Effects Of Ternatolide On The Expression Of IL-6, TNF-a And TGF-β In Spinal Tuberculosis Lesions In Rabbit Model

Tuberculosis is chronic treatable but the highest mortality rate ofinfectious diseases. Particularly the incidence of spinal tuberculosisrates grow with each passing day. In recent years,drug resistance andmulti frug resistant strains (multiple drugs resistant Bacilustuberculosis,MDR TB) have appeared, presenting a serious challenge toexisiting anti tuberculosis drugs, so it is very important to researchfurther the treatment of spinal tuberculosis.Studies have shown thatIL-6, TNF-a and TGF-β plays an important role in tuberculosislesions’clinical outcome,and Ranunculus ternatus lactone combinedwith antituberculous drugs can improve the efficacy,shorten the courseof treatment，have no side effects，and also can enhance the immunity ofthe patients.Objective: To explore the mechanism in the treatment of spinaltuberculosis by observing Ranunculus ternatus lactone’s effects on theexpression level of IL-6, TNF-a and TGF-β of spinal tuberculosis lesionsin rabbit model.Methods: The spinal tuberculosis model was builded by implantingtuberculosis standard strain H37Rv to unmatured rabbit L4vertebral body.20unmatured rabbits were randomly divided into4groups. A group onlyunderwent spinal tuberculosis molding, without any interventiontherapy;B group, Ranunculus ternatus lactone group, give Ranunculusternatus lactone lavage treatment after building spinal tuberculosismoldel; C group, anti tuberculosis group,give conventional anti tuberculosis drugs lavage treatment after building spinal tuberculosismoldel; D group, anti-tuberculosis+Ranunculus ternatus lactonegroup,give conventional anti tuberculosis drugs and Ranunculus ternatuslactone lavage treatment after building spinal tuberculosis moldel. Aftereight weeks, taking diseased tissues to HE staining and histologicalobservation,Real Time PCR,Western blotting to test the changes ofexpression level of IL-6，TNF-a and TGF-βof spinal tuberculosis lesionsin rabbits, before and after the treatment of Ranunculus ternatus lactoneand conventional triple anti tuberculosis drugs.SPSS16.0statisticalsoftware was applied to research further the mechanism of conventionalanti tuberculosis drugs and Ranunculus ternatus lactone spinaltuberculosisResults:After New Zealand white rabbits infected with Mycobacteriumtuberculosis H37Rv, the formation of pus cells,epithelioid cells ornecrosis foci were displayed in diseased tissues by HE staining.Thequantity of pus cells and epithelioid cells in tuberculosis lesionssignificantly reduced,after the treatment with Ranunculus ternatuslactone，conventional anti tuberculosis drugs or combinational drugsbetween them.Monitoring the relative exression of IL-6,TNF-α and TGF-βmRNA of spinal tuberculosis lesions in rabbits by RT-PCR., there aresignificantly different relative expression level in the groups oftreatment with normal saline(P<0.05) in contrast to the groups ofRanunculus ternatus lactone，conventional anti tuberculosis drugs andcombinational drugs therapy.After treatment,the relative expression ofIL-6,TNF-α and TGF-β mRNA in diseased tissue significantly reduced.Compared with the groups of treatment with Ranunculus ternatus lactone，conventional anti tuberculosis drugs, there are significantly differentrelative expression level in the groups of combinational drugs therapy(P<0.05). Among the treated groups,the lowest level of the relative exression of IL-6,TNF-α and TGF-β mRNA is the combinational drugstherapy.The results suggest that the treatment of the conventional antituberculosis drugs combined with Ranunculus ternatus lactone cansuccessfully reduce the expression level of IL-6,TNF-α and TGF-β mRNAin diseased tissue.By the way of western blotting, the expression levelof IL-6,TNF-α and TGF-β protein in diseased tissue have similar resultscomparing to the results with RT-PCR.Conclusion:1.Ranunculus ternatus lactone can effectively reduce the expression levelof IL-6,TNF-α and TGF-β in Spinal tuberculosis rabbit model,relievethe inflammatory reaction,inhibit the formation of granulation tissue inthe diseased tissues.2. Ranunculus ternatus lactone has synergistic effects on conventionaltuberculosis drugs to treat tuberculosis.