Splitting study of mechanism of Jianpi-Huazhi pill in treatment of ulcerative colitis based on TLR-4/MyD88/NF-κB pathway
Objective:To establish ulcerative colitis(UC)rat model of spleen deficiency and dampness,and to explore effect of Jianpi-Huazhu pill and its different deconditions on TLR-4/MyD88/NF-κB pathway.Methods:A total of 64 Wistar rats were ran-domly divided into normal control group(n=8)and model group(n=56).UC rat model was established by traditional Chinese medi-cine syndrome combined with ethanol-2,4,6-trinitrobenzene sulfonic acid enema method.After successfully modeling,rats were ran-domly divided into model group(M),Mesalazine group(A),Jianpi-Huazhi pill group(B),Jianpi-Qinghua decoction group(C),Jianpi-Qinghua-Huoxue decoction group(D),Jianpi-Qinghua-Daozhi decoction group(E)and Qinghua-Daozhi-Huoxue decoction group(F),with 8 rats in each group,given corresponding drugs by gavage for 4 weeks.At end of course of treatment,all rats were sacrificed and samples were taken.Changes of body weight,disease activity and pathological changes of colon were observed.TNF-α expression in colon tissue was detected by ELISA and immunohistochemistry.RT-PCR and Western blot were used to detect mRNA and protein expressions of TLR-4,MyD88 and NF-κB in rat colon tissue.Results:Rats weight in Jianpi-Huazhi pill group,Jianpi-Qinghua decoction group,Jianpi-Qinghua-Huoxue decoction group and Jianpi-Qinghua-Daozhi decoction group were obviously higher than that of model group(P<0.01),all treatment groups could obviously improve UC model rat pathological damage and disease activity index in colon tissue,and Jianpi-Huazhi pill group and Jianpi-Qinghua-Huoxue group showed better efficacy.All treatment groups could inhibit mRNA and protein expressions of TLR-4,MyD88 and NF-κB in colon tissue(P<0.001).Mesalazine group,Jianpi-Huazhi pill group,Jianpi-Qinghua-Daozhi group and Qinghua-Daozhi-Huoxue group were significantly better than Jianpi-Qinghua group and Jianpi-Qinghua-Huoxue group in reducing NF-κB mRNA and protein expressions(P<0.05).All groups could reduce expres-sion of TNF-α in colon tissue(P<0.001),and there was no significant difference between groups(P>0.05).Conclusion:Jianpi-Huazhi pill and its various deconditions may play roles through TLR-4/MyD88/NF-κB pathway.Among them,Coptid-Rhizoma coptifo-lia,stews woody incense,Fengwei herb and gung ginger may be core drugs of this recipe,which has certain guiding significance for clinical practice.