抗坏血酸-聚乙烯亚胺复合碳点通过高尔基体应激对MG63细胞增殖、凋亡和氧化应激的影响

doi:10.13481/j.1671-587x.20190605

吉林大学学报(医学版) ›› 2019, Vol. 45 ›› Issue (06): 1218-1223.doi: 10.13481/j.1671-587x.20190605

抗坏血酸-聚乙烯亚胺复合碳点通过高尔基体应激对MG63细胞增殖、凋亡和氧化应激的影响

孟许亚¹, 刘杰^2,3, 王璐^2,3, 布文奂^2,3, 卢金金⁴, 孙宏晨⁵

1. 陕西省西安市第五医院口腔科, 陕西西安 710000;
2. 吉林省牙发育及颌骨重塑与再生重点实验室, 吉林长春 130021;
3. 吉林大学口腔医院口腔病理科, 吉林长春 130021;
4. 吉林大学口腔医院儿童口腔科, 吉林长春 130021;
5. 中国医科大学附属口腔医院口腔病理科, 辽宁沈阳 110000

收稿日期:2019-02-17 出版日期:2019-12-05 发布日期:2019-12-05
通讯作者: 孙宏晨,教授,主任医师,博士研究生导师(Tel:0431-88796010,E-mail:hcsun@mail.jlu.edu.cn) E-mail:hcsun@mail.jlu.edu.cn
作者简介:孟许亚(1991-),女,河南省汝州市人,医师,医学硕士,主要从事纳米材料促进成骨细胞分化方面的研究。
基金资助:
国家自然科学基金资助课题（81320108011）；吉林省科技厅自然科学基金资助课题（20170101093JC）；吉林省省级产业创新专项资金资助课题（2016C044-3）

Effects of ascorbic acid-polyethyleneimine carbon dots on proliferation, apoptosis and oxidative stress of MG63 cells by Golgi stress

MENG Xuya¹, LIU Jie^2,3, WANG Lu^2,3, BU Wenhuan^2,3, LU Jinjin⁴, SUN Hongchen⁵

1. Department of Dentistry, Fifth Hospital, Xi'an City, Shaanxi Province, Xi'an 710000, China;
2. Key Laboratory of Tooth Development and Bone Remodeling and Regeneration of Jilin Province, Changchun 130021, China;
3. Department of Oral Pathology, Stomatology Hospital, Jilin University, Changchun 130021, China;
4. Department of Pediatric Dentistry, Stomatology Hospital, Jilin University, Changchun 130021, China;
5. Department of Oral Pathology, Affiliated Stomatology Hospital, China Medical University, Shenyang 110000, China

Received:2019-02-17 Online:2019-12-05 Published:2019-12-05

摘要/Abstract

摘要： 目的：探讨抗坏血酸-聚乙烯亚胺复合碳点（CDs）对人骨肉瘤MG63细胞增殖、凋亡和氧化应激等的影响，阐明抗坏血酸-聚乙烯亚胺复合CDs的细胞生物学特性。方法：以抗坏血酸和聚乙烯亚胺为原料，用微波法合成抗坏血酸-聚乙烯亚胺复合CDs。采用MTT法检测MG63细胞增殖率，流式细胞术检测不同细胞周期MG63细胞百分比。将MG63细胞分为空白对照组、转染阴性对照组、转染GOLPH3-siRNA组、CDs组、转染阴性对照+CDs组、转染GOLPH3-siRNA+CDs组。采用RT-PCR法检测各组MG63细胞中高尔基体磷酸化蛋白3（GOLPH3）的基因表达水平并计算基因沉默效率，流式细胞术检测各组MG63细胞凋亡率以及MG63细胞中活性氧（ROS）水平。结果：与空白对照组比较，随着CDs浓度的增加，MG63细胞的增殖率降低；在24和48 h时，CDs浓度大于40 mg·L^-1时，MG63细胞增殖率明显降低（P<0.05）；在72 h，CDs浓度大于20 mg·L^-1时，细胞增殖率明显降低（P<0.05）。与对照组比较，40 mg·L^-1CDs组G₀/G₁期MG63细胞百分比明显升高（P<0.01），S期MG63细胞百分比明显降低（P<0.01）。倒置荧光显微镜下CDs具有一定的光致发光特性；与非碳点组（空白对照组、转染阴性对照组、转染GOLPH3-siRNA组）比较，碳点组（CDs组、转染阴性对照+CDs组、转染GOLPH3-siRNA+CDs组） MG63细胞凋亡率和细胞中ROS水平升高（P<0.05）。结论：CDs具有低毒性和光致发光特性，可以阻滞细胞周期，并促进细胞凋亡和ROS生成。GOLPH3具有抗凋亡和抑制ROS生成的作用，对细胞的存活起到一定的保护作用。

关键词: 碳点, 抗坏血酸, 聚乙烯亚胺, 高尔基体应激, 细胞增殖, 细胞凋亡

Abstract: Objective: To investigate the effects of ascorbic acid-polyethyleneimine carbon dots(CDs) on the proliferation, apoptosis and oxidative stress of the human osteosarcoma cell line MG63 cells, and to elucidate the biological characteristics of ascorbic acid-polyethyleneimine CDs. Methods: The ascorbic acid-polyethyleneimine CDs were synthesized with ascorbic acid and polyethyleneimine by microwave method. The proliferation rate of MG63 cells was detected by MTT method, and the percentages of MG63 cells at different cell cycles were detected by flow cytometry.The MG63 cells were divided into blank control group,negative control transfection group, GOLPH3-siRNA group, CDs group,negative control transfection+CDs group, GOLPH3-siRNA+CDs group. The expression levels of Golgi phosphorylation protein 3 (GOLPH3) gene in the MG63 cells in various groups were detected by PT-PCR method and the gene silencing efficiency was calculated; the apoptotic rates and reactive oxygen specie(ROS)levels in the MG63 cells in various groups were detected by flow cytometry. Results: Compared with blank control group, the proliferation rates of the MG63 cells were decreased with the increasing of CDs concentration; at 24 and 48 h, when the CDs concentration was more than 40 mg·L^-1, the proliferation rates of the MG63 cells were decreased significantly (P<0.05); at 72 h, when the CDs concentration was more than 20 mg·L^-1, the proliferation rates of the MG63 cells were decreased significantly (P<0.05). Compared with control group, the percentage of the MG63 cells at G₀/G₁ phase in CDs group was increased significantly(P<0.01), the percentage of the MG63 cells at S phase in CDs group was decreased significantly(P<0.01). The photoluminescence properties of CDs in the MG63 cells were seen under inverted fluorescence microscope; compared with non-CDs groups (blank control group, negative control transfection group, GOLPH3-siRNA group),the apoptotic rates of MG63 cells and the ROS levels in the MG63 cells in CDs groups(CDs group,negative control transfection+CDs group, GOLPH3-siRNA+CDs group)were increased(P<0.05). Conclusion: CDs have low toxicity and photoluminescence properties,which can arrest the cell cycle and promote the apoptosis and ROS production.GOLPH3 has the anti-apoptosis effect and inhibitory effect on the production of ROS to protect the survival of cells.

Key words: carbon dots, ascorbic acid, polyethyleneimine, Golgi stress, cell proliferation, apoptosis

中图分类号:

R329.2

孟许亚, 刘杰, 王璐, 布文奂, 卢金金, 孙宏晨. 抗坏血酸-聚乙烯亚胺复合碳点通过高尔基体应激对MG63细胞增殖、凋亡和氧化应激的影响[J]. 吉林大学学报(医学版), 2019, 45(06): 1218-1223.

MENG Xuya, LIU Jie, WANG Lu, BU Wenhuan, LU Jinjin, SUN Hongchen. Effects of ascorbic acid-polyethyleneimine carbon dots on proliferation, apoptosis and oxidative stress of MG63 cells by Golgi stress[J]. Journal of Jilin University(Medicine Edition), 2019, 45(06): 1218-1223.

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抗坏血酸-聚乙烯亚胺复合碳点通过高尔基体应激对MG63细胞增殖、凋亡和氧化应激的影响

Effects of ascorbic acid-polyethyleneimine carbon dots on proliferation, apoptosis and oxidative stress of MG63 cells by Golgi stress

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