Fuzi Lizhong Decoction On Rats With Ulcerative Colitis With Yang Deficiency Of Spleen And Kidney The Mechanism Of Action Of Treg Cell-related Factors

Objective: This study aims to explore the therapeutic effect of Fuzi Lizhong Decoction on ulcerative colitis(UC)rats with spleen and kidney yang deficiency and its effect on T Regulatory cells(Treg)related factors,In order to explore the possible mechanism of Fuzi Lizhong Decoction in the treatment of spleen and kidney yang deficiency type UC,and provide experimental basis for clinical treatment of UC.Methods: After 60 clean rats were adaptively fed for one week,they were randomly divided into a blank group(10 rats)and a model group(50 rats).The blank group was given the same amount of normal saline,and the model was first made with 15% senna2ml/100 g combined with hydrocortisone 10mg/kg to replicate the yang-deficiency model of the spleen and kidney,and the model was constantly made for 14 days.Then use TNBS/ethanol enema to replicate the UC model.Five model rats were randomly selected and sent to pathological examination to prove that the model was successful.The successfully modeled rats were randomly and equally distributed into the model group,the western medicine group,and the Fuzilizhongtang high,medium and low dose groups.Three days after modeling,gastric administration was started.The western medicine group was given a sulfasalazine suspension of 0.32g/kg·d,and the Fuzilizhongtang high,medium,and low-dose groups were treated with 3.37,6.75,and 13.50g/kg·d,respectively.The dose of gavage treatment.The blank group and the model group were given 10ml/kg of normal saline and continuous administration for 14 days.During the experiment,closely observe and record the general condition,stool condition and body weight of the rats,and evaluate the disease activity index(DAI).After the last administration,the rats were fasted with water for 24 hours,and the rats were anesthetized and laparotomy.5ml of blood from the abdominal aorta was collected.The contents of TGF-β1 and IL-10 in the rat’s serum were determined by the Elisa method.Cut the colon tissue 10 cm away from the rat anus,select a part of the colon tissue and place it in 4% paraformaldehyde solution to fix it for preparing pathological sections.After HE staining,observe the pathological morphology of each group under a light microscope;remove the remaining colon The tissue was cut and stored in a refrigerator at-80℃,and the expression of Foxp3 and STAT5 in rat colon tissue was detected by Western Blot(WB).Results:(1)General conditions of rats and DAI scores: Compared with the blank group,the DAI scores of rats in each group were significantly increased,and the difference was statistically significant(P<0.01);after treatment,compared with the model group,the DAI scores of each treatment group were reduced to different degrees,and the difference was statistically significant(P<0.05),and the Fuzilizhongtang high-dose group was the most obvious.(2)HE staining results of rat colon: the colon mucosa of the blank group is intact,the crypt structure is clear,there is no edema and inflammatory infiltration.In the model group,the colonic mucosa was eroded,exuded and congested,a large number of inflammatory cells infiltrated,the crypt structure was destroyed or changed,and the glands were arranged disorderly.Compared with the model group,the colon mucosa of each treatment group improved to different degrees.The structure of each layer was basically intact,and the crypt structure was basically intact,but the crypt surface was arranged disorderly,the number was reduced,and the crypts had different degrees of atrophy and deformation.(3)Rat serum Elisa results: Compared with the blank group,the IL-10 and TGF-β1 in the rat serum of the model group were significantly reduced,and the difference was statistically significant(P<0.01);compared with the model group,each The levels of IL-10 and TGF-β1 in the serum of rats in the treatment group increased to different degrees,and the difference was statistically significant(P < 0.05).There was no significant difference between the treatments(P>0.05).(4)WB results of rat colon tissue: Compared with the blank group,the expression of Foxp3 and STAT5 in the colon tissue of the model group was significantly reduced,and the difference was statistically significant(P<0.01);compared with the model group,each treatment Foxp3 and STAT5 in the colon tissue of rats in the group increased to varying degrees,and the difference was statistically significant(P<0.05).There was no statistically significant difference in the expression of STAT5 in colon tissue between the treatment groups(P>0.05).There were significant differences in Foxp3 expression in the attached low group,attached high school group and attached high group(P<0.01).Conclusion:(1)15% senna combined with hydrocortisone can replicate the spleen and kidney yang-deficiency model,and the TNBS/ethanol method can induce the UC model.(2)Fuzi Lizhong Decoction can improve the symptoms of UC rats with spleen and kidney yang deficiency,and can improve the inflammation of the rat colon tissue,and can repair the damaged colonic mucosa.(3)Fuzi Lizhong Decoction can up-regulate the levels of Treg-related factors IL-10,TGF-β1,Foxp3,and STAT5 to play a role in the treatment of UC of spleen and kidney yang deficiency.