杜仲对紫外线致ESF-1细胞光老化保护作用的研究

Protective Effect of Eucommia ulmoides on Photoaging Induced by Ultraviolet Radiation in ESF-1 Cell

目的:研究杜仲抗紫外线致ESF-1细胞光老化作用,并初步探讨其作用机制。方法:杜仲70%乙醇提取物上D101大孔树脂制备不同浓度乙醇梯度洗脱部位,采用40 J.cm-2的UVA或70 mJ.cm-2的UVB照射ESF-1细胞后,立即加入不同浓度的杜仲50%乙醇洗脱部位,24 h后MTT法测定细胞活性,试剂盒法测定细胞培养液中乳酸脱氢酶(LDH)、超氧化歧化酶(SOD)活力及丙二醛(MDA)含量。结果:杜仲50%乙醇大孔树脂洗脱部位250,500 mg.L-1的给药浓度能使UVA诱导的光老化细胞活性明显增强(P<0.05),细胞培养液中LDH活力明显降低(P<0.05),其中250 mg.L-1的给药浓度能使细胞培养液中SOD活力明显升高(P<0.05),MDA含量明显降低(P<0.05);125,250,500 mg.L-1的给药浓度能使UVB诱导的光老化细胞活性明显增强,其中125 mg.L-1的给药浓度能使细胞培养液中LDH活力明显降低(P<0.05),SOD活力明显升高(P<0.05),MDA含量明显降低(P<0.05)。结论:杜仲50%乙醇大孔树脂洗脱部位对UVA和UVB致ESF-1细胞光老化具有良好的保护作用,推测其机制为通过提高SOD活力、加速氧自由基的清除和减少氧自由基的产生,使细胞的脂质过氧化损伤程度降低;同时,可能通过抗氧化作用,维持了细胞膜结构和功能的完整性,使LDH的漏出减少。

Objective：To study the protective effect of Eucommia ulmoides agains UV irradiation-induced damage in ESF-1 cells and its mechanism.Method： E.ulmoides was extracted by 70 ethanol,then the extract was eluted with gradient of ethanol by macroporous resin,ESF-1 cells were immediately treated with different concentrations of 50% ethanol eluate after UVA irradiation（40 J.cm-2） or UVB irradiation（70 mJ.cm-2）.Twenty-four hours later,the proliferation of cells was detected by MTT,SOD,LDH and MDA content were assayed by colrorimetry and TBA methods respectively.Result： Compared with the UVA group,the concentration of 125,250 mg.L-1 could increase the proliferation（P 0.05） and reduce the activity of LDH（P 0.05）.The concentration of 250 mg.L-1 could increase the activitiy of SOD（P 0.05） and decreased the content of MDA（P 0.05）.Compared with the UVB group,the concentration of 125,250 and 500 mg.L-1 could increase the proliferation（P 0.05）.The concentration of 125 mg.L-1 could increase the activity of SOD（P 0.05） and reduce the activity of LDH（P 0.05） and decreased the content of MDA（P 0.05）.Conclusion： The ethanol eluate of 50% can inhibit UV irradiation-induced damage in ESF-1 cells.Its mechanism is due to its abilities of increasing SOD,scavenging oxygen free radicals,inhibiting lipid peroxidation,and protecting membrane structure.