Preventive Effects Of Self-made Chinese Herbal Compound Precondition On Mice Challenged With E. Coli

Objective:The study was aimed to investigate the mechanism action of immunoregulation and prophylaxis against infection diseases of the self-made Chinese herbal compound (CHC). The effects of CHC precondition on the blood physiological parameters, weight and index of thymus and spleen, Th17/Treg-related serum cytokines, key signal molecule of TLR4 signaling pathway of mice challenged with E.coli had been evaluated.Method:(1) Six mice were randomly selected and called the Control group(-72h), then 252 healthy adult mice were selected and randomly divided into six groups. Group Ⅰ、Ⅱ、Ⅲ(drug group) were respectively given 0.1mL/10g CHC (doses of 20g/kg, 10g/kg,5g/kg), Group IV (drug control group) were given 66mg/kg Astragalus polysaccharid.es, Group V (Model group) and Group Ⅵ (healthy control group)were given the equal volume physiological saline as well as the drug group, all the administration by oral for three days, once daily. Then Group Ⅰ, Ⅱ, Ⅲ, Ⅳ, Ⅴ were infected with 0.1mL E. Coli(1.7×107CFU/ml) by intraperitoneal injection at the fourth day, Group VI were injectioned 0.1ml physiological saline by intraperitoneal. (2) Blood samples were collected to measure the blood indicators by blood cells analyzer and the contents of IL-17, IL-23, IL-10, TGF-β, TLR4, MyD88, TRIF, NF-κB, P38, JNK, ERK, IRF3, TNF-a, IL-6, IFN-P in serum by ELISA at-72h, 0h,3h,6h, 12h,24h,48h,1w. Besides, thymus and spleen were collected to record for organ index at -72h, 0h,1w, and to make slices at-72h,0h,12h,1w.Result:(1)After administration of CHC for 3 days, the levels of WBC, LYM, LYM% in CHC groups were all significantly higher than those of the group without CHC administration(P<0.05). WBC. LYM, MID in CHC groups were higher than those in the infection group during 3h-24h(P<0.05), the RBC and HGB in CHC groups were higher than those in the infection group at 3h,6h,12h(P<0.05). (2) After administration of CHC, the contents of IL-17 of the medium dosage CHC group, the 1L-10 of the high dosage CHC group and the IL-17/IL-10 ratio of the medium or the low dosage CHC group were remakably higher than those of the groups without CHC administration at-72h(P<0.05). The contents of IL-10 in CHC groups were lower than that in the infection group at 3h(P<0.05). the IL-17/IL-10 ratios in CHC groups were higher than those in the infection group(P<0.01), IL-17 in the high and medium dosage CHC group were higher than those in the model group(P<0.05). The IL-23, TGF-β were lower than those in the infection group. (3)After administration of CHC, the contents of JNK, NF-κB, TNF-α in CHC groups were all significantly higher than those in the group without CHC administration(P<0.05), but P38, IL-6 were lower than those of the group not administrated with CHC(P<0.05), TRIF, IFN-β in the high dosage CHC group were higher than those in the group of non-oral CHC(P<0.01). Compared with the model group, contents of serum TLR4 in CHC groups were lower during infection(P<0.05), MyD88 were lower after 6h(P< 0.05), NF-κB were significantly lower during 3h-12h(P<0.01), P38、JNK were lower(P<0.05), the contents of ERK in high and low dosage CHC group were lower during infection(P<0.05), the contents of TNF-α and IL-6 in CHC groups were lower at 3h(P<0.01), the contents of IFN-β had no significant change(P>0.05). (4)The weights of the body, the spleen and the thymus in CHC groups were higher than those in the infection group at 1w post-infection, as well as the indexes of the spleen and the thymus (P<0.05), spleen tissue of CHC groups were lightly damaged.Conclusion:(1)Mice were infected with 0.1 mL contain 1.7×10/CFU E. Coli by intraperitoneal injection, succeeded in replicating the infection model. (2)The CHC obviously improved the mice blood physiological indexes and increased RBC, HGB, WBC. LYM and MID in the peripheral blood of mice challenged with E. coli, which improved the ability of mice to resist the E.coli infection. (3)By regulating the balance of Th17/Treg cell and the secretion of IL-17、IL-23、IL-10、TGF-p. the CHC could regulate immunity response. (4)By inhibiting the increase of NF-κB、JNK、P38、ERK、 IL-6 of TLR4 signaling pathway induced by E.coli, the CHC could reduce inflammatory response to protect the body. (5)The CHC not only promoted the growth of the thymus and spleen of mice, but also restored the histomorphology of spleen of mice challenged with E.coli.