| Purpose:Using molecular biology techniques,this study through the in vivo experiment,observation,and gastric reflux kang on treating bile reflux gastritis model rats general situation,gastric mucosa pathological histology,plasma gastric dynamic element(MOT)expression and the influence of the content;Observation and gastric reflux kang on treating bile reflux gastritis model rat gastric mucosa interleukin 6(IL-6),interleukin 8(IL-8)expression and the influence of the content;Observation and gastric reflux kang on treating bile reflux gastritis model rats serum gastric gastrin-releasing(GAS),gastric mucosa prostaglandin E2(PGE2)expression and the influence of the content;Observation and gastric reflux kang on treating bile reflux gastritis model rat gastric mucosa p38 MAPK protein and gastric mucosa cell cox-2 mrna expression,in order to and gastric reflux treatment of bile reflux gastritis has a more comprehensive understanding of fully,for traditional Chinese medicine in the treatment of bile reflux gastritis provide new theoretical basis.Material and method:60 rats(SPF)were randomly divided into 6 groups,respectively,blank control group,model control group,positive drug(ding Lin)in the control group,and the gastric reflux,low dose group,and gastric reflux dose group,and gastric reflux in kang kang high dose group,each group of 10.Preparation reflux liquid,in addition to the blank control group,the rest of the group are used to build experimental gastric bile reflux gastritis rats model,1 times a day,five weeks in a row.Groups of rats after since building from 2 weeks to 5 Zhou Zao die end,all the drug lavage 1 times a day.After the last time to fill the stomach,the groups of rats fasting,24 h and execution,application of HE staining combined with semi quantitative detection observation and gastric reflux of bile reflux gastritis,gastric mucosa pathological histology in rats;Using immunohistochemical(IHC)method to detect the distal gastric mucosa tissues of IL-6,IL-8,MOT,GAS,PGE2 expression level;Enzyme linked immunosorbent(ELISA)method to detect the rat gastric mucosa in serum IL-6,IL-8,GAS,the content of PGE2,plasma MOT,observation and gastric reflux kang on treating bile reflux gastritis model rat gastric mucosa serum IL-6,IL-8,GAS,the influence of PGE2,plasma MOT expression and content;Using Western blot method to detect the distal gastric mucosa tissues p38 MAPK protein expression,using real-time fluorescent quantitative PCR method to detect the rat gastric mucosa cell cox-2 mrna expression,observation and gastric reflux kang on treating bile reflux gastritis model rat gastric mucosa p38 MAPK protein and gastric mucosa cell cox-2 mrna expression.Results: General condition: the influence of the blank control group rats fur color is white,colour and lustre is light,responsive,flexible,active foraging for food,food intake and water quantity is normal,defecate spherical;Depression model group rats,fur color yellow,lacklustre,unresponsive,eyes without the god,lazy to move,eating and drinking to reduce gradually,body emaciated,defecate pond.And gastric reflux,high,medium and low dose group and ding Lin group rats generally in good condition,fur fade luster,food intake and drinking also increase gradually,weight increase gradually,but in and gastric reflux,high dose group improved the most obvious,and gastric reflux,middle dose group.1.Gastric mucosa pathological histology observations Model group rats with gastric mucosa appeared obvious damage,fall off,submucosal numerous inflammatory cells infiltration,most of the rat gastric mucosa pathological tissue in atypical hyperplasia and intestinal metaplasia.Compared with model control group,and gastric reflux,high,medium and low dose group and gastric mucosa injury in rats ding Lin control group is lighter,and gastric reflux kang,low dose group and ding Lin gastric mucosa is still visible part of the inflammatory cells infiltration,but were significantly reduced pathology in the blank group,some phenomenon of intestinal metaplasia epithelium.And gastric reflux,high dose group have almost the same shape,the size of the gastric mucosa gland epithelium arrangement is neat,rarely found obviously congestion and necrosis,with a small amount of inflammatory cell infiltration in the mucosa,intestinal metaplasia,seldom appears almost blank group of gastric mucosa in rats.2.Immunohistochemical method to detect the results(IHC)(1)MOT expression level Compared with the blank control group,model group MOT expression in gastric mucosa tissue of rats decreased obviously,statistically significant(P < 0.01);Compared with model control group,and gastric reflux,high,medium and low dose group and ding Lin rat gastric mucosa tissue MOT expression were significantly higher(P < 0.01);Compared with ding Lin control group,and gastric reflux,high dose group of rat gastric mucosa tissue MOT expression is significantly higher than ding Lin in the control group(P < 0.01).(2)IL-6,IL-8 expression level Compared with blank control group,model group rats IL-6 expression in gastric mucosa tissue levels were significantly elevated,statistically significant(p < 0.01);Compared with model control group,and gastric reflux,high,medium and low dose group and ding Lin IL-6 in rat gastric mucosa tissue expression levels were significantly reduced,statistically significant(p < 0.01);Compared with butyl Lin control group,and gastric reflux,low dose group of IL-6 in rat gastric mucosa tissue expression level increased significantly(p < 0.01),and gastric reflux,high,medium dose group of IL-6 in rat gastric mucosa tissue expression level,though there was no significant difference(p > 0.05),numerical compared with have a downward trend.Compared with blank control group,model group rats IL-8 expression in gastric mucosa tissue levels were significantly elevated,statistically significant(p < 0.01);Compared with model control group,and gastric reflux,high,middle dose group and ding Lin IL-8 expression in rat gastric mucosa tissue levels were significantly reduced,statistically significant(p < 0.01);Compared with ding Lin control group,and gastric reflux,low dose group of rat gastric mucosa tissues IL-8 expression level increased significantly(p < 0.01),and gastric reflux,high,middle dose group and the comparison of numerical has a downward trend,but no significant difference(p > 0.05).(3)GAS,PGE2 expression level Compared with blank control group,model group rats gastric mucosa tissues GAS expression levels were significantly reduced,statistically significant(p < 0.01);Compared with model control group,and gastric reflux,high,medium and low dose group and ding Lin controls GAS expression in rat gastric mucosa tissue levels were significantly elevated, statistically significant(p < 0.01 or p < 0.05);Compared with butyl Lin control group,and gastric reflux kang GAS in high dose group of rat gastric mucosa tissue expression level increased significantly(p < 0.01).Compared with blank control group,model group rats obviously decrease PGE2 expression in gastric mucosa tissue level,statistically significant(p < 0.01);Compared with model control group,and gastric reflux,high,medium and low dose group and ding Lin controls the level of PGE2 expression in gastric mucosa tissue of rats was significantly elevated,statistically significant(p < 0.01);Compared with ding Lin control group,and gastric reflux,high dose group of PGE2 in gastric mucosa tissue in rat expression level increased significantly(p < 0.01).3.Enzyme-linked immunosorbent(ELISA)method is used to test results(1)Content of plasma MOT Compared with blank control group,model group rats gastric mucosa tissue plasma MOT content were significantly reduced,statistically significant(p < 0.01);Compared with model control group,and gastric reflux,high,middle dose group and ding Lin rat gastric mucosa tissues were significantly elevated plasma MOT content,statistically significant(p < 0.01);Compared with ding Lin control group,and gastric reflux,low dose group of rats with gastric mucosa plasma MOT content significantly decreased(p < 0.01),and gastric reflux,high,middle dose group of rats with gastric mucosa plasma MOT content has a rising trend,but there was no significant difference(p > 0.05).(2)IL-6,IL-8 levels Compared with blank control group,model group content of the IL-6 in rat gastric mucosa tissue increased significantly,statistically significant(p < 0.01);Compared with model control group,and gastric reflux,high,medium and low dose group and ding Lin IL-6 content in rat gastric mucosa tissues were significantly reduced,statistically significant(p < 0.01);Compared with ding Lin control group,and gastric reflux,high dose group of rat gastric mucosa tissues significantly lower content of IL-6,and gastric reflux,low dose group of IL-6 levels increased significantly(p < 0.05).Compared with blank control group,model group rats gastric mucosa tissues IL-8 levels increased significantly,statistically significant (p < 0.01);Compared with model control group,and gastric reflux,high,medium and low dose group and ding Lin IL-8 content in rat gastric mucosa tissues were significantly reduced,statistically significant(p < 0.01);Compared with ding Lin control group,and gastric reflux,low dose group of rat gastric mucosa tissues IL-8 levels increased significantly(p < 0.01)(3)the content of GAS?PGE2 Compared with blank control group,model group GAS content in the rat gastric mucosa tissues were significantly reduced,statistically significant(p < 0.01);Compared with model control group,and gastric reflux,high,medium and low dose group and ding Lin GAS content in the control group rats gastric mucosa tissues were significantly elevated,statistically significant(p < 0.01,p < 0.05);Compared with ding Lin control group,and gastric reflux,low dose group of GAS content in the rat gastric mucosa tissue decreased obviously(p < 0.01),and gastric reflux,high,middle dose group and the comparison of numerical has a rising trend,but no significant difference(p > 0.05).Compared with blank control group,model group PGE2 content in rat gastric mucosa tissues were significantly reduced,statistically significant(p < 0.01);Compared with model control group,and gastric reflux,high,medium and low dose group and ding Lin PGE2 content in rat gastric mucosa tissues were significantly elevated,statistically significant(p < 0.01);Compared with ding Lin control group,and gastric reflux,high dose group of PGE2 levels in rat gastric mucosa tissue increased significantly(p < 0.01).4.RT-PCR method groups of test results COX-2m RNA expression level.Compared with blank control group,model group rats gastric mucosa tissues cox-2 mrna expression level increased significantly,statistically significant(p < 0.01);Compared with model control group,and gastric reflux,high,medium and low dose group and ding Lin control rat gastric mucosa tissues cox-2 mrna expression levels were significantly reduced,statistically significant(p < 0.01);Compared with butyl Lin control group,high and low dose group and gastric reflux kang rat gastric mucosa tissues cox-2 mrna expression level increased significantly(p < 0.01,or p < 0.05).5.Western Blot method groups of test results P38 MAPK protein expression levels Compared with blank control group,model group rats gastric mucosa tissue p38 MAPK expression levels were significantly elevated,statistically significant(p < 0.01);Compared with model control group,and gastric reflux,high,medium and low dose group and ding Lin p38 MAPK protein expression level in rat gastric mucosa tissues were significantly reduced,statistically significant(p < 0.01);Compared with ding Lin control group,and gastric reflux,high dose group of rat gastric mucosa tissues p38 MAPK protein expression levels significantly decreased(p < 0.05).Conclusion: 1.He wei fan liu kang can significantly improve the experimental bile reflux gastritis,gastric mucosa pathological histology in rats,improve gastric dynamic element(MOT)content in plasma,better improve gastric mucosa damage,promote digestion period between gastric emptying and reflux liquid removal.2.He wei fan liu kang can significantly reduce the experimental bile reflux gastritis rats gastric mucosa inflammatory cytokines IL-6,IL-8 levels,inhibit rat gastric mucosa inflammatory cytokine expression of IL-6,IL-8,reduce reflux liquid inflammatory damage of gastric mucosa,so as to better protect the gastric mucosa.3.He wei fan liu kang can significantly increase experimental bile reflux gastritis rats stomach gastrin-releasing(GAS)content in serum,promote gastric secrete element(GAS)in the serum of rats,better improve gastric secretion and motor function,promote restoration of the gastric mucosa;And gastric reflux kang can significantly improve the experimental bile reflux gastritis rats gastric mucosa prostaglandin E2(PGE2)levels,promote rat stomach mucosa the expression of prostaglandin E2(PGE2),enhance the gastric mucosal immunity,cell protection.4.He wei fan liu kang can significantly inhibit experimental bile reflux gastritis rats gastric mucosa p38 MAPK protein and gastric mucosa cell cox-2 mrna expression,reduce the flow fluid inflammatory damage of gastric mucosa,prevent BRG model rat gastric mucosa of intestinal metaplasia and atypical hyperplasia of precancerous lesions occur. |
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