中华皮肤科杂志
中華皮膚科雜誌
중화피부과잡지
Chinese Journal of Dermatology
2011年
7期
501-504
,共4页
郭英军%肖汀%王雅坤%陈洪铎%赵玉铭
郭英軍%肖汀%王雅坤%陳洪鐸%趙玉銘
곽영군%초정%왕아곤%진홍탁%조옥명
角蛋白细胞%抗原,CD68%细胞因子类%脂多糖类
角蛋白細胞%抗原,CD68%細胞因子類%脂多糖類
각단백세포%항원,CD68%세포인자류%지다당류
Keratinocytes%Antigens,CD68%Cytokines%Lipopolysaccharides
目的 研究肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)和干扰素γ(IFN-γ)以及细菌脂多糖(LPS)对CD68抗原在角质形成细胞株HaCaT细胞中表达的影响.方法 RPMI 1640培养液培养的HaCaT细胞随机分成自然增殖孔(无刺激组)、IFN-γ刺激孔(组)、TNF-α刺激孔(组)、INS刺激孔(组)、IL-6刺激孔(组).培养24 h后收集细胞,采用流式细胞仪、免疫组化和RT-PCR方法检测TNF-α、IL-6、IFN-γ和LPS作用HaCaT细胞后CD68表达情况.结果 与无刺激组比较,不同刺激因素可使HaCaT细胞CD68阳性细胞数增多,但以TNF-α和IL-6的作用较强(t值为3.60和3.93,P值均<0.01),IFN-γ和LPs的作用稍弱(t值为2.38和2.52,P值均<0.05).只有IL-6可以使HaCaT细胞CD68阳性细胞平均荧光强度增强(t值为8.34,P值<0.01 o在IFN-γ和TNF-α、IL-6作用HacaT细胞24 h后均有CD68表达于胞质和胞膜,并且以TNF-α和IL-6作用较强.TNF-α和IL-6作用HaCaT细胞24 h后可见CD68mRNA表达明显增强(t值为4.34和7.52,P值均<0.01),IFN-γ作用后出现较弱表达(t=2.81,P<0.05).结论 在IL-6、TNF-α、IFN-γ和LPS的作用下HaCaT细胞CD68表达增强.
目的 研究腫瘤壞死因子-α(TNF-α)、白介素-6(IL-6)和榦擾素γ(IFN-γ)以及細菌脂多糖(LPS)對CD68抗原在角質形成細胞株HaCaT細胞中錶達的影響.方法 RPMI 1640培養液培養的HaCaT細胞隨機分成自然增殖孔(無刺激組)、IFN-γ刺激孔(組)、TNF-α刺激孔(組)、INS刺激孔(組)、IL-6刺激孔(組).培養24 h後收集細胞,採用流式細胞儀、免疫組化和RT-PCR方法檢測TNF-α、IL-6、IFN-γ和LPS作用HaCaT細胞後CD68錶達情況.結果 與無刺激組比較,不同刺激因素可使HaCaT細胞CD68暘性細胞數增多,但以TNF-α和IL-6的作用較彊(t值為3.60和3.93,P值均<0.01),IFN-γ和LPs的作用稍弱(t值為2.38和2.52,P值均<0.05).隻有IL-6可以使HaCaT細胞CD68暘性細胞平均熒光彊度增彊(t值為8.34,P值<0.01 o在IFN-γ和TNF-α、IL-6作用HacaT細胞24 h後均有CD68錶達于胞質和胞膜,併且以TNF-α和IL-6作用較彊.TNF-α和IL-6作用HaCaT細胞24 h後可見CD68mRNA錶達明顯增彊(t值為4.34和7.52,P值均<0.01),IFN-γ作用後齣現較弱錶達(t=2.81,P<0.05).結論 在IL-6、TNF-α、IFN-γ和LPS的作用下HaCaT細胞CD68錶達增彊.
목적 연구종류배사인자-α(TNF-α)、백개소-6(IL-6)화간우소γ(IFN-γ)이급세균지다당(LPS)대CD68항원재각질형성세포주HaCaT세포중표체적영향.방법 RPMI 1640배양액배양적HaCaT세포수궤분성자연증식공(무자격조)、IFN-γ자격공(조)、TNF-α자격공(조)、INS자격공(조)、IL-6자격공(조).배양24 h후수집세포,채용류식세포의、면역조화화RT-PCR방법검측TNF-α、IL-6、IFN-γ화LPS작용HaCaT세포후CD68표체정황.결과 여무자격조비교,불동자격인소가사HaCaT세포CD68양성세포수증다,단이TNF-α화IL-6적작용교강(t치위3.60화3.93,P치균<0.01),IFN-γ화LPs적작용초약(t치위2.38화2.52,P치균<0.05).지유IL-6가이사HaCaT세포CD68양성세포평균형광강도증강(t치위8.34,P치<0.01 o재IFN-γ화TNF-α、IL-6작용HacaT세포24 h후균유CD68표체우포질화포막,병차이TNF-α화IL-6작용교강.TNF-α화IL-6작용HaCaT세포24 h후가견CD68mRNA표체명현증강(t치위4.34화7.52,P치균<0.01),IFN-γ작용후출현교약표체(t=2.81,P<0.05).결론 재IL-6、TNF-α、IFN-γ화LPS적작용하HaCaT세포CD68표체증강.
Objecfive To study the effects of some cytokines such as TNF-α,IL-6 and IFN-γ as well as lipopolysaccharide on CD68 expression in HaCaT cells.Methods Human HaCaT keratinocytes were randomly divided into natural proliferation group (without stimulation),IFN-γ-stimulated group,TNF-α-stimulated group,LPS-stimulated group and IL-6 stimulated group.The work concentration of TNF-α,IL-6,IFN-γ and LPS was 50 mg/L.HaCaT cells were collected after 24-hour treatment with the cytokines followed by the examination of CD68 expression with flow cytometry,immunohistochemistry and reverse transcription(RT)-PCR,respectively.Results Compared with untreated HaCaT cells,the count of CD68-positive cells was elevated in cells stimulated by TNF-α(t=3.60,P<0.01),IL-6(t=3.93,P<0.01),IFN-γ(t=2.38,P<0.05)and LPS(t=2.52,P<0.05),and the effect of TNF-α and IL-6 was stronger than that of IFN-γ and LPS.Among the four cytokines,only IL-6 enhanced the mean fluorescence intensity of CD68-positive cells (t=8.34,P<0.01).After 24-hour treatment with TNF-α,IFN-γ and IL-6,CD68 expression was observed in the cytoplasm and on the membrane of HaCaT cells and was stronger in cells treated with TNF-α and IL-6 than in those with the other cytokines.A significant increase was observed in the CD68 mRNA expression after 24-hour treatment with TNF-α (t=4.34,P<0.01),IL-6 (t=7.52,P<0.01)and IFN-γ (t=2.81,P<0.05);TNF-α and IL-6showed a stronger promotive effect than IFN-γ.Conclusion IL-6,TNF-α,IFN-γ and LPS can upregulate the CD68 expression in HaCaT cells.
