| Objective:Damp-Heat syndrome of spleen-stomach (DHSS),is prevalent in Ling-nan, and generally caused by damp-heat, over-eating fatness an pungent, which may injure the spleen and stomach,is a significant task of the study on syndrome of Traditional Chinese Medicine(TCM). It is a common type of excess syndromes of spleen and stomach, it is marked with complicated clinical symptoms, longer course and it’s hard to heal completely. So in the occurrence and development of the syndrome,many factors may spur the organism and the body may counteract with the stimulation. Rat models with DHSS were established by composite element of high calorie diet, artificial environment simulation and salmonella typhimurium infection.The level of serum IFN-yand IL-4were measured by enzymelinked assay (ELISA). The expression of Bcl-2,P53were detected by SABC immunohistochemical method in50gastric mucosa tissues. Pathological changes of gastric, duodenal, colon tissues were observed using paraffin-eft section(H.E.) by the light microscope. According to the clinical features of DHSS.We observe the effects of Wangshi-Lianpu-Yin(WSLPY) on the body mass(BM), rectal temperature (RT), water and food intake, histopathological changes of the gastrointestinal mucosa, serum expression of inerleukin IL-4and interferon IFN-γ and protein expression of Bcl-2,P53.These contribute to deepen the understanding of the biological foundation and pathogenesis of DHSS, which aims to elevate the research level of DHSS.we may further reveal the biological basis and the mechanism of DHSS,and know the formation of substance of DHSS which to guide the diagnosis and treatment of DHSS.Methods:1.Chinese herbal medicine and foodstuffComposition of WSLPY:magnolia bark, golden thread, grassleaf Sweetflag rhizome, pinelliae tuber, black curd beans, Gardenia florida, Rhizoma Phragmitis. High glucose and lipid foodstuff:ordinary foodstuff mixed with12%lard and8%honey.2.GroupThe50male SD rats were randomly divided into5groups:The normal control group, Damp-Heat of Spleen-Stomach (DHSS) syndrome model group, Wangshi-lianpu-yin decoction groups of high, medium and low dose. Each group has10rats.3.Creation and modelsRat models with DHSS were established by composite element of high calorie diet, artificial environment simulation (dry bulb temperature (Td)33±0.5℃, relative humidity (rh)65±5%) and salmonella typhimurium infection. Then three WSLPY groups (high, medium and low doses) administered intragastrically with2ml/200g WSLPY for7days, the control and model group received equal volume of saline once a day.4.Indexes of observation and detectionWater drinking and food intake were recorded everyday, the body weight (BM) and rectal temperature (RT) were measured in the first, tenth, eighteenth and twenty-sixth days. After the treatment, all rats were executed, serum (500μl) was obtained after centrifuge. The level of serum IFN-y and IL-4were measured by enzymelinked assay (ELISA). The expression of Bcl-2,P53were detected by SABC immunohistochemical method in50gastric mucosa tissues. Pathological changes of gastric, duodenal, colon tissues were observed using paraffin-eft section(H.E.) by the light microscope.5.Statistic methodThe data was dealt with statistical analysis of repeated measure ANOVA and One-way ANOVA by SPSS13.0software,ranked data use the Wilcoxon Rank Sum Test,made0.05the boundary of statistical significance.Results:1.Regulation of WSLPY on the general state, food intake and water quantity changes of rats with DHSSBefore the test,all the rats were in good condition with active movements,glossy white fur,normal eating and drinking,shape stool and normal temperature. After the test,the rats in the control group were in the state of good mental health, had a shiny coat, sensitive reaction, water and food intake equilibrium, while Spleen-Stomach Damp-Heat rats gradually appeared to be sluggishness, moveless, fur lackluster, unresponsive, and took less food and water. After the treatment, the general state of rats was gradually improving, the food intake was significantly increased.The amount of water had little increased (P>0.05).2.Regulation of WSLPY on body weight (BW) and rectal temperature (RT) of rats with DHSS.Compared with the control group,body weight increased slowly (P>0.05) and rectal temperature rised rapidly (P<0.001) in the model group.All three doses of WSLPY showed a tendency to increase weight (P>0.05) and significantly reduced rectal temperature in rats with DHSS (P<0.05).3.Regulation of WSLPY on the IFN-γ/IL-4ratio in rats with DHSS.The levels of IFN-y and IL-4in model group significantly increased compared with control group(P<0.01),IFN-γ/IL-4ratio increased a little. WSLPY groups showed a significantly decreased production of both cytokines(P<0.01).There were no significant difference of IFN-γ/IL-4ratio in WSLPY groups(medium and low doses),but increased significantly in WSLPY group of high dose(P<0.05).4.Regulation of WSLPY on the Bcl-2,P53in rats with DHSS. Compared with the normal control group,there exists over expression of Bcl-2,P53in model group(P<0.05), WSLPY groups showed a decreasing tendency,but there were no significant difference(P>0.05)5.Regulation of WSLPY on the pathology of gastric, duodenal, colonic mucosal tissue.Under light microscope, in the control group,the gastric, duodenal, colonic epithelial mucosa were smooth and no erosion. In the model group, the main histological mucosal change of stomach, duodenum, colon were congestion, edema and a great of inflammation cell invasion in lamina propria and submucous layer. After the treatment with WSLPY,the inflammation in mucosa was alleviated while the gastric gland, duodenal villous and colonic epithelium were recovered.Conclusion:1. After the composite action of high glucose and lipid foodstuff,damp—heat environment and pathogenic microorganism, the spleen and stomach damp-heat syndrome animal model appeared to be the same symptoms with the clinical performance of the syndrome.Application of high calorie diet, artificial climate chamber, biological pathogenic factor can be successfully induced the animal model with damp-heat syndrome of spleen-stomach which is similar to clinical feature with the molding time.2.Over-expression of both IFN-y and IL-4indicates that rats with Spleen-Stomach Damp-Heat syndrome are in a state of violent struggle between pathogenic factor and healthy qi. It is suggested that the secretory disturbance of cell factors with Thl response tendency to drift and balancing disorder of immune network may be one of the important mechanisms in DHSS.3.There exists over expression of Bcl-2,P53in model group, there are excessive cell proliferation promote the formation of damp-heat syndrome in the spleen and stomach.4.In the model group, the main histological mucosal change of stomach, duodenum, colon were congestion, edema and a great of inflammation cell invasion in lamina propria and submucous layer,it point out rats with DHSS indicates that damp-heat syndrome of Spleen-Stomach is on the basis of morphology.5. WSLPY contains magnolia bark,golden thread,grass leaf,Sweet flag rhizome,pinelliae tuber,black curd beans,Gardenia florida and Rhizoma Phragmitis.It has functions of inhibiting inflammation and regulating balance between Thl and Th2that contributes to improve symptoms and adjust Bcl-2,P53expression in rats with DHSS,it may be one of the mechanisms that WSLPY treatment of damp-heat syndrome of spleen-stomach(DHSS). |
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