The Study Of Establishment Of Animal Model Of Stomach Mucosa Lesion Of T2DM Based On Integration Of Zheng And Disease

Objective:The incidence of DM is going to show a trend of increase grandly on a global scale, and the gastrointestinal is one of the common complications of DM, and the health of humanity is going to be threaten.and the gastrointestinal is one of the common complications of DM.It has restricted the spread and use of the traditional Chinese medicine and herbal owing to subjectivity of Bianzheng of clinical of TCM (diagnose based on differentiation of symptoms and signs).This experiment aims at study of the method to establish the Integration-of-Zheng-and-Disease model of animal of stomach mucosa harm of T2DM, based on the theory of Differentiating Diagnosis of Symptoms and Signs from the holistic perspective, and the relative traditional Chinese medicine were used to cure, and the information of diagnosis of four methods of TCM were collected in order to Objectify with quantum calculation through systemic DDSS(Differentiating Diagnosis of Symptoms and Signs). The diversification of expression of level on MUCl,COX-1,COX-2,cNOS and iNOS of tissue of stomach of rats of gastric mucosal lesion of T2DM were investigated,and the time-window on Qi-deficiency of spleen and the relative effect of traditional Chinese medicine by use of rats of T2DM (Xiaoke caused by overeating or improper diet) were observed for the sake of seeking the rule about pathogenesis of vertical direction of gastric mucosal lesion of T2DM, to achieve the purpose that "to treat a disease by focusing on the root "。Methods:The number of 50 male experimental rats of SPF-SD were classified into two groups by random method:model group and blank group, and the former was fed with forage of high sugar and high fat and the latter was fed with common fodder of full level.The model group received liquid of 1% STZ at 35 minigram per kilogram, and the blank group received isodose buffer solution of sodium citrate by intraperitoneal injection in the 5th week of experiment. On random blood glucose monitoring after in 72h, the rats not only had the symptoms of polydipsia, polyphagia and polyuria but also had 3 times of the data of random blood sugar greater than 16.7 mmol/L, which were selected as animal model of T2DM, and then the alcohol of 30% was used in model group and the isodose physiological saline water was used in blank group by intragastric administration once per week for 4 weeks continuously.The rats that were selected into animal model of T2DM were classified into five groups by randomized block design:model group and Buzhong Yiqi pill group and Jinkui Shenqi pill group and a group fed with Buzhong Yiqi pill and Jinkui Shenqi pill. The isovolumic physiological saline was used in blank group and model group, and the corresponding traditional Chinese medicine were used in each medicinal-treated group simultaneously by intragastric administration after the 6th week of experiment, but the Buzhong Yiqi pill was used at first and then the Jinkui Shenqi pill was used when the rats were manifesting the syndromes and signs which were characterized by kidney-deficiency of TCM in the group that was fed with Buzhong Yiqi pill and Jinkui Shenqi pill once a day for 12 weeks. The level of random blood glucose of tail tip and glucose tolerance were collected to monitor regularly.The fasting blood glucose, INS,GLP-1,SOD,MDA and NO of serum were tested, and then the insulin relative index was calculated. The HE slices of pathology about heart, liver, spleen, kidney, stomach, intestine and pancreas were made, and the test of PcrCOX-2 and PcriNOS of the immunodetection of stomach were made. The relative test about differentiation of symptoms and signs of four diagnostic methods of TCM were used on and after the set of the model of rats of T2DM, in order to differentiate and analyze symptoms and signs and once per two weeks for 12 weeks continuously.Results:1) The results of observation of light microscopic of HE slices of pathological. First, stomach:it is normal on blank group; the pathological changes of model group in correspondence with rats of gastric mucosal lesion of T2DM, and the formation of ulcers can be seen, and the defect of mucosa direct in tunica muscularis can be seen; and the others are like blank group’s.Second, heart:it is normal on blank group; it had pathological changed on model group; the pathological changes of the group that was fed with Buzhong and Jinkui and Buzhong group like model group’s; the pathological changes of Jinkui group likes blank group’s. Third, kidney:it is normal on blank group; it had pathological changed on model group; the group that was fed with Buzhong and Jinkui, Jinkui group and Buzhong group like model group’s, but there is a mild edema Of epithelial cell of renal tubules on Jinkui group. Fourth, pancreas:it is normal on blank group; it had pathological changed of model group in correspondence with rats of gastric mucosal lesion of T2DM; there are pathological changes of different degrees of pancreatic tissues on the all treatment groups,but the pathological changes of Jinkui group likes blank group’s.Fifth, there are not changes of morphology of tissue of liver, spleen, and small intestine.2) The results of 1st of experiment. First, compared with blank group, each of the test of random blood glucose, glucose tolerance, and index of IR, ISI and insulin secretion function of model group had statistical significance (P<0.05), but the insulin hadn’t statistical significance (P>0.05). Second, compared with blank group, the MDA of model group had statistical significance (P<0.05). Third, they were always normal on blank group in the finished of experiment and in the the early terms of modeling success; the symptoms and signs of model group were in correspondence with modeling success of animal model of stomach mucosa lesion of T2DM, and had aggravated over the course of the experiment. Fourth, it was Qi-excess Zheng in the early terms of modeling success, and appearance Qi-deficiency Zheng of spleen in the 11th weeks.3) The results of 2nd of experiment. First, the Buzhong group had the best effect lowering the fasting blood glucose in the all groups, and compared with model group had statistical significance (P<0.05).Second, the pathogenesis of gastric mucosal lesion had a certain extent relation to gastric mucosa protein, and the traditional Chinese medicine had effect, and the Jinkui group had the best effect; the Jinkui group compared with blank group, it had statistical significance (P<0.05). Third, compared the inhibition rate and activity of SOD,INS,GLP-1 and NO of serum,and the PcrCOX-2 of Western-blot test of the stomach hadn’t statistical significance among block of groups and groups, and between groups all (P>0.05). Fourth,it was Buzhong and Jinkui group, Buzhong group and Jinkui group about the nose, proboscis, signs of tongue and the undertint of paws from light to heavy in the all treatment groups. Fifth,the signs and symptoms of yin-deficiency were manifested in the whole course of disease, and blank group compared with model group, Buzhong group, Jinkui group and Buzhong and Jinkui group had statistical significance (P <0.05).The Buzhong pill used in the early term of the disease was effective, and compared with model group, Jinkui group and Buzhong and Jinkui group had statistical significance (P<0.05).The degree of Yin-excess-and-deficiency can be affected by weight of rats and had statistical significance (P<0.05).Sixth, the signs and symptoms of Weire were manifested in the whole course of disease, and the all vertical comparison respective of Jinkui group and model group had statistical significance (P<0.05). The degree of Weire can be affected by weight of rats and had statistical significance (P<0.05). Seventh, the Qisheng Zheng was presented in the early time of course of disease;the Pianqi Xuzheng (deflection of pattern of Qi-deficiency) was transient presented in the middle time of course of disease; the Qisheng Zheng or Pianqi Shengzheng was presented in the latter part of course of disease. It was inexpedient that the lager quantity of Jinkui pill be used in the early time of course of disease only, and the vertical comparison of Jinkui group had statistical significance (P<0.05). Eighth,it was fundamentally normal about the degree of Xue-excess-and-deficiency in the whole course of disease, and the vertical comparison of model group had statistical significance (P<0.05)Conclusion:The experiment succeeds in the establishment of animal model of stomach mucosa lesion of T2DM based on the theory of Integration of Zheng and Disease(Xiaoke caused by overeating or improper diet), and its rate of success was 60%, and the rate of success of animal model of T2DM was 81.25%. It was Qi-excess pattern in the early terms of course of disease, and the Qi-deficiency of spleen was presented in the 11th weeks of experiment study. The oxygen-derived free radicals manifested heightened in the rats of stomach mucosa lesion of T2DM with metabolic abnormalities, and the pathogenesis of gastric mucosal lesion in relation to gastric mucosa protein to some degree, maybe it was related to excessive secretion of COX2 and secretion not enough of iNOS. The traditional Chinese medicine had effect,and the Buzhong and Jinkui group had the best effect.TO some degree, the results of viscera index indicate that the vital organs had lesions include liver, the spleen, the pancreas and the kidneys. It was Buzhong and Jinkui group, Buzhong group and Jinkui group about the nose, proboscis, signs of tongue and the undertint of paws from light to heavy in the all treatment groups successively. The degree of Blood-excess-and-deficiency was basicly nomal, and both manifested symptoms and signs of Yinxu and Weire, and the rats of stomach mucosa lesion of T2DM manifested symptoms and signs of Qisheng in the whole course of disease, and accompanied by symptoms and signs of blood stasis of varying degrees. The degree of Yin-excess-and-deficiency and Weire can be affected by the weight. The weight can be an indicator of the speciency symptoms and signs of degree of Yin-excess-and-deficiency and Weire respectively. The inhibition rate and activity of SOD, and INS.GLP-1 and NO of serum, and the PcrCOX-2 of immunodetection of stomach cannot be affected by the weight.The Yinxu-Zaore(Yin-deficiency results in hyperactivity of yang) was the root of pathogenesis of TCM of the rats of stomach mucosa lesion of T2DM (Xiaoke caused by overeating or improper diet), and the rule about pathogenesis of vertical direction of the rats of stomach mucosa lesion of T2DM was Weire lead to Yin-deficiency, and then Zaore-growing inward, and consequently the lung was harmed by Zaore, and then the kidneys and the spleen were harmed, and accompanied by symptoms and signs of blood stasis of varying degrees, resulting in harm of Sanjiao or deficiency of Yinyang, or even death due to dropout of yang from depletion of Yin without effects of curing.