枸杞酰胺体外对镉致成骨细胞凋亡的保护作用

doi:10.11843/j.issn.0366-6964.2023.03.038

畜牧兽医学报 ›› 2023, Vol. 54 ›› Issue (3): 1273-1280.doi: 10.11843/j.issn.0366-6964.2023.03.038

枸杞酰胺体外对镉致成骨细胞凋亡的保护作用

常颖艳^1,3, 赵鸿雁^2,3, 胡茂志^1,3, 刘宗平^2,3*

1. 扬州大学生物科学与技术学院, 扬州 225009;
2. 扬州大学兽医学院, 扬州 225009;
3. 江苏高校动物重要疫病与人兽共患病防控协同创新中心, 扬州 225009

收稿日期:2022-08-08 出版日期:2023-03-23 发布日期:2023-03-21
通讯作者: 刘宗平,主要从事动物营养代谢病与中毒病研究,E-mail:liuzongping@yzu.edu.cn
作者简介:常颖艳(1997-)，女，江苏丹阳人，硕士生，主要从事动物营养代谢病与中毒病研究，E-mail：1932469462@qq.com
基金资助:
国家自然科学基金资助项目(32172923)；江苏省高等学校自然科学研究重大项目(20KJA230002)；江苏高校优势学科建设工程资助项目(PAPD)

Protective Effect of Aurantiamide Acetate on Cd-induced Apoptosis of Osteoblasts in vitro

CHANG Yingyan^1,3, ZHAO Hongyan^2,3, HU Maozhi^1,3, LIU Zongping^2,3*

1. College of Bioscience and Biotechnology, Yangzhou University, Yangzhou 225009, China;
2. College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China;
3. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China

Received:2022-08-08 Online:2023-03-23 Published:2023-03-21

摘要/Abstract

摘要： 旨在探究枸杞酰胺(aurantiamide acetate, AA)在镉(Cd)致成骨细胞凋亡中的保护效应，本研究以ROS1728大鼠成骨细胞系为研究对象，添加5 μg·mL^-1 AA预处理3 h后，加入20 μg·mL^-1氯化镉(CdCl₂)继续处理6 h。利用流式细胞术检测细胞凋亡率、线粒体膜电位(mitochondrial membrane potential, MMP)、活性氧簇(reactive oxygen species, ROS)，Western blot 检测Bcl-2、Bax蛋白，高内涵细胞成像分析系统观察和分析细胞线粒体纹理结构和指数变化。结果显示，经过AA预处理后，较Cd处理组， AA极显著抑制Cd导致的ROS1728细胞凋亡率上升，Bax/Bcl-2比值增高，MMP下降，线粒体纹理结构指数下降，ROS含量增多(P<0.01)。上述结果表明，AA通过降低细胞内过氧化物含量，维持细胞线粒体结构的完整，抑制镉诱导的ROS1728细胞凋亡。

关键词: 镉, 枸杞酰胺, ROS1728, 凋亡, 线粒体, ROS

Abstract: This study aimed to explore the protective effect of aurantiamide acetate (AA) on cadmium (Cd)-induced apoptosis of osteoblasts. ROS1728 cells were used as osteoblast model. ROS1728 cells were pretreated with 5 μg·mL^-1 AA for 3 h, and then treated with 20 μg·mL^-1 CdCl₂ for 6 h. Flow cytometry was applied to detect cell apoptotic rate, mitochondrial membrane potential (MMP), and content of reactive oxygen species (ROS). Western blot was used to detect the expression of Bcl-2 and Bax. After Mito-tracker Red staining, High Content Analysis System (HCAS) was used to observe and analyze cellular mitochondrial texture index and structural changes. The results showed that compared with Cd treatment group, AA pretreatment significantly inhibited Cd-induced apoptosis rate increase Bax / Bcl-2 ratio rise, MMP decrease, mitochondrial texture structure index decline and ROS accumulation (P<0.01). The results showed that AA could inhibit Cd-induced apoptosis of ROS1728 cells by maintaining the integrity of mitochondrial structure via reducing the content of peroxide.

Key words: cadmium, aurantiamide acetate, ROS1728, apoptosis, mitochondrion, ROS

中图分类号:

常颖艳, 赵鸿雁, 胡茂志, 刘宗平. 枸杞酰胺体外对镉致成骨细胞凋亡的保护作用[J]. 畜牧兽医学报, 2023, 54(3): 1273-1280.

CHANG Yingyan, ZHAO Hongyan, HU Maozhi, LIU Zongping. Protective Effect of Aurantiamide Acetate on Cd-induced Apoptosis of Osteoblasts in vitro[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 1273-1280.

参考文献 28

[1]	黄吉厚.镉对环境的污染及其对策研究[J].辽宁教育学院学报, 2000, 17(5):29-31.HUANG J H. Studies on environmental pollution of cadmium and its countermeasures[J]. Journal of Shenyang Normal University Educational Science Edition, 2000, 17(5):29-31.(in Chinese)
[2]	NORDBERG G. Cadmium and human health:a perspective based on recent studies in China[J]. J Trace Elem Exp Med, 2003, 16(4):307-319.
[3]	ENGSTRÖM A, SKERVING S, LIDFELDT J, et al. Cadmium-induced bone effect is not mediated via low serum 1, 25-dihydroxy vitamin D[J]. Environ Res, 2009, 109(2):188-192.
[4]	CHEN X, REN S, ZHU G Y, et al. Emodin suppresses cadmium-induced osteoporosis by inhibiting osteoclast formation[J]. Environ Toxicol Pharmacol, 2017, 54:162-168.
[5]	XIE D D, SHENG Z F. Low-level cadmium exposure and bone health[J]. J Bone Miner Res, 2017, 32(2):419.
[6]	MA Y G, RAN D, ZHAO H Y, et al. Cadmium exposure triggers osteoporosis in duck via P2X7/PI3K/AKT-mediated osteoblast and osteoclast differentiation[J]. Sci Total Environ, 2021, 750:141638.
[7]	ZHAO H Y, LIU W, WANG Y, et al. Cadmium induces apoptosis in primary rat osteoblasts through caspase and mitogen-activated protein kinase pathways[J]. J Vet Sci, 2015, 16(3):297-306.
[8]	ZHENG J M, ZHUO L L, RAN D, et al. Cadmium induces apoptosis via generating reactive oxygen species to activate mitochondrial p53 pathway in primary rat osteoblasts[J]. Toxicology, 2020, 446:152611.
[9]	马天宇,王磊,金铃和,等.地骨皮化学成分及生物活性研究[J].中国果菜, 2018, 38(5):9-13.MA T Y, WANG L, JIN L H, et al. Research progress on chemical constituents and biological actions of cortex Lycii[J]. China Fruit&Vegetable, 2018, 38(5):9-13.(in Chinese)
[10]	YOON C S, KIM D C, LEE D S, et al. Anti-neuroinflammatory effect of aurantiamide acetate from the marine fungus Aspergillus sp. SF-5921:inhibition of NF-κB and MAPK pathways in lipopolysaccharide-induced mouse BV2 microglial cells[J]. Int Immunopharmacol, 2014, 23(2):568-574.
[11]	LIU X B, YANG B X, ZHANG L, et al. An in vivo and in vitro assessment of the anti-inflammatory, antinociceptive, and immunomodulatory activities of Clematis terniflora DC. extract, participation of aurantiamide acetate[J]. J Ethnopharmacol, 2015, 169:287-294.
[12]	BISSIM S M, KENMOGNE S B, LOBE J S, et al. The chemistry and biological activities of Citrus clementina Hort. Ex Tanaka (Rutaceae), a vegetatively propagated species[J]. Nat Prod Res, 2021, 35(22):4839-4842.
[13]	胡永美,叶文才,殷志琦,等.芝麻花化学成分的研究[J].药学学报, 2007, 42(3):286-291.HU Y M, YE W C, YIN Z Q, et al. Chemical constituents from Flos Sesamum indicum L.[J]. Acta Pharmaceutica Sinica, 2007, 42(3):286-291.(in Chinese)
[14]	DENG Y, DING T T, DENG L L, et al. Active constituents of Zanthoxylum nitidium from Yunnan Province against leukaemia cells in vitro[J]. BMC Chem, 2021, 15(1):44.
[15]	SASHIDHARA K V, ROSAIAH J N, TYAGI E, et al. Rare dipeptide and urea derivatives from roots of Moringa oleifera as potential anti-inflammatory and antinociceptive agents[J]. Eur J Med Chem, 2009, 44(1):432-436.
[16]	赵伟杰.乌苏里藜芦中橙黄胡椒酰胺的分离鉴定[J].中国中药杂志, 1998, 23(1):41, 63.ZHAO W J. Isolation and structure determination of aurantiamide acetate from Veratrum nigrum L. var. ussuriense Nakai[J]. China Journal of Chinese Materia Medica, 1998, 23(1):41, 63.(in Chinese)
[17]	ALVES C Q, LIMA L S, DAVID J M, et al. In vitro acetylcholinesterase activity of peptide derivatives isolated from two species of Leguminosae[J]. Pharm Biol, 2013, 51(7):936-939.
[18]	YANG Y, ZHANG L H, YANG B X, et al. Aurantiamide acetate suppresses the growth of malignant gliomas in vitro and in vivo by inhibiting autophagic flux[J]. J Cell Mol Med, 2015, 19(5):1055-1064.
[19]	WANG H W, GAO S F, LI J M, et al. Hybrids of aurantiamide acetate and isopropylated genipin as potential anti-inflammatory agents:The design, synthesis, and biological evaluation[J]. Chem Biol Drug Des, 2021, 97(4):797-808.
[20]	余超,张维蔚,梁伯衡,等.广州市市售普洱茶镉元素残留现状及分布差异[J].职业与健康, 2016, 32(9):1208-1211.YU C, ZHANG W W, LIANG B H, et al. Cadmium residual status in commercially available Pu'er tea and its distribution difference in Guangzhou City[J]. Occupation and Health, 2016, 32(9):1208-1211.(in Chinese)
[21]	MANOLAGAS S C. Birth and death of bone cells:Basic regulatory mechanisms and implications for the pathogenesis and treatment of osteoporosis[J]. Endocr Rev, 2000, 21(2):115-137.
[22]	LV Y J, WANG P, HUANG R, et al. Cadmium exposure and osteoporosis:a population-based study and benchmark dose estimation in Southern China[J]. J Bone Miner Res, 2017, 32(10):1990-2000.
[23]	MA Y G, RAN D, SHI X N, et al. Cadmium toxicity:a role in bone cell function and teeth development[J]. Sci Total Environ, 2021, 769:144646.
[24]	LIZOTTE J, ABED E, SIGNOR C, et al. Expression of macrophage migration inhibitory factor by osteoblastic cells:protection against cadmium toxicity[J]. Toxicol Lett, 2012, 215(3):167-173.
[25]	冯居平,符保军.中药内外合治骨痨26例[J].河南中医学院学报, 2007, 22(4):71-72.FENG J P, FU B J. 26 cases of bone TB with therapeutical order of Chinese medicine[J]. Journal of Henan University of Chinese Medicine, 2007, 22(4):71-72.(in Chinese)
[26]	李玉萍.黄芪寄生汤治疗肝肾气血亏虚型膝骨关节炎的临床体会[J].人人健康, 2022(13):87-89.LI Y P. Clinical experience of Huang Qi Jisheng decoction in the treatment on knee osteoarthritis with Liver-Kidney Qi and blood deficiency type of syndrome[J]. Health for Everyone, 2022(13):87-89.(in Chinese)
[27]	OZBEN B, ERDOGAN O. The role of inflammation and allergy in acute coronary syndromes[J]. Inflamm Allergy Drug Targets, 2008, 7(3):136-144.
[28]	ZHOU B X, YANG Z F, FENG Q T, et al. Aurantiamide acetate from baphicacanthus cusia root exhibits anti-inflammatory and anti-viral effects via inhibition of the NF-κB signaling pathway in Influenza A virus-infected cells[J]. J Ethnopharmacol, 2017, 199:60-67.

枸杞酰胺体外对镉致成骨细胞凋亡的保护作用

Protective Effect of Aurantiamide Acetate on Cd-induced Apoptosis of Osteoblasts in vitro

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