The Effect Of Actinobacillus Actinomycetemcomitans-LPS On The Expression Of Inflammation And Apoptosis-Related Genes In Tissue-Specific Macrophages

Objective:To study the relationship between periodontal disease and systemicdisease,we investigated the expression of NF-κB and the apoptosis-related genes(caspase-3, Bax，Fas, p53) in the four tissue-derived macrophages from differentorgans of rabbits with or without hyperlipidemia.The effect of Aa-LPS andinterlenkin-10on the expression of NF-κB and the apoptosis-related genes ofMonocytes/Macrophages were also investigated.Methods:The four parts of macrophages from the different organs of the rabbits withor without hyperlipidemia,which included blood mononuclear cells(Mo), alveolarmacrophages(AM), peritoneal macrophages(PM) and kupffer cells(Kc) wereharvested.These cells were randomly divided into five groups: control group, Aa-LPSgroup, Ecoli-LPS group, IL-10group and Aa-LPS+IL-10group. According todifferent groups, these cells were given Aa-LPS(1μg/ml), E.coli-LPS(1μg/ml),IL-10(0.1μg/ml).After24hours, the cells were harvested to extract protein and RNA.Finally, Western blot was used to detect the expression of NF-κB and IκB. Theapoptosis-related genes fragment were detected by real-time RT-PCR.Results:1.（1）The different parts of the macrophages which were stimulated by1μg/ml Aa-LPS showed different inflammatory changes. The expressions of NF-κB inthe AM of normal rabbits and the AM, Mo, KC of hyperlipidemic rabbits were higherwhen these cells were given Aa-LPS. But the lever of NF-κB in the PM ofhyperlipidemic rabbits were lower.（2）The trends of inflammatory changes whichwere caused by Ecoli-LPS were similar with Aa-LPS（.3）The expression of NF-κB inthe Aa-LPS+IL-10group was significantly lower than the Aa-LPS group.IL-10caninhibit inflammatory changes which were caused by Aa-LPS.2（.1）The different partsof the macrophages which were stimulated by1μg/ml Aa-LPS showed differentapoptotic changes. The Aa-LPS can promote apoptosis of AM, Mo, Kc of normal rabbit and apoptosis of Mo, AM, PM and Kc of hyperlipidemia rabbit. But it caninhibit the apoptosis in the PM of normal rabbits.（2）The trends of apoptotic changeswhich were caused by Ecoli-LPS were similar with Aa-LPS.（3）The phenomenon ofinhibition of apoptosis was not obvious when the cells were stimulated by IL-10.Conclusions:1.The different parts of the macrophages which were stimulated by1μg/ml Aa-LPS showed different inflammatory and apoptotic changes.2.The Aa-LPScould promote inflammation and apoptosis of the macrophages which were fromdifferent parts of the same rabbit.3.The effect of Aa-LPS was strengthen when theAa-LPS was used in macrophages from hyperlipidemia rabbit.4.IL-10can inhibitinflammatory changes which were caused by Aa-LPS. But the role of apoptosis wasnot obvious.