目的:探讨姜黄素对IL-17诱导的人表皮角质形成细胞株(HaCaT细胞)NO合成以及诱导型一氧化氮合酶(iN-OS)的mRNA和蛋白表达的影响。方法:用IL-17刺激体外培养的HaCaT细胞,并分别加入3种浓度的姜黄素共培养24 h。并收集细胞上清液、提取细胞总RNA、总蛋白,分别进行NO含量的测定、荧光定量PCR和W estern b lot实验,明确姜黄素对NO含量以及iNOS表达的影响。结果:IL-17能够有诱导HaCaT细胞NO以及iNOS的表达(P<0.01)。姜黄素有效下调NO合成量以及iNOS的mRNA(P<0.01)及蛋白表达(P<0.01)水平。结论:姜黄素对IL-17诱导的HaCaT细胞NO分泌及iNOS的表达具有明显的抑制作用,从而为其对角质形成细胞相关的皮肤炎症性疾病的治疗提供了理论依据。
Objective: To study apoptotic effects of synthetic retinoic acid 6-[3-(1-adamantyl)-4-hydroxyphenyl]-2-naphthalene carboxylic acid(AHPN) on human skin malignant melanoma A375 cells in comparison with the natural ligand all-trans-retinoic acid(ATRA) in vitro and the mechanisms related to the actions of AHPN. Methods:MTT assay was used to determine the anti-proliferative effects of AHPN and ATRA on A375 cells. Flow cytometry was performed to investigate the influence of AHPN and ATRA on cell cycle and cell apoptosis. In addition, transfection and luciferase activity assays were employed to explore the mechanisms of how AHPN executes its proapoptotic function. Results:Firstly, AHPN promoted apoptosis and GI arrest in A375 cells compared with ATRA. Secondly, the activity of NF-K B in A375 cells treated with AHPN increased 2-3 times compared with solvent DMSO treatment. Conclusion:AHPN, in comparison with ATRA, is a more effective alternative for therapy of malignant melanoma. The potentially proapoptotic function of AHPN requires activation of NF-K B.
Min Pan Zhen-hui Peng Sheng-xiang Xiao Jian-wen Ren Yan Liu Xiao-li Li Zheng-xiao Li
Objective: To investigate the effects of synthetic retinoid CD437 and acitretin on cell proliferation, apoptosis, cycle arrest and Bax/ Bcl-2 protein expression of melanoma A375 cell, Methods:MTT assay was used to determine the anti-proliferative effects of CD437 and acitretin on melanoma A375 cell, Flow cytometry was performed to investigate the influence of CD437 and acitretin on cell cycle and cell apoptosis. SABC immunocytochemistry was employed for detection of Bax/bcl-2 protein expressions. Results:10^-5 mol/L CD437 was more effective than acitretin in inhibiting proliferation and inducing apoptosis of A375 cell after 24 h treatment, growth inhibiting ratio and apoptosis ratio(58.6%vs43.25% and 28.03%vs17.13%, P 〈 0.05 respectively). CD437 promoted G0/G1 arrest in melanoma A375 cell, however acitretin could not. CD437 and acitretin could up-regulate the expression of Bax protein and downregulate the expression of bcl-2 protein(P 〈 0.05). Conclusion:CD437 is more effective than acitretin in inhibiting proliferation and inducing apoptosis and cycle arrest on A375 cell, CD437 may have more potentialities than acitretin for subsidiary treatment of melanoma. Mitochondrial apoptosis pathway is partially involved in two drugs inducing apoptosis on A375 cell.
Jianwen Ren Zhenhui Peng Min Pan Birong Guo Yan Liu Xianglan Wang