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

摘要:

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。

Abstract:

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^-1could increase the proliferation (P<0.05). The concentration of 125 mg ·L^-1could 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.