The Preparation Of Rat Model Of Yangmingfushi Syndrome And Study Of Its Pathological Mechanism

Background: Treatise on Febrile Diseases, the classic treatise, plays a significant guidance role for the clinical manifestations and medication. Yangmingfushi Syndrome- manifests as distension, fullness, dryness, sthenia and hardness - is a serial of general and synthetic symptoms such as pathogenic heat flaring inside accompanied by sthenia symptoms of stomach in the progress of febrile diseases caused by exogenous pathogenic factors. Yangmingfushi Syndrome can be seen in variety of diseases, for instance severe intra-abdominal infection, surgical abdomen, trauma, large-area burns and postoperation etc. Those diseases have the characteristic of acute onset, severity, rapid change with many complications and may cause life-threatening shock, DIC, ARDS even multiple organ failure (MODS) if not timely diagnosed or treated. However, the differentiation lacks objectives and the efficacy lacks objective evaluation criteria. Therefore, to absorb the modern and scientific methods and to strengthen the experimental study in document sorting and clinical verification is of great necessity. In the clinical research, to establish the disease animal model is an important method, and to duplicate the animal model in the research of Yangmingfushi Syndrome is also indispensable. Especially in recent years, it is urgent to build the empirical rat model of Yangmingfushi Syndrome rat, which further set a platform for integrative medicine therapy.Objective: The rat model of Yangmingfushi Syndrome, prepared through the combination of hot Chinese medicine and its feces was compared with the other two widely recognized models (made with the cecal ligation and puncture method and intraperitoneal injection of zymosan and paraffin liquid method). The concentration of plasma of rats, ET tumor necrosis factor (TNF-α) and IL-10 were measured in these three models. Use phloroglucinol method to test the d-xylose obsorption in small intestine of each group, and observe the pathological-morphological changes of the histology of the lung, small intestine, liver, colon and kidneys.Methods: Forty SD rats (20 male and 20 female rats with the weight of 180-220g) were randomly divided into four groups: control group; Chinese herbal group which was gavaged with hot Chinese herbal and its feces; ligation group which was administrated with cecal ligation and puncture; injection group which had intraperitoneal injection of Zymosan A and paraffin suspension fluid. In each group, the dosage was calculated according to each rat's weight.The Chinese herbal group: the rats were gavaged with the decoction consisting of Fu Zi, Wu Zhuyu and Gan Jiang for twelve days with the dasage of 20g/kg. The group was given fasting, intragastric administration of 2ml feces suspension fluid, bid for two days. In the mean time, 5% ethanol was drunk, changing for each day and guaranteeing its concentration. The other groups were given fasting on the same day and gavaged the same volume of water.The ligation group was given anaesthesia through intraperitoneal injection of 10% chloral hydrate on the fifth day. Then to ligate cecum root throuth McBurney's incision, puncture with 5ml syringe needle within 0.5cm from the ligation line and sew the incision. The samples were taken 24 hours later. The other groups only had McBurney's incision to dissociate cecum and then had suture without puncture and ligation.The injection group was given intraperitoneal injection of suspension liquid made by sterilized Zymonsan A powder and paraffin fluid on the fifth day with the dosage of 1.0 mg/g. The samples were taken 24 hours later. The other groups were injected the same amount of sterilized paraffin fluid.Determination of plasma D-xylose (phloroglucinol method): The rats were sacrificed one hour before the first gavage of 50g / l D-xylose solution 0.3g/kg. One hour later, the abdominal aortic blood was taken for the preparation of plasma, with phloroglucinol detection method.Plasma ET, tumor necrosis factor (TNF-α), IL-10 were detected using ELISA. The inferior vena cava blood-taking and plasma preparation were strictly administrated according to kit instructions. After the rats were killed, the liver, lung, kidney and colon were taken out and rinsed with normal saline. The slices were produced with HE staining and observed through light microscope. Results:1.General state: rats fed with hot Chinese herbs had lost weight, vertical hair, increased activity, increased water intake, reduced urine in yellow after 3 days; rats had elongation of defecation time, reduced defecation, dry feces, in peals and string beads on the first day when they were being fed with whose own stool; other rats are normal in diet, defecation, hair and behavior.2.Plasma ET level: plasma ET level in three model groups is obviously increased(P<0.05), compares to contrast group, the 3 groups doesn't have big differences(P>0.05).3.Plasma TNF-αlevel: plasma TNF-αlevel in three model groups is obviously increased(P<0.05), compares to contrast group, the 3 groups doesn't have big differences(P>0.05).4.Plasma IL-10: plasma IL-10 level in three model groups is obviously increased(P<0.05), compares to contrast group, the 3 groups doesn't have big differences(P>0.05).5.D-xylose absorption rate: plasma D-xylose level in three model groups is obviously increased(P<0.05), compares to contrast group, the 3 groups doesn't have big differences(P>0.05). 6.Histological changes: the main viscera (small intestine, liver, lung, kidneys) are all manifested as significantly inflammatory changes.Conclusions:1.The results from both hot Chinese medicine herbs feeding and own feces feeding are the same as results from the two methods of making model mentioned above and this model meets Chinese medicine theory more.2.Plasma ET, tumour necrosis factor (TNF-α), IL-10, D-xylose absorption rate, as well as some of lung, liver, kidneys, small intestine morphological changes can be used as objective diagnostic indicators of the syndrome.