中华皮肤科杂志 ›› 2017, Vol. 50 ›› Issue (7): 512-516.

• 论著 • 上一篇    下一篇

黑果枸杞多糖对中波紫外线诱导HaCaT细胞光损伤的保护作用

任立汆1,加杨娥1,燕华玲1,王永2,郭砚3,哈筱梅1,朱世文1,王刚1   

  1. 1. 青海大学附属医院
    2. 青海省西宁市城西区昆仑路青海大学医学院附属医院皮肤科
    3. 西宁市青海大学附属医院皮肤科
  • 收稿日期:2016-09-12 修回日期:2017-02-28 发布日期:2017-07-03
  • 通讯作者: 燕华玲 E-mail:yhlckw@163.com
  • 基金资助:
    青海柴达木黑果、红果枸杞多糖抑制中波紫外线诱导角质形成细胞(HaCaT)氧化损伤和凋亡及P16基因表达的影响研究

Protective effect of polysaccharides from Lycium ruthenicum Murray against ultraviolet B radiation?induced photodamage in HaCaT cells

  • Received:2016-09-12 Revised:2017-02-28 Published:2017-07-03

摘要: 目的 探讨黑果枸杞多糖对HaCaT细胞光损伤的预防作用及机制。方法 采用超声辅助水提法提取柴达木黑果枸杞中的多糖成分。体外培养的HaCaT细胞分为3组,对照组:正常培养,不做其他处理;中波紫外线(UVB)组:30 mJ/cm2 UVB照射;实验组:30 mJ/cm2 UVB照射前6 h加入2 g/L黑果枸杞多糖溶液。照射1 h后继续培养12 h,倒置显微镜观察细胞形态,MTS法测定细胞增殖,流式细胞仪测定细胞凋亡率,酶标法检测细胞丙二醛和谷胱甘肽过氧化物酶含量、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活力,ELISA法检测上清液和细胞中白细胞介素1(IL?1)和肿瘤坏死因子α(TNF?α)水平。结果 与对照组相比,UVB组细胞形态模糊不清,出现死亡漂浮现象;实验组细胞肿胀,但形态尚清楚。MTS结果显示,3组间细胞增殖吸光度值差异有统计学意义(F = 48.88,P < 0.01),其中UVB组(1.72 ± 0.12)显著低于对照组(2.34 ± 0.11),实验组(2.11 ± 0.10)又显著高于UVB组,差异均有统计学意义(P < 0. 05)。UVB组细胞凋亡率(82.41% ± 2.49%)显著高于对照组(3.98% ± 0.19%)和实验组(22.79% ± 0.97%),实验组细胞凋亡率明显高于对照组,各组间比较差异均有统计学意义(均P < 0.05)。UVB组与对照组比较,丙二醛含量明显上升,SOD活性、谷胱甘肽过氧化物酶含量、CAT活性明显下降(均P < 0.05)。同时,UVB组较对照组上清液中IL?1和TNF?α含量及细胞中TNF?α含量均明显升高(均P < 0.05),但细胞中IL?1差异无统计学意义(P > 0. 05)。结论 黑果枸杞多糖对HaCaT细胞的光损伤有保护作用,其保护机制可能与减少细胞炎症物质的合成和分泌及减少自由基有关。

Abstract: Ren Licuan, Jia Yang′e, Yan Hualing, Wang Yong, Guo Yan, Ha Xiaomei, Zhu Shiwen, Wang Gang Department of Dermatology, Qinghai University Affiliated Hospital, Xining 810000, China Corresponding author: Yan Hualing, Email: yhlckw@163.com 【Abstract】 Objective To evaluate preventive effect of polysaccharides from Lycium ruthenicum Murray against photodamage in HaCaT cells, and to explore its possible mechanism. Methods Ultrasound?assisted extraction was used to extract polysaccharides from Lycium ruthenicum Murray in Qaidam Basin. In vitro cultured HaCaT cells were randomly divided into 3 groups: control group receiving no treatment, ultraviolet B (UVB) group irradiated with 30 mJ/cm2 UVB alone for 1 hour, experimental group pretreated with 2 g/L Lycium ruthenicum polysaccharide solution followed 6 hours later by 30 mJ/cm2 UVB radiation for 1 hour. At twelve hours after UVB radiation, an inverted microscope was used to observe cell morphology. Then, MTS assay was performed to estimate cell proliferation, flow cytometry to detect cell apoptosis, an enzyme?labeled antigen method to detect levels of malondialdehyde (MDA) and glutathione peroxidase (GSH?Px), as well as to evaluate the activity of superoxide dismutase (SOD) and catalase (CAT), and enzyme?linked immunosorbent assay (ELISA) to measure levels of interleukin?1 (IL?1) and tumor necrosis factor?alpha (TNF?α) in HaCaT cells and their culture supernatant. Results Compared with the control group, the UVB group showed obscure cell morphology, cell death and floating phenomenon, while cells became swollen but remained morphologically distinct in the experimental group. MTS assay revealed that the cell proliferative activity significantly differed among the above 3 groups (F = 48.88, P < 0.01), and the cell proliferative activity was significantly lower in the UVB group (1.72 ± 0.12) than in the control group (2.34 ± 0.11, P < 0.05) and experimental group (2.11 ± 0.10, P < 0.05). Moreover, the apoptosis rate was significantly higher in the UVB group (82.41% ± 2.49%) than in the control group (3.98% ± 0.19%, P < 0.05) and experimental group (22.79% ± 0.97%, P < 0.05), as well as higher in the experimental group than in the control group (P < 0.05). Compared with the control group, the UVB group showed significantly higher levels of MDA, supernatant levels of IL?1 and TNF?α, and intracellular levels of TNF?α, but significantly lower GSH?Px levels and activity of SOD and CAT (all P < 0.05). However, there was no signifi?cant difference in the intracellular level of TNF?α between the UVB group and control group (P > 0.05). Conclusion Lycium ruthenicum polysaccharide has protective effects against photodamage in HaCaT cells, likely by reducing the synthesis and secretion of inflammatory substances as well as free radicals.

引用本文

任立汆 加杨娥 燕华玲 王永 郭砚 哈筱梅 朱世文 王刚. 黑果枸杞多糖对中波紫外线诱导HaCaT细胞光损伤的保护作用[J]. 中华皮肤科杂志, 2017,50(7):512-516. doi: