枸杞多糖对紫外线致人皮肤成纤维细胞脂质损伤的防御作用  

Effect of Lycium Barbarum Polysaccharide on UV-mediated Lipid Peroxide Damage of HSF Cells

作　　者：李振洁[1] 彭丽倩[1,2] 高爱莉[1] 张尔婷 刘清[1] 江娜[1] 李华平[1] 梁碧华[1] 李润祥[1] 代歆悦 莫子茵 朱慧兰[1] LI Zhen-jie PENG Li-qian GAO Ai-li ZHANG Er-ting LIU Qing JIANG Na LI Hua-ping LIANG Bi-hua LI Run-xiang DAI Xin-Yue~ MO Zi-yin ZHU Hui-lan(Guangzhou Institute of Dermatology, Guangzhou 510095, China Guangzhou Medical University)

机构地区：[1]广州市皮肤病防治所皮肤科,广州市5110095 [2]广州医科大学

出　　处：《中国激光医学杂志》2017年第3期127-132,共6页Chinese Journal of Laser Medicine & Surgery

基　　金：广东省建设中医药强省科研课题(20141221);2014年广东省医学科研基金(A2014592);2015年广州市科技计划项目(1563000503);2015年广东省自然科学基金项目(2015A030313765);2016年广州科技计划项目(201604020093)

摘　　要：目的探讨枸杞多糖(Lycium barbarum polysaccharide,LBP)对紫外线(Ultraviolet.,UV)照射的人皮肤成纤维细胞(Human Skin Fibroblasts,HSF)脂质损伤的防御作用及其机制。方法采用不同浓度LBP处理体外培养HSF细胞,用噻唑蓝(Methyl Thiazolyl Tetrazolium,MTT)法筛选出适宜药物浓度,行长波紫外线(Ultraviolet A,UVA)、中波紫外线(Ultraviolet B,UVB)照射建立HSF细胞急性光损伤模型,设空白组、LBP组、UVA组、UVA+LBP组、UVB组、UVB+LBP组;尼罗红荧光染色检测过氧化脂质含量;酶生化法测定胞质超氧化物歧化酶(Superoxide Dismutase,SOD)、谷胱甘肽过氧化物酶(Glutathione peroxidase,GSH-Px)活性及乳酸脱氢酶(Lactate dehydrogenase,LDH)漏出量。结果 LBP浓度在300μg/ml以下时,HSF细胞的增殖活性不受影响(P>0.05),当浓度>300μg/ml时,LBP抑制细胞增殖能力(P<0.05);与空白组比较,UVA和UVB照射组氧化脂质含量、LDH漏出量明显升高,胞内SOD、GSH-Px活性显著降低,差异均有统计学意义(均P<0.05);相比UV组,光照前加入LBP(UVA+LBP组、UVB+LBP组)可明显提高细胞SOD、GSH-Px活性,降低ROS水平、过氧化脂质含量及LDH漏出率(均P<0.05)。结论枸杞多糖可有效拮抗UV诱导的HSF细胞脂质损伤,作用机制可能与其增强SOD、GSH-Px等抗氧化酶的活性,降低LDH漏出量,保护细胞膜免受损伤有关。Objective To study the effect of Lycium barbarum polysaccharide on lipid peroxide damage of Human Skin Fibroblasts （HSF） due to irradiation of UV and its mechanism. Methods HSF cells were cultured in vitro to establish acute photo damage models with irradiation of UV. For an appropriate drug con- centration, MTT colorimetry was used to detect the viability of HSF cells. The cells were randomly divided into four groups： the blank control group, LBP group, UV radiated groups and UV ＋ LBP groups. Nile red staining and fluorescence microscopy observation was performed to determine the level of lipid peroxide. Enzyme biochemical methods were applied to measure the cytoplasmic SOD, GSH- Px activity and LDH leakage volume. Results When the concentration of LBP was below 300 μg/ml, the proliferative activity of HSF cells was unaffected （P 〉 0. 05 ）. However, LBP inhibited the cell proliferating when it was larger than 300 μg/ml （P 〈 0. 05）. Both the content of lipid peroxide and LDH ｜eakage of either the UVA or UVB group were significantly higher than those of the blank group. In addition, the content of SOD and GSH-Px dropped significantly （P 〈 0. 05 ）. Compared with other UV groups, LBP improved the activity of SOD and GSH-Px and decreased the amounts of lipid peroxide and LDH in the LBP groups, and the difference was also significant （P 〈 0. 05）. Conclusions LBP will effectively attenuate lipid oxidation damage of HSF ceils induced by UV, and its mechanism may be related to enhancing the activity of antioxidases, reducing the LDH leakage and protecting the cell membrane from injury.

关 键 词：枸杞 多糖 人皮肤成纤维细胞 紫外线 过氧化脂质 尼罗红 

分 类 号：R751[医药卫生—皮肤病学与性病学]