Effects Of Exo-endo Damp-heat Factors On Immunological Reaction On Respirovirus Induced By TLRs

Infections of respirovirus are common and frequently-occurring diseases, in which the influenza was most severe and broke out actively recent years. The occurrence of viral deseases is related to viral pathogenicity and people's immunity, but antiviral drugs had limited effects, so it's the point of researches to increase or regulate immunity. TLRs play an important role in antiviral immunity, so TLRs can be not only the new treatment of viral diseases but also new targets of the manufacture of drugs and vaccines. Respirovirus infections in south of Five Ridgs, namely Lingnan, especial the Pearl River Delta, are apt to transmit to heat syndrome and combine damp syndrome because of heat and humid climate, dense population and extraordinary diet. Haoqinqingdan decoction had significant effects on the damp-heat syndrome of Lingnan's respirovirus infections in our past experiences. By observing the effects of exo-endo damp-heat factors on immunological reaction on respirovirus induced by TLRs and the intervention effect of Haoqingqindan decoction and its decomposed recipes, We investigated the characteristics of damp-heat syndrome in immunological reaction induced by TLRs and anti-viral mechanism and targets of Haoqingqindan decoction, and hoped to provide experimental and clinical evidences for further study of Chinese medicine therapy of viral diseases.ObjectiveTo observe endogenous and exogenous damp-heat factors'effects on the expressions of TLR-7,NF-κB,MyD88 in mice infected by influenza virus through stimulating endogenous and exogenous damp-heat condtions(a richly fatty and sweet diet, heat and humid environment). And to investigate the characteristic of damp-heat syndrome of respirovirus infections in immunological reaction induced by TLRs, while the differences of NF-κB,MyD88 between damp-heat group and control group in clinical experiment were concerned coordinately. We also wanted to offer supplements and evidences for those before by study on the effects of Haoqingqindan decoction and its decomposed recipes on the expression of TLR-7,NF-κB,MyD88 in mouse liver tissue and patients'surum and investigate its mechanism and targets.Methods1 Experimental researches:①to study the effect of exo-endo damp-heat factors on immunological reaction on respirovirus induced by TLR-7:40 mice of BALB/c were devided into 4 group randomly (control group, damp-heat group, model group, viral group), then observed the symptom and sign of mice. After executions, lung index and the expression of Influenza virus, TLR-7, NF-κB and MyD88 in lungs of mice were detected by RT-PCR.②to study the intervention effect of Haoqingqindan decoction and its decomposed recipes on immunological reaction on respirovirus induced by TLR-7:115 mice of BALB/c were devided into 8 group randomly (control group, model group, Ribavirin group, HQQD group, decomposed 1 group, decomposed 2 group, decomposed 3 group, decomposed 4 group), then observed the symptom and sign of mice. After executions, lung index and the expression of Influenza virus, TLR-7, NF-κB and MyD88 in lungs of mice were detected by RT-PCR.2 Clinical research:To collect 30 patients as damp-heat group, who met the diagnosis of Western medicine and damp-heat syndrome of Chinese medicine, while 15 health volunteers were collected randomly as control group. Symptoms and signs and the expression of NF-κB and MyD88 in serum were detected by ELISA before and after treantment.Results1 Experimental researches1) The mortality of model group was a little higher than viral group. The lung index of model group is a little lower than viral group, but the difference wasn't significant (P>0.05). Under light microscope, there were severe inflammatory cell infiltrations in bronchioles, congestion in blood capillaries of pulmonary interstitium and effusion in alveolar space in model group. Pathological changes in model group were more severe than in viral group. Under electron microscope, Pathologic changes in model group was about the same as in viral group, but there were a large number of lipid droplets in alveolar epithelium of model group.2) The expression of influenza viruses in model group was a little higher than in viral group, but the difference wasn't significant (P>0.05).3) The mean levels of TLR-7,MyD88,NF-κB in model and viral group was higher than those in control group, but the difference of TLR-7,MyD88 wasn't significant while the difference of NF-κB was significant; The mean levels of TLR-7,MyD88,NF-κB in model group increased to a certain extent than those in viral group, and the difference of NF-κB was significant. The increasing extents of TLR-7,MyD88,NF-κB weren't parallel.4) Haoqingqindan decoction and its decomposed recipes could inhibit pathological changes, replicate of influenza viruses and decrease the expression of MyD88,NF-κB to a certain extent, in which Haoqingqindan decoction had the best effect.2 Clinical researchThe expression of MyD88,NF-κB in Damp-heat group was higher than in control group, and the difference was significant. After treatment of Haoqingqindan decoction, the expression of MyD88,NF-κB in Damp-heat group increased, and the difference was significant comparing with before treatment. Conclusions1 Those results of the first experimental research indicated that the increasing of the expression of TLR-7,MyD88,NF-κB is relation to the infections of influenza viruses, but the interventions of exo-endo damp-heat factors also work on it, which to stimulate the secretion of a large number of inflammatory cells factors (such as TNF-α,IL-12 and so on). Then over reaction of inflammatory damaged the normal tissues and functions, which led to severe pathogenetic condition and high mortality. It's also the evidence for those above that the expression of MyD88,NF-κB in damp-heat group increased comparing with control group in clinical test and the difference was significant.2 In experimental research, the increasing extents of TLR-7,MyD88 NF-κB weren't parallel, we considered that there is likely to be three reasons:①viral virulence:Virulence was weak because of lower viral concentration, so the expression was lower.②course of disease:TLR-7 and MyD88 had been exhausted in later period.③another signal passage:there is another signal passage to stimulate NF-κB,too. Pertinent evidences need further researches.3 Haoqingqindan decoction and its decomposed recipes could inhibit pathological changes, replicate of influenza viruses and decrease the expression of MyD88,NF-κB to a certain extent, which indicated that the expression of TLR-7,MyD88,NF-κB can be decreased by clearing heat and eliminating dampness, regulating the stomach and Qi. We inferred that the interventions of exo-endo damp-heat factors probably affect the levels of the expression of TLR-7,MyD88,NF-κB. So to remove the exo-endo damp-heat factors can improve the series of pathogenical changes induced by the intervention, which maybe provide a clue or breakthrough point for further study the therapeutic mechanism of Haoqingqingdan decoction on the damp-heat syndrome of viral disease in Lingnan.