中国麻风皮肤病杂志 ›› 2018, Vol. 34 ›› Issue (10): 580-584.

• 论著 • 上一篇    下一篇

葡萄籽原花青素对中波紫外线诱导HaCaT细胞氧化损伤的保护作用

李桂双1 卜洁琼2 龙剑文3   

  1. 1.山东省日照市皮肤病防治所，日照，276800；2山东省日照市疾病预防控制中心，日照，276800；3.湖北中医药大学第一临床学院皮肤科，武汉，430061
  • 出版日期:2018-10-15 发布日期:2018-12-03
  • 通讯作者: 龙剑文，E-mail: longjianwen@hbhtcm.com

Protective effect of proanthocyanidin on ultraviolet B-induced oxidative damage to HaCaT cells

LI Guishuang1 ,BU Jieqiong2 ,LONG Jianwen3   

  1. 1.Rizhao Prevention & Cure Institute of Dermatopathy, Rizhao276800, China; 2. Rizhao Center for Disease Control and Prevention, Rizhao276800, China; 3. Department of Dermatology, the First Clinical Medical School, Hubei University of Chinese Medicine, Wuhan 430061, China
  • Online:2018-10-15 Published:2018-12-03
  • Contact: LONG Jianwen, E-mail: longjianwen@hbhtcm.com

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摘要： 目的: 明确葡萄籽原花青素（ GSPE）对 UVB损伤角质形成细胞的保护作用。方法: 体外培养HaCaT细胞，分为正常对照组、UVB组、UVB+GSPE高剂量(200 μg/mL) 、中剂量(100 μg/mL)和低剂量组(50μg/mL)。CCK-8法检测HaCaT细胞增殖能力；Annexin V-FITC/PI染色后，流式细胞仪检测细胞凋亡率；比色法检测细胞超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性，丙二醛(MDA)和LDH水平。DCFH-DA染色法测定细胞内活性氧水平；罗丹明123染色法检测细胞线粒体膜电位水平。Western blot检测细胞内Bax、Bcl-2、Cleaved-caspase-3蛋白表达。结果: UVB+GSPE组与UVB组比较，细胞SOD和GSH-Px活性、Bcl-2 表达量和线粒体膜电位水平增高，MDA、LDH水平、凋亡细胞数、Bax、Cleaved-caspase-3表达和细胞内活性氧水平明显降低。结论: GSPE对中波紫外线辐射诱导的表皮角质形成细胞凋亡、氧化损伤具有一定的保护作用。

关键词: HaCaT细胞, 中波紫外线, 葡萄籽原花青素, 氧化损伤

Abstract: Objective: To determine the protective effect of grape seed proanthocyanidin extract (GSPE) on the oxidative stress of HaCaT cells induced by UVB. Methods: HaCaT cells were incubated in vitro and divided into control group, UVB group, UVB+GSPE groups (200 μg/mL, 100 μg/mL and 50 μg/mL). The viability of HaCaT cells was detected by CCK-8 assay. The apoptosis of HaCaT cells was examined by flow cytometry assay. The level of SOD, GSH-Px, MDA and LDH was detected by colorimetry．The level of reactive oxygen species (ROS) and mitochondrial membrane potential were evaluated by DCFH-DA staining and rhodamine 123 staining respectively. The level of Bax, Bcl-2 and Cleaved-caspase-3 protein was detected by Western blotting. Results: Compared to the UVB group, the levels of SOD, GSH-Px, Bcl-2 and mitochondrial membrane potentialin the UVB+GSPE group were higher and the levels of MDA, LDH, Bax, Cleaved-caspase-3 apoptosis and ROS were lower. Conclusion: GSPE can protect the HaCaT from oxidative stress induced by UVB.

Key words: HaCaT cells, UVB, grape seed proanthocyanidin extract, oxidative stress, apoptosis