疗癣卡西甫散提取物对肿瘤坏死因子―α诱导角质形成细精品分析.doc

疗癣卡西甫散提取物对肿瘤坏死因子―α诱导角质形成细胞增殖的影响 　　摘要：目的 观察疗癣卡西甫散提取物（LE）对肿瘤坏死因子-α（TNF-α）诱导角质形成细胞株HaCaT细胞增殖及白细胞介素-8（IL-8）、细胞间黏附分子-1（ICAM-1）mRNA表达的影响，探讨其作用机制。方法 将培养的HaCaT细胞分为正常组、TNF-α组和LE低、中、高剂量组。除正常组外，其余各组细胞均给予40 ng/mL TNF-α诱导，各给药组再分别给予8、40、200 μg/mL LE作用48 h。MTT法检测HaCaT细胞增殖；ELISA检测HaCaT细胞IL-8和ICAM-1含量；PI和Hoechst33342荧光染色观察HaCaT细胞凋亡；半定量RT-PCR检测HaCaT细胞IL-8和ICAM-1的mRNA表达。结果 与正常组比较，TNF-α组HaCaT细胞吸光度、IL-8和ICAM-1含量及其mRNA表达明显升高（P0.05，P0.01）；与TNF-α组比较，LE各剂量组HaCaT细胞吸光度、IL-8和ICAM-1含量及其mRNA表达显著降低（P0.05，P0.01）。结论 LE通过促进HaCaT细胞凋亡及抑制HaCaT细胞IL-8和ICAM-1基因表达，从而维持皮损处HaCaT细胞的正常水平。 　　关键词：疗癣卡西甫散提取物；肿瘤坏死因子-α；HaCaT；细胞增殖；白细胞介素-8；细胞间黏附分子-1 　　DOI：10.3969/j.issn.1005-5304.2016.11.016 　　R285.5 A 1005-5304（2016）11-0067-04 　　Abstract： Objective To investigate the effects of extracts of Liaoxuan Kaxifu Powders （LE） on proliferation and transcription of IL-8 and ICAM-1 in HaCaT induced by TNF-α； To discuss its mechanism of action. Methods Cultured HaCaT was assigned into normal group， TNF-α group and low-， medium- and high-dose of LE group. Every group was induced by 40 ng/mL TNF-α except for normal group， and then LE groups were treated by different concentrations （8， 40， 200 μg/mL） of LE for 48 h. The proliferation of HaCaT was evaluated by MTT and the apoptosis was detected by inverted fluorescence microscope. Levels of IL-8 and ICAM-1 in HaCaT were assessed by ELISA， and their mRNA expressions was detected by semi-quantitative RT-PCR. Results Compared with normal group， the absorbency of HaCaT and the contents and mRNA expressions of IL-8 and ICAM-1 increased in TNF-α group （P0.05， P0.01）； compared with TNF-α group， LE of all dose groups could significantly inhibit the absorbency， decrease the contents and mRNA expressions of IL-8 and ICAM-1 （P0.05， P0.01）. Conclusion LE is able to inhibit the proliferation of HaCaT induced by TNF-α， and the mechanism is probably related to promoting apoptosis and down-regulating the gene expressions of IL-8 and ICAM-1， and then maintaining the normal