective: To prove the curative effect of Weiyanyin on bile reflux gastritis and further for the curative effect mechanism on gastric mucosa of bile reflux gastritis by observing the effect of Weiyanyin on free radical metabolism and inflammatory cell factors in gastric mucosa of experiment bile reflux gastritis rats.Methods:Part 1: The effect of Weiyanyin on pathological morpholo- gy of experiment bile reflux gastritis rats.40 healthy clean grade Wistar rats,bisexual and each half, were randomly divided into 5 groups,8 rats in every group, namely normal group, model group (equal volume physiological saline was given through the means of watering stomac- h),high-dose group of Weiyanyin (called high-dose group, 21.4g/kg avoirdupois Weiyanyin was given through the means of watering stomach),low-dose group of Weiyanyin (called low-dose group,10.7g/kg avoirdupois Weiyanyin was given through the means of watering stomach), and Domperidone control group (called control group, 2.78mg/kg Domperidone was given through the means of watering stomach). From the second week to the fifth week, each treatment group was given medicine through the means of watering stomach, once a day. Reflux liquid was given to each group through the means of watering stomach except for normal group according to 15ml/kg avoirdupois, once a day, last for 35 days. Each group was fast for 24hs after watering stomach for the last time, then the head of rat was cut off to obtain stomach from abdomen, and the stomach was cut open along stomach big curve, washed by physiological saline, and bloted up by filter papers. At first, mucosal general instance was observed by eyes, then some mucosa was cut up, and fixation in 4% formaldehyde, dyeing by HE. At the last, mucosal pathological histology was observed by microscope. According to the records, 4 pathological photographs of each rat was randomly drew-off, 32 pathological photographs in all. Detected inflammatory cells infiltration by semi-fixed quantity. Inflammatory cells infiltration extent: Light range: less inflammatory cells could be seen and they were limited in membrana mucosa superficial layer, didn't exceed one third of mucous layer; Midrange: more inflammatory cells could be seen and they were in two thirds of mucous layer; Pond range: a lot of inflammatory cells could be seen and they were in all of mucous layer.Part 2: The effect of Weiyanyin on the activity of SOD and the content of MDA in gastric mucosa of experiment bile reflux gastritis rats.The dividing groups and medication were same to the above,and 8 rats in every group. Reflux liquid was given to each group through the means of watering stomach except for normal group according to 15ml/kg avoirdupois,once a day, last for 35 days. Each group was fast for 24hs after watering stomach for the last time, then the head of rat was cut off to obtain stomach from abdomen, and stomach was cut open along stomach big curve, washed by physiological saline, and bloted up by filter papers. Mucosa was scraped up by reamer and weighted, and 10% tissue plasma was prepared in ice bath, which was centrifugated immediately for taking serum (3000 rs/min, 15 mins) and was kept in temperature of under -20℃to examine SOD and MDA.The activity of SOD and content of MDA in gastric mucosa were examined by contrasting-color method, and the results were revised by content of protein in tissue.Part 3: The effect of Weiyanyin on the content of TNF-α,IL-8,ICAM-1 in gastric mucosa of experiment bile reflux gastritis rats.The dividing groups and medication were same to the above,and 8 rats in every group. Reflux liquid was given to each group through the means of watering stomach except for normal group according to 15ml/kg avoirdupois,once a day, last for 35 days. Each group was fast for 24hs after watering stomach for the last time, then the head of rat was cut off to obtain stomach from abdomen, and stomach was cut open along stomach big curve, washed by physiological saline, and bloted up by filter papers. Mucosa was scraped up by reamer and weighted, and 10% tissue plasma was prepared in ice bath, which was centrifugated immediately for taking serum (3000 rs/ min, 15 mins) and was kept in temperature of under -20℃to examine the content of TNF-α,IL-8,ICAM-1.The content of TNF-αand IL-8 in gastric mucosa was examined by radiation-immunization method, while the content of ICAM-1 in gastric mucosa was examined by ELISA method, and all the results were revised by content of protein in tissue.Results:Part 1: The effect of Weiyanyin on Pathological morp- hology of experiment bile reflux gastritis rats.1 Observed by eyes: the mucosal surface of rats in normal group was smooth, and assumed pompadour; We could not see change of engorgement, fester, ulcer, and so on; The mucosal surface of the rats of model group had engorgement, fester, ulcer, and became attenuation; Compared with model group, the engorgement, fester and ulcer of Weiyanyin low-dose group and Domperidone control group was lighter, the mucosa was thicker; The mucosa of Weiyanyin high-dose group was similar to normal group.2 Observed by microscopace2.1 Fig. 1-10 showed, mucosal epidermis of rats in normal group was smooth, and cells arranged in order; The size and shape of the cells were at equal pace, the shape of the cells was like single column, the endochylema is transparent, the demarcation between endepidermis and cryptae was distinct; Few inflammatory cells could be seen; Compared with normal group, mucosal epidermis of rats in model group appeared damaged, desquamate; There were interstitial edema, anapetia, congestion and folliculus lymphaticus below the mucosa, the cell nucleus increased due to esponsiveness, and a lot of inflammatory cells could be seen; Compared with model group, epidermis cells in mucosa of Weiyanyin high-dose group, low-dose group and Domperidone control group were more abundance, and cells arranged more in order, the glandular organs lined up in order, there was not obvious cellular necrosis, a few inflammatory cells could be seen. The mucosa of Weiyanyin high-dose group was the best, which was similar to normal group.2.2 Detect inflammatory cells infiltration by semi-fixed quantity. Table 1 showed: Compared with normal group, there were more inflammatory cells in the mucosa of model group, which had a very significant difference (P<0.01); Compared with model group, there were less inflammatory cells in the mucosa of Weiyanyin high-dose group, which had a very significant difference (P<0.01); Compared with model group, there were less inflammatory cells in the mucosa of Weiyanyin low-dose group, which had a very significant difference (P<0.01); Compared with model group, there were less inflammatory cells in the mucosa of Domperidone control group, which had a significant difference (P<0.05); Compared with Domperidone control group, there were less inflammatory cells in the mucosa of Weiyanyin high-dose group, which had a significant difference (P<0.05).Part 2: The effect of Weiyanyin on the activity of SOD and the content of MDA in gastric mucosa of experiment bile reflux gastritis rats.1 The effect of Weiyanyin on the activity of SOD in gastric mucosa of experiment bile reflux gastritis rats.The table 1 showed, compared with normal group, the activity of SOD in model group was lower , which had a very notable difference (P<0.01). Compared with model group, the activity of SOD in high-dose group was higher, which had a very significant difference (P<0.01); The activity of SOD in low-dose group and Domperidone control group was also higher, which had a significant difference (P<0.05). Compared with Domperidone control group,the activity of SOD in high-dose group was higher, which had a significant difference (P<0.05); There was no marked difference of the activity of SOD among low-dose group and Domperidone control group (P>0.05).2 The effect of Weiyanyin on the content of MDA in gastric mucosa of experiment bile reflux gastritis rats.The table 2 showed, compared with normal group, the content of MDA in model group was higher, which had a very notable difference (P<0.01). Compared with model group, the content of MDA in high-dose group and low-dose group was lower, which had a very significant difference (P<0.01). Compared with model group, the content of MDA in Domperidone control group was lower, which had a significant difference (P<0.05). Compared with Domperidone control group, the content of MDA in high-dose group was lower, which had a very significant difference (P<0.01). There was no marked difference (P>0.05) of the content of MDA among low-dose group and Domperidone control group.Part 3: The effect of Weiyanyin on the content of IL-8,TNF-α,ICAM-1 in gastric mucosa of experiment bile reflux gastritis rats.1 The effect of Weiyanyin on the content of IL-8 in gastric mucosa of experiment bile reflux gastritis rats.The table 1 showed, compared with normal group, the content of IL-8 in model group was higher, which had a very notable difference (P<0.01). Compared with model group, the content of IL-8 in high-dose group and low-dose group was lower, which had a very significant difference (P<0.01). Compared with model group, the content of IL-8 in Domperidone control group was lower, which had a significant difference (P<0.05). Compared with Domperidone control group, the content of IL-8 in high-dose group was lower, which had a significant difference (P<0.05). There was no marked difference (P>0.05) of the content of IL-8 among low-dose group and Domperidone control group.2 The effect of Weiyanyin on the content of TNF-αin gastric mucosa of experiment bile reflux gastritis rats.The table 2 showed, compared with normal group, the content of TNF-αin model group was higher, which had a very notable difference (P<0.01). Compared with model group, the content of TNF-αin high-dose group was lower, which had a very significant difference (P<0.01). The content of TNF-αin low-dose group and Domperidone control group was also lower, which had a significant difference (P<0.05). There was no marked difference (P>0.05) of the content of TNF-αamong low-dose group, high-dose group and Domperidone control group.3 The effect of Weiyanyin on the content of ICAM-1 in gastric mucosa of experiment bile reflux gastritis rats.The table 3 showed, compared with normal group, the content of ICAM-1 in model group was higher , which had a very notable difference (P<0.01). Compared with model group, the content of ICAM-1 in high-dose group, low-dose group and Domperidone control group was lower, which had a very significant difference (P<0.01). Compared with Domperidone control group, the content of ICAM-1 in high-dose group was lower, which had a very significant difference (P<0.01). There was no marked difference (P>0.05) of the content of ICAM-1 among low-dose group and Domperidone control group.Conclusion:1 Weiyanyin had a good curative effect of treating bile reflux gastritis, it could improve mucosal pathological morphological of experiment bile reflux gastritis rats. So it could reduce the hurt of mucosa caused by bile reflux gastritis. 2 Weiyanyin could improve the activity of SOD, reduce the content of MDA in gastric mucosa, which restrained the oxidation of lipoprotein and resist the oxidation. So it could reduce hurt of body caused by oxidation through adjusting and improving metabolization of free radical.3 Weiyanyin could decrease the content of TNF-α,IL-8 and ICAM-1 in gastric mucosa of experiment bile reflux gastritis rats, which could reduce the number of inflammatory cells on gastric mucosa. So it could reduce the hurt of gastric mucosa caused by bile reflux gastritis.
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