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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 photoaging effect of Eucommia ulmoides-induced UV-induced ESF-1 cells and to explore its mechanism. METHODS: Eucalyptus 70% ethanol extract was used to prepare gradient elution fractions of D101 macroporous resin with different concentration of ethanol. After ESF-1 cells were irradiated with UVA at 40 J.cm-2 or UVB at 70 mJ.cm-2, Concentration of eucommia bark 50% ethanol, cell viability was measured by MTT method 24 h later. The activities of lactate dehydrogenase (LDH), superoxide dismutase (SOD) and malondialdehyde (MDA) content. Results: The concentration of 250, 500 mg.L-1 of eucommia macroporous resin elution site could significantly increase the activity of UVA-induced photoaging cells (P <0.05) and the activity of LDH in cell culture medium significantly decreased (P < 0.05). The concentration of 250 mg.L-1 could significantly increase the activity of SOD in the cell culture medium (P <0.05) and decrease the content of MDA (P <0.05), while the dosage of 125 mg.L-1 The concentration of 125 mg.L-1 could significantly decrease the activity of LDH in the cell culture medium (P <0.05) and increase the activity of SOD (P <0.05) , MDA content decreased significantly (P <0.05). CONCLUSION: The eupeptide 50% ethanol macroporous elution site has a good protective effect on the photoaging of ESF-1 cells induced by UVA and UVB. It is presumed that the mechanism of EC50 is to improve the activity of SOD, accelerate the removal of oxygen free radicals and reduce the content of oxygen free radicals Resulting in decreased lipid peroxidation of cells; the same time, may be through the antioxidant effect, maintaining the integrity of the cell membrane structure and function, so that leakage of LDH reduced.