淫羊藿素对人角质形成细胞HaCat辐射损伤的保护作用

辐射防护 ›› 2019, Vol. 39 ›› Issue (4): 338-344.

淫羊藿素对人角质形成细胞HaCat辐射损伤的保护作用

王锃, 洪金省, 陈金梅, 吴建东, 苏丽

福建医科大学附属第一医院,放射生物福建省高等学校重点实验室,福州 350005

收稿日期:2018-12-04 出版日期:2019-07-15 发布日期:2019-10-29
通讯作者: 苏丽。E-mail:lily2230@126.com
作者简介:王锃(1984—),男,2007年毕业于华侨大学生物工程专业,2013年博士毕业于厦门大学动物学专业,研究实习员。E-mail:wzishere@126.com
基金资助:
福建省自然科学基金面上项目(2016J01541,2017J01184)。

Protection effects of icaritin on HaCat cells of human skin radiation damage

WANG Zeng, HONG Jinsheng, CHEN Jinmei, WU Jiandong, SU Li

The First Affiliated Hospital of Fujian Medical University, Key Lab of Radiation Biology ofFujian Province Universities, Fuzhou 350005

Received:2018-12-04 Online:2019-07-15 Published:2019-10-29

摘要/Abstract

摘要： 研究淫羊藿素对人角质形成细胞HaCat的放射防护作用。将HaCat细胞分成正常对照组、单纯淫羊藿素组、单纯照射组以及照射+淫羊藿素高/中/低剂量组。以6 MV X射线(剂量率为0.5 Gy/min),给予照射组和各个加药照射组细胞20 Gy的单次照射。所有药物处理组均于照射前6小时给予不同浓度的淫羊藿素处理,细胞照射后1小时,采用DCFH-DA探针检测细胞内活性氧水平、6小时使用实时荧光定量PCR法测定炎症因子mRNA表达水平、24小时采用硫代巴比妥酸法检测胞内丙二醛水平、48小时CCK-8法检测细胞的增殖能力、Annexin V-PI双染检测细胞凋亡、酶联免疫吸附法测定炎症因子分泌水平。结果表明,HaCat细胞经20 Gy的X射线照射后,细胞生存率下降,细胞凋亡率升高。给予淫羊藿素处理,可以刺激其增殖,提高细胞生存率,降低细胞凋亡比例。照射后细胞内活性氧(ROS)和丙二醛(MDA)的含量显著升高,淫羊藿素处理可以降低两者的含量,保护细胞。同时受到照射的HaCat细胞炎症因子IL-1β、IL-6以及TNF-α表达和分泌的量显著升高,淫羊藿素可以抑制这些因子的表达,减少其分泌。实验结果证明,淫羊藿素对HaCat细胞存在明显的放射保护作用,具有促进增殖、抑制凋亡、抗氧化和抗炎的能力,有望成为防护放射性皮肤损伤的中药防护剂。

关键词: 淫羊藿素, HaCat细胞, 放射性皮肤损伤

Abstract: The study of the radiological protective effect of icaritin on HaCat cells was carried out. CCK-8 method was used to detect the change of cell proliferation; Annexin V-PI double staining was used to detect cell apoptosis; DCFH-DA probe was used to detect reactive oxygen species and thiobarbituric acid method was used to measure intracellular malondialdehyde levels. Inflammatory factor secretion levels were determined by real-time PCR and Elisa assay. After the HaCat cells were treated with 20 Gy of X-ray, the cell viability was decreased and apoptosis rate was increased. It was found that treatment with icaritin can stimulate its proliferation by increasing cell survival rate and reducing the proportion of apoptosis. The content of ROS and MDA in the cells increased significantly after irradiation, but the treatment with icaritin could reduce the content of both and protect the cells. Meanwhile, the amount of inflammatory factors IL-1β, IL-6 and TNF-α expressed and secreted by HaCat cells was significantly increased, and icaritin inhibited the expression of these factors and reduced the amount of secretion. The results indicated that icaritin had obvious radiological protective effect on HaCat cells. It has the ability to promote proliferation, inhibit apoptosis, anti-oxidation and anti-inflammatory. It is expected to be a protective agent for traditional Chinese medicine against radiation-induced skin damage.

Key words: icaritin, HaCat cells, radiation-induced skin damage

中图分类号:

R142+.4

王锃, 洪金省, 陈金梅, 吴建东, 苏丽. 淫羊藿素对人角质形成细胞HaCat辐射损伤的保护作用[J]. 辐射防护, 2019, 39(4): 338-344.

WANG Zeng, HONG Jinsheng, CHEN Jinmei, WU Jiandong, SU Li. Protection effects of icaritin on HaCat cells of human skin radiation damage[J]. RADIATION PROTECTION, 2019, 39(4): 338-344.

参考文献

[1] Chan R J, Webster J, Chung B, et al. Prevention and treatment of acute radiation-induced skin reactions: a systematic review and meta-analysis of randomized controlled trials[J]. BMC Cancer, 2014, 14(1):53. DOI: 10.1186/1471-2407-14-53.
[2] 刘松, 刘超明, 赖丽娟, 等. 淫羊藿素的药理作用研究进展[J]. 赣南医学院学报, 2017, 37(4): 631-635. DOI: 10.3969/j.issn.1001-5779.2017.04.044.
LIU Song, LIU Chaoming, LAI Lijuan, et al. Research progress in pharmacological effects of icaritin[J]. Journal of Gannan Medical University, 2017, 37(4): 631-635. DOI: 10.3969/j.issn.1001-5779.2017.04.044.
[3] 韩冰,杨峻山.淫羊藿药理作用研究概况[J].中草药, 2000, (11):873-875. DOI: 10.3321/j.issn:0253-2670.2000.11.039.
HAN Bing, YANG Junshan. Advances in pharmacological studies on the effects of epimedium[J]. Chinese Traditional and Herbal Drugs, 2000, (11):873-875. DOI: 10.3321/j.issn:0253-2670.2000.11.039.
[4] GAO Lvfen,CHEN Ming,OUYANG Yuan, et al. Icaritin induces ovarian cancer cell apoptosis through activation of p53 and inhibition of Akt/mTOR pathway[J]. Life Sciences, 2018:S0024320518301814. DOI: 10.1016/j.lfs.2018.03.059.
[5] LI Hongchao, LIANG Qinghua, WANG Lin. Icaritin inhibits glioblastoma cell viability and glycolysis by blocking the IL-6/Stat3 pathway[J].Journal of Cellular Biochemistry, 2019, 120 (5): 7 257-7 264. DOI: 10.1002/jcb.28000.
[6] YANG, Xiaojing, XI Yaming, LI Zijian. Icaritin: A novel natural candidate for hematological malignancies therapy[J]. Bio Med Research International, 2019: 4860268. DOI: 10.1155/2019/4860268.
[7] 王红丽,刘效栓,沈涛,等. 脱水淫羊藿素对老年大鼠的抗氧化作用[J].中成药, 2017, 39(8):1698-1700. DOI:10.3969/j.issn.1001-1528.2017.08.033.
WANG Hongli, LIU Xiaoshuan, SHEN Tao,et al. Antioxidant effect of dehydrated icariin on aged rats [J]. Chinese Traditional Patent Medicine, 2017, 39(8):1 698-1 700. DOI:10.3969/j.issn.1001-1528.2017.08.033.
[8] 熊吻, 甘振宝, 范江华, 等. 淫羊藿素对大鼠骨髓间充质干细胞增殖的影响[J]. 广西中医药, 2018, 41(235):54-56. DOI: 10.3969/j.issn.1003-0719.2018.01.023.
XIONG Wen, GAN Zhenbao, FAN Jianghua, et al. Effect of Icariin on proliferation of rat bone marrow mesenchymal stem cells [J]. Guangxi Journal of Traditional Chinese Medicine, 2018, 41(235):54-56. DOI: 10.3969/j.issn.1003-0719.2018.01.023.
[9] 马山, 麻培培, 谌文元, 等. 基于抗氧化作用的淫羊藿抗衰老药效研究(英文)[J]. 中国现代应用药学, 2017, 34(6): 836-840. DOI: 10.13748/j.cnki.issn1007-7693.2017.06.011.
MA Shan,MA Peipei,CHEN Wenyuan, et al. Anti-aging effects of Epimedium Acuminatum based on the anti-oxidant effect [J]. Chin J Mod Appl Pharm,2017, 34(6):836-840. DOI: 10.13748/j.cnki.issn1007-7693.2017.06.011.
[10] 姜明春. 淫羊藿苷及淫羊藿素通过GPER和IGF-1R抑制小胶质细胞炎症反应保护多巴胺能神经元的实验研究[D]. 青岛大学, 2017.
[11] ZHANG Wei, XING Baichun, YANG Linlin, et al. Icaritin attenuates myocardial ischemia and reperfusion injury via anti-inflammatory and anti-oxidative stress effects in rats[J]. The American Journal of Chinese Medicine, 2015:1-15. DOI: 10.1142/S0192415X15500627.
[12] SHAN Shibing, YAO Yanliang, XU Jingchun, et al. Preparation of icaritin-loaded mPEG-PLA micelles and evaluation on ischemic brain injury[J]. Journal of Biomedical Nanotechnology, 2019, 15(4): 674-685. DOI: 10.1166/jbn.2019.2721.
[13] Yarnold J, Brotons M C. Pathogenetic mechanisms in radiation fibrosis[J]. Radiotherapy & Oncology, 2010, 97(1): 149-161. DOI: 10.1016/j.radonc.2010.09.002.
[14] Terrazzino S, Deantonio L, Cargnin S, et al. DNA methyltransferase gene polymorphisms for prediction of radiation-induced skin fibrosis after treatment of breast cancer: a multifactorial genetic approach[J]. Cancer Research & Treatment, 2017, 49(2): 464-472. DOI: 10.4143/crt.2016.256.