青藤碱对牙龈卟啉单胞菌脂多糖诱导的人牙龈成纤维细胞IL-8表达的影响  

Effect of sinomenine on IL-8 expression in porphyromonas gingivalis lipopolysaccharide-stimulated human gingival fibroblasts

作　　者：王春[1] 向学熔[1] 杨明聪[1] 陈芳淳[1] 

机构地区：[1]重庆医科大学附属口腔医院牙周科,重庆401147

出　　处：《第三军医大学学报》2012年第19期1972-1976,共5页Journal of Third Military Medical University

摘　　要：目的探讨青藤碱对牙龈卟啉单胞菌脂多糖诱导的人牙龈成纤维细胞表达IL-8的影响。方法用不同浓度的青藤碱(25、50、100μmol/L)预处理24 h后,再加入1μg/ml的脂多糖(LPS)处理人牙龈成纤维细胞。采用RT-PCR、ELISA法检测青藤碱对脂多糖诱导细胞表达IL-8的影响;用Western blot法检测青藤碱对NF-κB核转位及IκBα降解的影响。加入NF-κB抑制剂Bay11-7085 10μmol/L、TPCK 20μmol/L预处理细胞24 h后,再用LPS刺激细胞4 h,采用ELISA法检测NF-κB抑制剂对脂多糖诱导的IL-8表达的影响。结果青藤碱剂量依赖性地抑制脂多糖诱导的人牙龈成纤维细胞中IL-8 mRNA及蛋白表达(P<0.05);与脂多糖组比较,青藤碱基本上把NF-κB阻滞在细胞质中,而细胞经脂多糖处理1 h后,大部分NF-κB被转运到细胞核中,经100μmol/L青藤碱预处理细胞的细胞质中NF-κB含量为脂多糖组的2倍;脂多糖处理细胞1 h后,50%的IκBα降解,但经青藤碱预处理细胞后,呈浓度依赖性抑制脂多糖诱导IκBα的降解(P<0.05)。50μmol/L的青藤碱使脂多糖诱导IκBα的降解恢复到正常水平;脂多糖处理组IL-8表达量为对照组的2.4倍,但加入NF-κB抑制剂Bay11-7085(10μmol/L)、TPCK(20μmol/L)后IL-8蛋白的表达量分别下降为对照组的1.8倍和1.6倍。结论青滕碱可能通过阻断NF-κB的核转位以及IκBα的降解来抑制牙龈卟啉单胞菌脂多糖诱导的人牙龈成纤维细胞IL-8的合成。study the effect of sinomenine （Sin） on IL-8 expression in porphyromonas gingivalis lipopolysaccharide （LPS）-stimulated human gingival fibroblasts （HGF）. Methods HGF were pretreated with Sin at the concentrations of 25,50 and 100 μmol/L for 24 h and then with 1 μg/ml LPS which was added into Sin. Effect of Sin on LPS-mediated IL-8 expression in HGF was detected by RT-PCR and ELISA, respectively. Effect of Sin on NF-KB translocation and IB degradation was detected by Western blotting. Effect of NF-B inhibitors, Bay11-7085 （10 μmol/L） and TPCK （20 μmol/L）, on Sin-mediated IL-8 expression in HGF, was detected by ELISA 24 h after pretreatment followed by 4 h-LPS stimulation. Results Sin suppressed the IL-8 mRNA and protein expressions in LPS-mediated HGF in dose-dependent manner （ P 〈 O. 05 ）. SIN could almost keep the NF-KB in the cytoplasm compared with LPS. Most NF-KB was transported in the nuclei 1 h after LPS treatment. The NF-KB level was 2-fold higher than the LPS level in cytoplasm after Sin （100 μmol/L） pretreatment. Fifty percent of IKBa was degraded 1 h after the HGF were pretreated with LPS and Sin inhibited the LPS-mediated degradation of IKB in a dose-dependent manner after HGF were pretreated （P 〈 0.05 ）. Sin （50 μmol/L） pretreatment returned the LPS-mediated IKBa degradation to its normal levels. The IL-8 expression level was 2.4-fold higher in LPS pretreatment group than in control group, and 1.8-fold and 1.6-fold lower in LPS pretreatment group than in control group after pretreatment with NF-KB inhibitors, Bayll-7085 （10 μmol/L） and TPCK （20 μmol/L）. Conclusion Sin inhibits the porphyromonasgingivalis LPS-mediated IL-8 expression in HGF by blocking the NF-KB translocation and the IKBct degradation.

关 键 词：青藤碱 牙龈 成纤维细胞 白细胞介素-8 NF-κB 

分 类 号：R285.5[医药卫生—中药学] R392.3[医药卫生—中医学]