Effect Of Fuzi Lizhong Decoction Enema On Levels Of TNF-α,IL-1β And IL-6 Of Rats With UC Leaded By Yang Deficiency Of Spleen And Kidney

Objective:This dissertation attempts to investigate the action mechanism of Fuzi Lizhong decoction on UC(caused by Yang deficiency of spleen and kidney) by observing the effect of Fuzi Lizhong decoction on levels of TNF-a, IL-1β and IL-6 of rats with UC leaded by Yang deficiency of spleen and kidney. Consequently, it will do favor to the wider clinical application of Fuzi Lizhong decoction and supply experimental evidence to treat UC.Methods:(1) Modeling:60 male SD rats were randomly divided into blank group (10 rats) and model group (50 rats).Rats in the model group with 15% Senna Decoction orally 2ml/100g and subcutaneous injection of hydrocortisone 10mg/kg 15 days to create the animal model of Yang deficiency of spleen and kidney. Rats in the model group with 2,4- two nitro chlorobenzene acetic acid method to build the rat model of chronic ulcerative colitis. Rats were sacrificed to observe the part of colon tissue observed by naked eye and light microscope.(2) Treatment:42 male rats with UC(caused by Yang deficiency of spleen and kidney) are scattered into 5 groups stochastically, that is to say,10 rats belong to the model control group, and 8 rats consist of the western medicine control group. Meanwhile, groups of high-dose, medial-dose and low-dose of Fuzi Lizhong decoction have 8 rats respectively. On the other hand, there are also 8 rats regarded as normal control group. Rats in the normal control group and model control one are given enema by 3ml normal saline per day, and rats in the western medicine control group are given enema by 3ml mixed suspension of willow nitrogen sulfonyl pyridine enteric-coated metformin hydrochloride (35mg/ml), once a day. As for the other 3 groups, rats in the high-dose are offered enema by 3ml mixed suspension of high concentration (1.18g/ml) of Fuzi Lizhong decoction per day, and rats in the medial-dose are also provided clysis by 3ml mixed suspension of high concentration (0.59g/ml) of Fuzi Lizhong decoction per day, additionally, rats in the low-dose are supplied stomach lavage by 3ml mixed suspension of low concentration (0.295g/ml) of Fuzi Lizhong decoction per day. During the experiment, daily observation is about faeces, food intake, morphology, fur and movement etc, and it needs measurement of weight and anal temperature once three days. With 14 days of continuous infusion, then rats are just supplied by water (no food) in 24 hours, after that, each rat anesthetized with 3ml urethane(20%) is taken blood (5ml) from ventral aorta. Then put the blood in centrifuge with 4℃3000r/min to separate the serum, after 10mins, the centrifugation ends and put them in the refrigerator with sub-zero 20 ℃. It’s expected that ELISA would be employed to check the contents of TNF-a, IL-1β and IL-6 in serum of rats. Subsequently execute the rats by cervical dislocation and open the abdomen, and take out colon which is 4 to 10 centimeters above from the anal. Then shear and clean up the tissues, and fix the specimens by neutral formalin (10%) to make pathological sections and observe them through light microscope.Results:① Situation of faeces, food intake, morphology, fur, movement, weight and anal temperature of rats given by Fuzi Lizhong decoction is much better than the model control group.②According to observation by naked eyes and light microscope, situation of inflammation, edema and hyperemia of the colon tissue infused by Fuzi Lizhong decoction is better than that of the model control group.③Histopathological scores through light microscope demonstrate that, compared with rats in normal control group, scores of rats in model control group and those in other 4 groups of treatment are all higher than that of rats in the normal control, that is P<0.01, however, compared with model control group, scores of rats in the 4 groups of treatment are lower than that of rats in the model control group, thar is P<0.05. Measurements of TNF-a, IL-1β and IL-6 in serum of rats in the model control group and the other 4 groups of treatment are higher than those of rats in the normal control group, that is P<0.01, nevertheless, compared with the model control group, values of TNF-a, IL-1β and IL-6 in serum of rats in the 4 groups of treatment are lower than those of rats in the model control group, that is P＜0.05.Conclusion:①Fuzi Lizhong decoction is able to alleviate the following symptoms of rats with UC (caused by Yang deficiency of spleen and kidney):thin sloppy stool, mucopurulent bloody stool, less food intake, curled back, weak look, dull reaction, straight limbs, lusterless fur, less mobility. What’s more, it can ease the pathological lesion and phlogosis of rats in the model control group.②The mechanism of UC treated by enema with Fuzi Lizhong decoction could be to balance cytokines through lowering levels of TNF-a, IL-1β and IL-6, so as to be anti-inflammatory and immune.