中华皮肤科杂志
中華皮膚科雜誌
중화피부과잡지
Chinese Journal of Dermatology
2014年
12期
856-859
,共4页
杨子良%骆丹%钱齐宏%杜纳%余秀琴%王淼淼%闵玮
楊子良%駱丹%錢齊宏%杜納%餘秀琴%王淼淼%閔瑋
양자량%락단%전제굉%두납%여수금%왕묘묘%민위
黄芪%紫外线%角蛋白细胞%活性氧
黃芪%紫外線%角蛋白細胞%活性氧
황기%자외선%각단백세포%활성양
Astragalus membranaceus%Ultraviolet rays%Keratinocytes%Reactive oxygen species
目的 观察黄芪甲苷对于中波紫外线(UVB)损伤的人表皮细胞的保护作用,探讨其相关机制.方法 将培养的永生化人皮肤角质形成细胞(HaCaT细胞)分为对照组、UVB组、黄芪甲苷组和UVB+黄芪甲苷组,其中UVB组和UVB+黄芪甲苷组细胞接受50 mJ/cm2 UVB照射,加药组加入不同浓度的黄芪甲苷(10、20、50、100、200 mg/L)进行干预,24 h后CCK8法检测细胞活性.根据CCK8法检测结果选择最佳药物浓度(20 mg/L)进行后继实验.照光后继续培养24 h,流式细胞仪检测细胞内活性氧(ROS)水平,Western印迹法检测HaCaT细胞中p53、p38、基质金属蛋白酶9(MMP-9)和高迁移率族蛋白A1(HMGA-1)的表达.结果 与对照组相比,10 mg/L和20 mg/L黄芪甲苷组对HaCaT细胞的增殖活性无明显影响(F=1.32,P>0.05),50、100和200 mg/L黄芪甲苷对细胞增殖活性有一定抑制作用(F=20.20,P< 0.05);UVB组与对照组比较,HaCaT细胞增殖活性显著下降(F=99.00,P< 0.01).与UVB组相比,UVB+黄芪甲苷(10~ 200 mg/L)组HaCaT细胞增殖活性不同程度升高(F=19.08,P< 0.01),其中UVB+ 20 mg/L黄芪甲苷组升高程度最高.进一步实验表明,与UVB组相比,UVB+ 20 mg/L黄芪甲苷组ROS产生受到明显抑制(f=21.12,P< 0.01).Western印迹结果表明,与对照组比较,UVB组p53、p38、MMP-9和HMGA-1蛋白的表达升高(均P< 0.01),而UVB+ 20 m/L黄芪甲苷组细胞内p53、p38、MMP-9和HMGA-1蛋白的表达水平较UVB组显著降低(P<0.01).结论 黄芪甲苷可有效抑制UVB引起的表皮细胞光损伤.
目的 觀察黃芪甲苷對于中波紫外線(UVB)損傷的人錶皮細胞的保護作用,探討其相關機製.方法 將培養的永生化人皮膚角質形成細胞(HaCaT細胞)分為對照組、UVB組、黃芪甲苷組和UVB+黃芪甲苷組,其中UVB組和UVB+黃芪甲苷組細胞接受50 mJ/cm2 UVB照射,加藥組加入不同濃度的黃芪甲苷(10、20、50、100、200 mg/L)進行榦預,24 h後CCK8法檢測細胞活性.根據CCK8法檢測結果選擇最佳藥物濃度(20 mg/L)進行後繼實驗.照光後繼續培養24 h,流式細胞儀檢測細胞內活性氧(ROS)水平,Western印跡法檢測HaCaT細胞中p53、p38、基質金屬蛋白酶9(MMP-9)和高遷移率族蛋白A1(HMGA-1)的錶達.結果 與對照組相比,10 mg/L和20 mg/L黃芪甲苷組對HaCaT細胞的增殖活性無明顯影響(F=1.32,P>0.05),50、100和200 mg/L黃芪甲苷對細胞增殖活性有一定抑製作用(F=20.20,P< 0.05);UVB組與對照組比較,HaCaT細胞增殖活性顯著下降(F=99.00,P< 0.01).與UVB組相比,UVB+黃芪甲苷(10~ 200 mg/L)組HaCaT細胞增殖活性不同程度升高(F=19.08,P< 0.01),其中UVB+ 20 mg/L黃芪甲苷組升高程度最高.進一步實驗錶明,與UVB組相比,UVB+ 20 mg/L黃芪甲苷組ROS產生受到明顯抑製(f=21.12,P< 0.01).Western印跡結果錶明,與對照組比較,UVB組p53、p38、MMP-9和HMGA-1蛋白的錶達升高(均P< 0.01),而UVB+ 20 m/L黃芪甲苷組細胞內p53、p38、MMP-9和HMGA-1蛋白的錶達水平較UVB組顯著降低(P<0.01).結論 黃芪甲苷可有效抑製UVB引起的錶皮細胞光損傷.
목적 관찰황기갑감대우중파자외선(UVB)손상적인표피세포적보호작용,탐토기상관궤제.방법 장배양적영생화인피부각질형성세포(HaCaT세포)분위대조조、UVB조、황기갑감조화UVB+황기갑감조,기중UVB조화UVB+황기갑감조세포접수50 mJ/cm2 UVB조사,가약조가입불동농도적황기갑감(10、20、50、100、200 mg/L)진행간예,24 h후CCK8법검측세포활성.근거CCK8법검측결과선택최가약물농도(20 mg/L)진행후계실험.조광후계속배양24 h,류식세포의검측세포내활성양(ROS)수평,Western인적법검측HaCaT세포중p53、p38、기질금속단백매9(MMP-9)화고천이솔족단백A1(HMGA-1)적표체.결과 여대조조상비,10 mg/L화20 mg/L황기갑감조대HaCaT세포적증식활성무명현영향(F=1.32,P>0.05),50、100화200 mg/L황기갑감대세포증식활성유일정억제작용(F=20.20,P< 0.05);UVB조여대조조비교,HaCaT세포증식활성현저하강(F=99.00,P< 0.01).여UVB조상비,UVB+황기갑감(10~ 200 mg/L)조HaCaT세포증식활성불동정도승고(F=19.08,P< 0.01),기중UVB+ 20 mg/L황기갑감조승고정도최고.진일보실험표명,여UVB조상비,UVB+ 20 mg/L황기갑감조ROS산생수도명현억제(f=21.12,P< 0.01).Western인적결과표명,여대조조비교,UVB조p53、p38、MMP-9화HMGA-1단백적표체승고(균P< 0.01),이UVB+ 20 m/L황기갑감조세포내p53、p38、MMP-9화HMGA-1단백적표체수평교UVB조현저강저(P<0.01).결론 황기갑감가유효억제UVB인기적표피세포광손상.
Objective To evaluate the protective effect of astragaloside Ⅳ against ultraviolet B (UVB)-induced photodamage to human HaCaT keratinocytes,and to investigate its mechanisms.Methods Culturedimmortalized human HaCaT keratinocytes were divided into four groups:blank control group receiving untreated,UVB group irradiated with 50 mJ/cm2 UVB,astragaloside Ⅳ group treated with astragaloside Ⅳ,UVB + astragalosideⅣ group treated with astragaloside Ⅳ for 24 hours before and after 50 mJ/cm2 of UVB radiation.The concentration ofastragaloside Ⅳ ranged from 10 to 200 mg/L in cell proliferation assay,and according to the results of proliferationassay,20 mg/L was determined as the optimal concentration in the other assays.At 24 hours after UVB radiation,cellcounting kit-8 (CCK8) assay was performed to evaluate cellular proliferative activity,flow cytometry to determineintracellular reactive oxygen species (ROS) levels,and Western blot to measure the expression levels of p53,p38,matrix metalloproteinase-9 (MMP-9) and high mobility group Al (HMGA-1) protein in HaCaT cells.ResultsCompared with the control group,astragaloside Ⅳ at 10 and 20 mg/L had no inhibitory effect (F =1.32,P > 0.05),while astragaloside Ⅳ at 50,100 and 200 mg/L showed significantly inhibitory effect (F =20.20,P < 0.05),on theproliferation of HaCaT cells.In addition,cellular proliferative activity in the UVB group was significantly lower thanthat in the control group (F =99.00,P < 0.01).Compared with the UVB group,cellular proliferative activityincreased to different degrees in HaCaT cells treated with both UVB and astragaloside Ⅳ of 10-200 mg/L (F =19.08,P < 0.01),with the strongest increase observed in those treated with UVB and astragaloside Ⅳ of 20 mg/L.Further experiments revealed reduced intracellular ROS levels in the UVB + astragaloside Ⅳ (20 mg/L) groupcompared with the UVB group (t =21.12,P < 0.01).Western blot assay showed that the expression levels of p53,p38,MMP-9 and HMGA-1 protein were significantly higher in the UVB group than in the control group (all P <0.01),but significantly lower in the UVB + astragaloside Ⅳ (20 mg/L) group than in the UVB group (all P < 0.01).Conclusion Astragaloside Ⅳ can effectively protect keratinocytes from UVB-induced photodamage.