TLR2在皮肤癣菌感染中对角质形成细胞分泌γ-IFN和IL-8的影响

发布时间：2018-04-03 15:52

  本文选题：角质形成细胞　切入点：细胞培养　出处：《华中科技大学》2011年硕士论文

【摘要】：第一部分人角质形成细胞的原代培养 目的 建立人表皮角质形成细胞的分离和培养方法,为后续实验提供基础。 方法 取青少年包皮环切术后的包皮组织,经无菌处理后,用常用的胰酶消化法分离表皮和真皮,分离出的表皮细胞用无血清角质形成细胞培养基培养并用免疫细胞化学方法进行鉴定。 结果 用无血清培养基培养的角质形成细胞,成纤维细胞污染少,贴壁率高,荧光显微镜下细胞胞浆呈角蛋白阳性染色。6代以内的细胞活性均较好。 结论 用常规的酶消化法分离表皮,以无血清角质形成细胞培养基培养能成功培养出角质形成细胞。 第二部分TLR2在抗红色毛癣菌感染中对角质形成细胞分泌γ-IFN和IL-8的影响 目的 观察红色毛癣菌刺激人角质形成细胞后γ-IFN及IL-8的变化,以及TLR-2对γ-IFN和IL-8的分泌的影响;探讨TLR-2在抗皮肤癣菌感染中的作用。 方法 用红色毛癣菌悬液分别刺激TLR2抗体处理前后的角质形成细胞,采用ELISA方法检测不同时间点细胞上清液中γ-IFN及IL-8的浓度,并设置阴性对照;比较TLR2抗体处理前后γ-IFN及IL-8浓度的变化。 结果 红色毛癣菌刺激角质形成细胞后,γ-IFN及IL-8浓度明显升高(P�d0.05),γ-IFN在4h即达到(85.36±4.54)pg/ml,16h后达到(445.58±13.99)pg/ml ;IL-8在4h即达到(545.426±39.784)pg/ml,16h后达到(977.182±55.288)pg/ml;用TLR2抗体中和TLR2后,清液中IL-8的浓度在2h、4h、8h、16h各时间点较中和前低,差异有统计学意义(P�d0.05);γ-IFN的浓度2h、4h、8h时间点较中和前低,差异有统计学意义(P�d0.05),而在16h时间点,上清液中γ-IFN的浓度与中和前比较略低,但差异没有统计学意义(P�e0.05)。 结论 红色毛癣菌刺激角质形成细胞后,可促进角质形成细胞分泌γ-IFN和IL-8;TLR2在角质形成细胞分泌γ-IFN和IL-8的过程中发挥重要的调节作用。
[Abstract]:Part one primary culture of human keratinocytesPurposeThe method of isolation and culture of human epidermal keratinocytes was established to provide the basis for further experiments.MethodThe tissue of prepuce after circumcision of juvenile prepuce was removed. After sterile treatment, the epidermis and dermis were separated by common trypsin digestion method.The isolated epidermal cells were cultured on serum-free keratinocytes medium and identified by immunocytochemistry.ResultThe keratinocytes cultured in serum-free medium had less contamination and high adhesion rate. The cytoplasm of keratin positive staining could be found in the cytoplasm of the keratin positive cells under fluorescence microscope.ConclusionKeratinocytes were isolated by enzyme digestion and cultured on serum-free keratinocytes medium.The second part: the effect of TLR2 on the secretion of 纬 -IFN and IL-8 by keratinocytes against Trichophyton rubrum infectionPurposeTo observe the changes of 纬 -IFN and IL-8 in human keratinocytes stimulated by Trichophyton rubrum and the effect of TLR-2 on the secretion of 纬 -IFN and IL-8, and to explore the role of TLR-2 in anti-tinea dermatophytes infection.MethodKeratinocytes were stimulated by Trichophyton rubrum suspension before and after TLR2 antibody treatment. The concentrations of 纬 -IFN and IL-8 in supernatant of cells at different time points were detected by ELISA method, and negative control was set.To compare the changes of 纬 -IFN and IL-8 before and after TLR2 antibody treatment.Result绾㈣壊姣涚櫍鑿屽埡婵，

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