Puerarin protects rat cervical intervertebral disc annulus fibrosus cells: An optimal concentration study

doi:10.3969/j.issn.2095-4344.2013.07.004

Abstract

Abstract:

BACKGROUND: Puerarin restrains the apoptosis of disc annulus fibrosus cells and delays the degenerationof the intervertebral disc.
OBJECTIVE: To explore the protective role of puerarin in rat cervical intervertebral disc annulus fibrosus cells.
METHODS: Cervical intervertebral disc annulus fibrosus cells were induced by different concentrations of Fas ligand (5, 20, 50 μg/L). The appropriate concentration of puerarin was detected using Annexin V-FITC-PI double staining to establish an in vitro apoptotic model of annulus fibrosus cells. Changes of annulus fibrosus cells in response to different concentrations of puerarin (0.01, 0.1, 1 g/L) were observed by flow cytometry and MTT method.
RESULTS AND CONCLUSION: 5 μg/L Fas ligand had no significant influence on the survival rate and early-apoptotic rate of annulus fibrosus cells (P > 0.05). 20 and 50 μg/L Fas ligand significantly promoted annulus fibrosus cells apoptosis (P < 0.01) in a dose-dependent manner. The 0.01, 0.1 and 1 g/L puerarin could improve the survival rate of Fas ligand-induced annulus fibrosus cells in a dose-dependent manner (P < 0.01). These findings indicate that (1) Fas ligand can induce the disc annulus fibrosus to be in apoptosis status. (2) Puerarin (0.01, 0.1, 1 g/L) can promote the proliferation of disc annulus fibrosus cells in a dose-dependent manner. (3) The apoptosis rate of disc annulus fibrosus cells induced by Fas ligand in vitro can be restrained by puerarin interference.

Key words: tissue construction, cartilage tissue construction, cervical spondylosis, puerarin, Chinese herbs, optimal concentration, dose-dependent, Fas/Fas ligand, cell necrosis, annulus fibrosus cells, rats, the National Natural Science Foundation of China, tissue construction photographs-containing paper

CLC Number:

R318

Chen Shao-qing, Lin Jian-ping, Wang Shi-zhong, Chen Le-chun, Hong Yu, Zhang Kun-mu. Puerarin protects rat cervical intervertebral disc annulus fibrosus cells: An optimal concentration study[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(7): 1156-1161.

Figures/Tables 8

References

[1] Wang SZ,Chen SJ,Song HM. Zhongguo Kangfu Lilun yu Shijian. 2005;11(11):956. 王诗忠,陈水金,宋红梅.颈椎间歇性拔伸手法治疗青年颈椎病的疗效分析[J].中国康复理论与实践,2005,11(11):956.

[2] Adams MA, Roughley PJ.What is intervertebral disc degeneration, and what causes it. Spine (Phila Pa 1976). 2006;31(18):2151-2161.

[3] Zhao CQ, Jiang LS, Dai LY.Programmed cell death in intervertebral disc degeneration.Apoptosis. 2006;11(12): 2079-2088.

[4] Park JB, Kim KW, Han CW,et al. Expression of Fas receptor on disc cells in herniated lumbar disc tissue.Spine (Phila Pa 1976). 2001;26(2):142-146.

[5] Park JB, Chang H, Kim KW.Expression of Fas ligand and apoptosis of disc cells in herniated lumbar disc tissue.Spine (Phila Pa 1976). 2001;26(6):618-621.

[6] Rannou F, Lee TS, Zhou RH,et al. Intervertebral disc degeneration: the role of the mitochondrial pathway in annulus fibrosus cell apoptosis induced by overload.Am J Pathol. 2004;164(3):915-924.

[7] Cui LY, Liu SL, Ding Y,et al. IL-1beta sensitizes rat intervertebral disc cells to Fas ligand mediated apoptosis in vitro.Acta Pharmacol Sin. 2007;28(10):1671-1676.

[8] Hadjipavlou AG, Tzermiadianos MN, Bogduk N,et al. The pathophysiology of disc degeneration: a critical review.J Bone Joint Surg Br. 2008;90(10):1261-1270.

[9] Wang YJ,Shi Q,Li JS,et al. Zhongguo Kangfu Yixue Zazhi. 2003;18(7):403-406. 王拥军,施杞,李家顺,等.椎间盘细胞信号传导及益气化瘀方调控机理的研究[J].中国康复医学杂志,2003,18(7):403-406.

[10] Yang J. Liaoning Zhongyi Zazhi. 2004;31(7):559. 杨军.颈椎病常用内服中药选配规律[J].辽宁中医杂志,2004, 31(7): 559.

[11] Zhao GC,Li CR. Sichuan Shengli Kexue Zazhi. 2009;31(1): 24-25. 赵桂昌,李才锐.葛根素注射液对颈椎病的临床观察[J].四川生理科学杂志, 2009,31(1):24-25.

[12] Song YJ. Yunnan Zhongyi Zhongyao Zazhi. 2001;22(3): 20-21. 宋云娟.葛根素治疗颈椎病的临床观察[J].云南中医中药杂志, 2001,22(3):20-21.

[13] Qu SH. Xinjiang Zhongyiyao. 2005;23(5):28-29. 曲少华.葛根素配合针灸电脑中频治疗颈椎病31例[J].新疆中医药,2005,23(5):28-29.

[14] Yang SH. Hebei Zhongyi. 2006;28(10):746. 杨绍华.葛根素合中药熏蒸治疗颈椎病临床观察[J].河北中医, 2006,28(10):746.

[15] Jiang DH,Zhang AH. Zhongguo Zhongyi Jichu Yixue Zazhi. 2004;10(10):759-761. 姜东辉,张爱华.葛根素配合手法治疗颈椎病62例临床疗效观察[J].中国中医基础医学杂志,2004,10(10):759-761.

[16] Zou QB. Zhongguo Xiandai Yaowu Yingyong. 2010;4(12): 382-384. 邹清波.葛根素对心血管系统的药用价值[J].中国现代药物应用, 2010,4(12):382-384.

[17] Zhou Q,Wang YJ,Shi Q. Jizhu Waike Zazhi. 2003;1(4): 226-229. 周泉,王拥军,施杞.椎间盘纤维环细胞的培养[J].脊柱外科杂志, 2003,1(4):226-229.

[18] Cui LY, Liu SL, Ding Y,et al. IL-1beta sensitizes rat intervertebral disc cells to Fas ligand mediated apoptosis in vitro. Acta Pharmacol Sin. 2007;28(10):1671-1676.

[19] Wang WT,Hao ML,Wang FY,et al. Zhongguo Bingli Shengli Zazhi. 2006;22(8):1575-1580. 王万铁,郝卯林,王方岩,等.葛根素对肺缺血-再灌注损伤时Fas/FasL表达的影响[J].中国病理生理杂志,2006,22(8): 1575-1580.

[20] Wen J,Chen SL,Xu HX. Xiandai Zhongyao Yanjiu yu Shijian. 2006;20(1):61-64. 温葭,陈士林,徐宏喜.葛根素对细胞凋亡影响的研究[J].现代中药研究与实践,2006,20(1):61-64.

[21] Han JQ,Li GC,Zhou XL,et al. Zhongxiyi Jiehe Xinnao Xueguanbing Zazhi. 2008;6(10):1179-1181. 韩江全,李官成,周晓兰,等.葛根素对大鼠局灶性脑缺血再灌注后神经细胞凋亡及Bcl-2、Bax蛋白表达的影响[J].中西医结合心脑血管病杂志,2008,6(10):1179-1181.

[22] Xiao X,Mei GL,Hu WD,et al. Jiaotong Yixue. 2008;22(5): 472-473. 肖旭,梅广林,胡卫东,等.葛根素对大鼠肝缺血再灌注后Caspase-3和Bcl-2表达的影响[J].交通医学,2008,22(5): 472-473.