CAJ | 학술논문

目的探讨热处理对体外培养的正常人黑素细胞的增殖活性、黑素合成及其酪氨酸酶活性的影响.方法无菌操作获取正常人包皮环切术后的包皮,按照皮肤表皮细胞培养法获取黑素细胞,利用碱性成纤维细胞生长因子(bFGF)为主要有丝分裂原建立正常人表皮黑素细胞培养系,采用Masson-Fontana染色法对培养细胞进行鉴定,确定细胞种系,噻唑蓝(MTT)比色法测定不同温度条件(39、41、42、43、45 ℃)处理对黑素细胞活力的影响并选择最佳的温度条件,以左旋多巴为底物测定酪氨酸酶活性,比色法测定黑素含量,并将实验组与对照组结果进行比较.结果体外培养的正常人黑素细胞在5个不同的温度条件作用后,以42 ℃作用后的细胞增殖活性最佳.酪氨酸酶活性和黑素含量测定结果显示,42℃每天1 h连续干预3 d后,体外培养的正常人黑素细胞的酪氨酸酶活性增长率为36.4%,黑素合成增长率为78%,均明显高于对照组(P值均＜0.01).结论热处理可以增强体外培养的人黑素细胞的增殖活性,促进其黑素合成,并且可提高其酪氨酸酶的活性.
목적 탐토열처리대체외배양적정상인흑소세포적증식활성、흑소합성급기락안산매활성적영향.방법 무균조작획취정상인포피배절술후적포피,안조피부표피세포배양법획취흑소세포,이용감성성섬유세포생장인자(bFGF)위주요유사분렬원건립정상인표피흑소세포배양계,채용Masson-Fontana염색법대배양세포진행감정,학정세포충계,새서람(MTT)비색법측정불동온도조건(39、41、42、43、45 ℃)처리대흑소세포활력적영향병선택최가적온도조건,이좌선다파위저물측정락안산매활성,비색법측정흑소함량,병장실험조여대조조결과진행비교.결과 체외배양적정상인흑소세포재5개불동적온도조건작용후,이42 ℃작용후적세포증식활성최가.락안산매활성화흑소함량측정결과현시,42℃매천1 h련속간예3 d후,체외배양적정상인흑소세포적락안산매활성증장솔위36.4%,흑소합성증장솔위78%,균명현고우대조조(P치균＜0.01).결론 열처리가이증강체외배양적인흑소세포적증식활성,촉진기흑소합성,병차가제고기락안산매적활성.
Objective To investigate the effect of heat treatment on the proliferation of, melanin synthesis and tyrosinase activity in cultured normal human melanocytes. Methods Normal human foreskin tissue was obtained by sterile circumcision and melanocytes were harvested by using methods for epidermal cell culture. Basic fibroblast growth factor (bFGF) was utilized as the primary mitogen to establish the culture system of normal human epidermal melanocytes. Masson-Fontana staining was proformed to identify melanocytes.Third-passage melanocytes were treated with hyperthermia at various temperatures (39 ℃, 41 ℃, 42 ℃, 43 ℃ and 45℃) for 1 hour a day for consecutive 3 days followed by the measurement of cell viability with MTT assay. The hyperthermia at optimized temperature was used to treat fourth-passage melanocytes for 1 hour a day for consecutive 3 days; subsequently, the tyrosinase activity were detected with L-Dopa as the substrate, and melanin content was determined in heat-treated and untreated (control) melanocytes. Results The hyperthermia at 42 ℃ exhibited the strongest promotive effect on the proliferation of melanocytes among these 5 hyperthermia conditions. After treatment with hyperthermia at 42 ℃ for 1 hour a day for consecutive 3 days, melanocytes showed an increment in tyrosinase activity by 36.4% and melanin synthesis by 78% compared with the untreated melanocytes (both P＜0.05). Conclusions Heat treatment can enhance the proliferation of cultured human melanocytes, promote their melanin synthesis and tyrosinase activity.