中华皮肤科杂志
中華皮膚科雜誌
중화피부과잡지
Chinese Journal of Dermatology
2013年
11期
810-814
,共5页
陈凰%李远宏%徐学刚%吴严
陳凰%李遠宏%徐學剛%吳嚴
진황%리원굉%서학강%오엄
紫外线%成纤维细胞%细胞因子类%白藜芦醇
紫外線%成纖維細胞%細胞因子類%白藜蘆醇
자외선%성섬유세포%세포인자류%백려호순
Ultraviolet rays%Fibroblasts%Cytokines%Resveratrol
目的 探讨白藜芦醇对长波紫外线(UVA)照射的人成纤维细胞的保护作用及其机制.方法 0.01、0.1、0.5、1.0 mmol/L白藜芦醇分别处理正常人成纤维细胞6、24、48、72 h后,采用MTT法检测增殖活性.10 J/cm2 UVA照射细胞后立即加入0.01、0.1 mmol/L白藜芦醇,继续培养6、24、48、72 h,MTT法检测细胞的增殖活性,ELISA法检测上清液中白介素(IL)-1α、IL-1β、IL-6蛋白水平.结果 0.5、1.0 mmol/L白藜芦醇对正常人成纤维细胞增殖活性有明显抑制作用,72 h时抑制作用最为显著,其细胞存活率分别为3 1.99％±8.29％和21.15％±5.76％.单独UVA照射后6h,细胞存活率为78.01％±12.74％,直至照射后72 h细胞存活率(80.64％±36.12％)仍低于空白对照组(99.95％±12.23％)；照射后6h,培养上清液中IL-1α、IL-1β、IL-6蛋白水平分别升高至(58.39±0.67) ng/L、(1294.37±92.51) pg/L、(197.81±6.37) ng/L,直至照射后72 h,IL-1β、IL-6蛋白分泌量仍处于高水平(1236.76±56.49) pg/L、(215.65±3.78) ng/L.成纤维细胞经UVA照射加白藜芦醇后,与单独UVA照射组比较,0.01 mmol/L白藜芦醇组在各观察点均使细胞存活率升高,作用72 h时细胞存活率升高至91.93％±12.90％,并持续抑制细胞分泌的IL-1α蛋白至48 h(43.89±3.60 ng/L)、IL-1β蛋白至72 h(1110.12±51.91 pg/L)、IL-6蛋白至72 h(201.94±4.71 ng/L);而0.1 mmol/L白藜芦醇组在各观察点对细胞存活率均无明显影响,仅持续抑制IL-6蛋白水平至24 h(182.90±6.67 ng/L).结论 0.01 mmol/L白藜芦醇对UVA照射损伤的成纤维细胞有保护作用,其机制可能与白藜芦醇抑制UVA诱导的细胞因子IL-1α、IL-1β、IL-6分泌有关.
目的 探討白藜蘆醇對長波紫外線(UVA)照射的人成纖維細胞的保護作用及其機製.方法 0.01、0.1、0.5、1.0 mmol/L白藜蘆醇分彆處理正常人成纖維細胞6、24、48、72 h後,採用MTT法檢測增殖活性.10 J/cm2 UVA照射細胞後立即加入0.01、0.1 mmol/L白藜蘆醇,繼續培養6、24、48、72 h,MTT法檢測細胞的增殖活性,ELISA法檢測上清液中白介素(IL)-1α、IL-1β、IL-6蛋白水平.結果 0.5、1.0 mmol/L白藜蘆醇對正常人成纖維細胞增殖活性有明顯抑製作用,72 h時抑製作用最為顯著,其細胞存活率分彆為3 1.99％±8.29％和21.15％±5.76％.單獨UVA照射後6h,細胞存活率為78.01％±12.74％,直至照射後72 h細胞存活率(80.64％±36.12％)仍低于空白對照組(99.95％±12.23％)；照射後6h,培養上清液中IL-1α、IL-1β、IL-6蛋白水平分彆升高至(58.39±0.67) ng/L、(1294.37±92.51) pg/L、(197.81±6.37) ng/L,直至照射後72 h,IL-1β、IL-6蛋白分泌量仍處于高水平(1236.76±56.49) pg/L、(215.65±3.78) ng/L.成纖維細胞經UVA照射加白藜蘆醇後,與單獨UVA照射組比較,0.01 mmol/L白藜蘆醇組在各觀察點均使細胞存活率升高,作用72 h時細胞存活率升高至91.93％±12.90％,併持續抑製細胞分泌的IL-1α蛋白至48 h(43.89±3.60 ng/L)、IL-1β蛋白至72 h(1110.12±51.91 pg/L)、IL-6蛋白至72 h(201.94±4.71 ng/L);而0.1 mmol/L白藜蘆醇組在各觀察點對細胞存活率均無明顯影響,僅持續抑製IL-6蛋白水平至24 h(182.90±6.67 ng/L).結論 0.01 mmol/L白藜蘆醇對UVA照射損傷的成纖維細胞有保護作用,其機製可能與白藜蘆醇抑製UVA誘導的細胞因子IL-1α、IL-1β、IL-6分泌有關.
목적 탐토백려호순대장파자외선(UVA)조사적인성섬유세포적보호작용급기궤제.방법 0.01、0.1、0.5、1.0 mmol/L백려호순분별처리정상인성섬유세포6、24、48、72 h후,채용MTT법검측증식활성.10 J/cm2 UVA조사세포후립즉가입0.01、0.1 mmol/L백려호순,계속배양6、24、48、72 h,MTT법검측세포적증식활성,ELISA법검측상청액중백개소(IL)-1α、IL-1β、IL-6단백수평.결과 0.5、1.0 mmol/L백려호순대정상인성섬유세포증식활성유명현억제작용,72 h시억제작용최위현저,기세포존활솔분별위3 1.99％±8.29％화21.15％±5.76％.단독UVA조사후6h,세포존활솔위78.01％±12.74％,직지조사후72 h세포존활솔(80.64％±36.12％)잉저우공백대조조(99.95％±12.23％)；조사후6h,배양상청액중IL-1α、IL-1β、IL-6단백수평분별승고지(58.39±0.67) ng/L、(1294.37±92.51) pg/L、(197.81±6.37) ng/L,직지조사후72 h,IL-1β、IL-6단백분비량잉처우고수평(1236.76±56.49) pg/L、(215.65±3.78) ng/L.성섬유세포경UVA조사가백려호순후,여단독UVA조사조비교,0.01 mmol/L백려호순조재각관찰점균사세포존활솔승고,작용72 h시세포존활솔승고지91.93％±12.90％,병지속억제세포분비적IL-1α단백지48 h(43.89±3.60 ng/L)、IL-1β단백지72 h(1110.12±51.91 pg/L)、IL-6단백지72 h(201.94±4.71 ng/L);이0.1 mmol/L백려호순조재각관찰점대세포존활솔균무명현영향,부지속억제IL-6단백수평지24 h(182.90±6.67 ng/L).결론 0.01 mmol/L백려호순대UVA조사손상적성섬유세포유보호작용,기궤제가능여백려호순억제UVA유도적세포인자IL-1α、IL-1β、IL-6분비유관.
Objective To investigate the protective effect of resveratrol on ultraviolet A (UVA)-irradiated human fibroblasts and its mechanism.Methods Fibroblasts were isolated from normal human foreskin and subjected to primary culture and four passages of subculture.Then,some fibroblasts were incubated with various concentrations (0.01,0.1,0.5 and 1.0 mmol/L) of resveratrol for 6,24,48 and 72 hours separately,followed by methyl thiazolyl tetrazolium (MTT) assay for the evaluation of cell proliferation.Some fibroblasts were classified into four groups:blank control group remaining untreated,UVA group irradiated with UVA only,0.01 and 0.1 mmol/L resveratrol groups receiving UVA irradiation immediately followed by treatment with resveratrol of 0.01 and 0.1 mmol/L respectively.The dose of UVA irradiation was consistently 10 J/cm2 in these groups.After additional culture for 6,24,48 and 72 hours,MTT assay was conducted to evaluate cell proliferation,enzyme-linked immunosorbent assay (ELISA) to measure the levels of interleukin (IL)-1α,IL-1β,and IL-6 in the culture supernatant.Results Resveratrol at 0.5 and 1.0 mmol/L significantly inhibited the proliferation of fibroblasts,with the strongest inhibitory effect observed at 72 hours when the cell survival rate was 31.99％ ± 8.29％ and 21.15％ ± 5.76％,respectively.After irradiation with UVA of 10 J/cm2,the survival rate of fibroblasts was 78.01％ ± 12.74％ at 6 hours and 80.64％ ± 36.12％ at 72 hours,compared to 100.04％ ± 10.78％ and 99.95％ ± 12.23％ in the blank control group respectively (both P ＜ 0.05); the supernatant levels of IL-1α,IL-1β and IL-6 were significantly increased compared with the blank control group at 6 hours ((58.39 ± 0.67) vs.(48.51 ± 6.20) ng/L,(1294.37 ± 92.51) vs.(1023.25 ± 86.40) pg/L,(197.81 ± 6.37) vs.(160.45 ± 7.19) ng/L,all P ＜ 0.05),and the increase still existed at 72 hours for IL-1β ((1236.76 ± 56.49) vs.(1045.55 ± 48.14) pg/L,P＜ 0.05) and IL-6 ((215.65 ± 3.78) vs.(195.09 ± 1.78) ng/L,P ＜ 0.05).Compared with the UVA group,the 0.01 mmol/L resveratrol group showed significantly higher survival rates at all the four time points (all P ＜ 0.05),but lower supernatant levels of IL-1α at 6,24 and 48 ((43.89 ± 3.60) vs.(51.77 ± 1.77) ng/L,P＜ 0.05) hours as well as IL-lβ and IL-6 at all the four time points (all P ＜ 0.05),while the 0.1 mmol/L resveratrol group experienced no significant changes in cell survival rate at any of the time points,with a significant decrease only in the supernatant level of IL-6 at 6 and 24 ((182.90 ± 6.67) vs.(240.62 ± 1.42) ng/L,P ＜ 0.05) hours.In detail,the survival rate of fibroblasts was 91.93％ ± 12.90％,with the supernatant level being (1110.12 ± 51.91) pg/L for IL-1β and (201.94 ± 4.71) ng/L for IL-6 at 72 hours in the 0.01 mmol/L resveratrol group,compared to 80.64％ ± 36.12％,(1236.76 ± 56.49) pg/L and (215.65 ± 3.78) ng/L respectively in the UVA group (all P＜ 0.05).Conclusion Resveratrol at 0.01 mmol/L has a protective effect on UVA-irradiated fibroblasts,likely by inhibiting the secretion of IL-1α,IL-1β and IL-6.
