| Chronic superficial gastritis (CSG) is a kind of chronic superficial inflammation in gastric mucosa and which is caused by a variety of reasons.It is one of the digestive diseases with higher incidence. The clinical symptoms of CSG include: chronic irregular upper abdominal pain, bloating, belching, etc.Which is particularly obvious when diet is irregular. Some patients may have acid reflux, occasional vomiting, and so on. The causes of CSG include: bacteria, viruses and toxins, smoking, certain drugs, spicy food, recycling, and metabolic dysfunction, bile and duodenal juice reflux, Helicobacter pylori (HP) infection, physical and psychological factors,etc.In TCM the disease is called "Stomachache", "fullness", "noisy", etc, the causes include: stomach invaded by exogenous evils, improper diet, emotional disorders, deficiency of spleen and stomach,and so on. The location is in the stomach, and the disease has close relationship with the liver and the spleen. It can be mainly divided into: the syndrome of spleen-stomach deficiency, the syndrome of incoordination between the liver and the stomach, the syndrome of damp-heat in the spleen-stomach ,the syndrome of stomach-yin deficiency, the syndrome of blood stasis in stomach,etc.In this study, we established the Combination of Syndrome with Disease Model .We copied the disease of western medicine (chronic superficial gastritis) and the syndromes of traditional Chinese medicine (the syndrome of spleen deficiency and the syndrome of damp-heat) in order to investigate the difference of the same disease with different syndromes in energy metabolism and apoptosis aspects,so that we can investigate the essence of the syndrome of TCM.Purpose:Establish the CSG rat Combination of Syndrome with Disease Model, then investigate the difference of the CSG Rat Combination of Syndrome with Disease Model in energy metabolism and apoptosis aspects.According to the difference of the relevant indicators,we analyze the correlation of the same disease with different syndromes in energy metabolism and apoptosis aspects,so that we can provide evidence for the syndrome differentiation and treatment of TCM on the micro level.Materials and methods:1.We replicated the syndrome model and the disease model at the same time.We selected ten rats randomly as the normal control group and fed them with 2ml saline every day.We fed the other thirty rats with 5% sodium salicylate 10ml/kg·d according to the their weights.This period lasted three weeks.Three weeks later,we divided the thirty rats into three groups randomly: normal group, the simple CSG group, damp-heat CSG group and spleen deficiency CSG group.Each group was fed with 5% sodium salicylate 10ml/kg ? d according to their weights,at the same time,we had no extra intervention to the simple CSG group,we fed the damp-heat CSG group rats with 200g / L honey water and the rats can drink it freely.We also fed the rats of damp-heat CSG group 10ml/kg·d liquor every other day and fed them fat 10ml/kg·d when the day without liquor.We replicated the syndrome of spleen deficiency with laxative and irregular diet .We fed them with Xiao Cheng Qi decoction (rhubarb 12g, Magnolia 15g, citrus aurantium 9g) 10ml/kg·d in animal dose.We gave them half diet every other day and they can drink water freely.This period lasted two weeks.After the second period they were executed.2.Observe the pathological changes of rat gastric mucosa.3.Detect the content of SDH in gastric tissue by Colorimetric methods.Detect the content of Cyt-C in gastric tissue by ELISA methods.Detect the apoptosis condition of gastric epithelial cells by TUNEL methods.Observe the expression of the the rat gastric mucosa UCP2mRNA and the expression of apoptosis genes Bcl-2mRNA in gastric mucosa.Results:1.The general state of rats:damp-heat CSG group:The weight of the damp-heat CSG group grew rapidly firstly,and gradually slowing down later,they seemed sleepy and lazy with less moving, they had poor appetite and drunk less, they had dry or soft stools, their fur was loose and rough, even erected.Their rectal temperature increased comparing with the previous.spleen deficiency CSG Group: The rats of this group had loose stools and dirty crissum. They had poor appetite and less diet.They seemed haggard and apathetic with eyes half closed.They looked arched and they had slow weight gain, their rectal temperature decreased comparing with the previous. The comparative results of the increased weight: normal group> simple CSG group> damp-heat CSG group> spleen deficiency CSG group. The results of the spleen CSG group compared to the other three groups were statistically significant,Compared with the normal group and the damp-heat CSG group, P <0.01, compared with the simple CSG group, P <0.05;the comparative results of the increased rectal temperature:damp-heat CSG group> simple CSG group> normal group> spleen deficiency CSG group, in which the spleen deficiency CSG group compared with the damp-heat CSG group and the simple CSG group, P <0.01, compared with the normal group, P <0.05.The pathological changes of each group in gastric epithelial cells: normal group: The rats of this group had normal gastric mucosa with clear hierarchy and integrity epithelial structure without damage, the mucosal glands arranged in neat rows, the glandular cavities were clean and the mucous layer had no inflammatory cell infiltration;Simple CSG group: We could see mild inflammatory cell infiltration in the propria lamina of the gastric mucosa. We could see vasodilatation and congestion in the submucosa; spleen deficiency CSG group:We could see the defect, loss and necrosis of the mucosa. We could see inflammatory cell infiltration in the propria lamina of the gastric mucosa; damp-heat CSG group: We could see the defect, loss and necrosis of the mucosa. We could see more inflammatory cell infiltration in the propria lamina of the gastric mucosa. The histopathological score of the gastric inflammation in rats : The pathological score comparison of the gastric inflammation among the four groups: normal group<simple CSG group <spleen deficiency CSG group <damp-heat CSG group, the normal group compared with the other three groups :P <0.01;damp-heat CSG group compared with the spleen deficiency CSG group, P <0.05, damp-heat CSG group compared with the simple CSG group, P <0.01.2.The changes of SDH contents: damp-heat CSG group> normal group> simple CSG group> spleen deficiency CSG group: Among them, the damp-heat CSG group compared with the other three groups, there was statistical significance, compared with the normal group, P <0.05, compared with the simple CSG group and the spleen deficiency CSG group, P<0.01; the spleen deficiency CSG group compared with the other three groups, P <0.01, the normal group compared with the simple CSG group, P> 0.05.3.The changes of the cytoplasm Cyt-C content levels:spleen deficiency CSG group>damp-heat CSG group> simple CSG group> normal group. Among them, the spleen deficiency CSG group compared to the other three groups, P <0.01; the normal group compared with the simple CSG group, P <0.05, the normal group compared with the damp-heat CSG group, P <0.01; simple CSG group compared with the damp-heat CSG group, P> 0.05.4.The expression of rat gastric UCP2mRNA from strong to weak as follows: spleen deficiency CSG group> simple CSG group> normal group> damp-heat CSG group. Spleen deficiency CSG group compared with the simple CSG group and the normal group, P <0.05, and it compared with the damp-heat CSG group, P <0.01.5.The changes of the apoptosis index of rat gastric epithelial cell : spleen deficiency CSG group>damp-heat CSG group> simple CSG group>normal group. Among them, the normal group compared with the other three groups, P <0.01; spleen deficiency CSG group compared with the damp-heat CSG group, P <0.05, it compared with the simple CSG group, P <0.01; Compared to the normal group ,these three model groups had decreased expression of Bcl-2mRNA .The expression of Bcl-2mRNA in spleen deficiency CSG group is more lower than that in other groups. Compared to the other three groups the comparison result was statistically significant. Which compared with simple CSG group, P <0.05, compared with the normal group and the damp-heat CSG group, P <0.01. Conclusions:1.According to the general state observed in rats, including rats appearance, diet, body weight, urine, fur, mental state, activity, rectal temperature, etc,we found that we can establish the damp-heat syndrome and the spleen deficiency syndrome models in this way. According to the gastric pathology of the rats, we could find the features of chronic superficial gastritis. We can reproduce CSG Combination of Syndrome with Disease Model rat with this method.2.The changes of SDH content in damp-heat CSG group and spleen deficiency CSG group, reflected the diffence of damp-heat snydrome and spleen deficiency syndrome under the same disease state (chronic superficial gastritis) in energy metabolism aspect.3.The rat gastric cytoplasm Cyt-C content in spleen deficiency CSG group was significantly higher than that in the other three groups, which reflected that the increased membrane permeability in spleen deficiency CSG group might occur.Cyt-C might get into the cytoplasm from the mitochondria, and result in the increase of the cytoplasmic Cyt-C levels. Which also reflected the spleen deficiency status will result in the dysfunction of mitochondrial respiratory chain, so that the ATP was in deficiency and the energy production was in shortage.4.The expression of UCP2mRNA was significantly higher in spleen deficiency CSG group than that in the other three groups. Which suggested that the ATP generation decrease in spleen deficiency CSG group, while the expression of UCP2mRNA was lower in damp-heat CSG group,it suggested that this group can produce more ATP ,so that it can regulate energy metabolism.The result was identical with experiment two and experiment three previously,they all show there was a dysfuction of energy metabolism in spleen deficiency CSG group and there was a hyperfunction of energy metabolism in damp-heat CSG group,but the regulation mechanism of UCP2 in this experiment remained to be further studied.5.The comparative result of rat gastric epithelial cell apoptosis index :spleen deficiency CSG Group>damp-heat CSG Group> simple CSG group> normal group. damp-heat CSG Group had serious mucosal inflammatory reaction, the apoptosis index of this group increased, and spleen deficiency CSG group had gastric dysfunction, so that the apoptosis index of this group increased. According to the Bcl-2mRNA expression of the apoptosis genes ,we found that the Bcl-2mRNA expression and the occurrence of apoptosis was closely related,and they correlated negatively.6. The CSG rat in different syndromes had relavance in energy metabolism and apoptosis aspects.The dysfunction of energy metabolism and the apoptotic index showed a higher consistency, and the cytochrome C played an important role in terms of both energy metabolism and apoptosis. |
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