益气化瘀补肾方对退变椎间盘细胞蛋白聚糖和Ⅹ型胶原mRNA表达的影响

doi:10.3736/jcim20111016

Abstract

Objective: To investigate the effects of serum containing Yiqi Huayu Bushen Recipe, a compound Chinese herbal medicine, on expressions of aggrecan and type Ⅹ collagen mRNAs of the degenerated intervertebral disc cells in patients with cervical spondylotic myelopathy.

Methods: Six inpatients diagnosed as cervical spondylotic myelopathy and undergoing cervical decompression and fusion surgery to remove the degenerated disc tissue were enrolled in this study. Primary intervertebral disc cells were isolated using small tissue mass method. Serum containing Yiqi Huayu Bushen recipe were obtained after SD rats were fed with Yiqi Huayu Bushen recipe for 3 days. MTT method was used to detect proliferation of the first-passage cells isolated from degenerated intervertebral discs and growth curve was drawn. In the following tests, the first-passage cells were cultured with the sera containing drugs and sodium chloride (NaCl), respectively. Proliferation of intervertebral disc cells was detected by MTT method and expressions of aggrecan and type Ⅹ collagen mRNAs were detected by real-time polymerase chain reaction.

Results: The growth curve of the first-passage cells of the degenerated intervertebral discs showed that the time of cell proliferation peak was the sixth day. Compared with the same concentrations of NaCl solution, serum containing Yiqi Huayu Bushen Recipe at 5%, 10% and 20% promoted cell proliferation and expression of aggrecan mRNA and decreased expression of type Ⅹ collagen mRNA (P<0.01). 10% serum containing Yiqi Huayu Bushen recipe was more effective than 5% and 20%.

Conclusion: Yiqi Huayu Bushen Recipe inhibits degeneration of the intervertebral disc cells by regulating collagen and proteoglycan expressions.

Key words: cervical spondylosis, drugs,Chinese herbal, intervertebral disk, aggrecan, collagen type Ⅹ

Ju-bao Li, Hong-yun Zhao, Yong-jian Zhao, Quan Zhou, Ning Li, Peng Xu, Qi Shi, Yong-jun Wang. Effects of Chinese herbal medicine Yiqi Huayu Bushen Recipe on expressions of aggrecan and type Ⅹ collagen mRNAs in cells from degenerated human intervertebral discs. Journal of Chinese Integrative Medicine, 2011, 9(10): 1138-1144.

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URL: http://www.jcimjournal.com/EN/10.3736/jcim20111016

http://www.jcimjournal.com/EN/Y2011/V9/I10/1138

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References 17

[1]	Gruber HE, Stasky AA, Hanley EN Jr . Characterization and phenotypic stability of human disc cells in vitro[J]. Matrix Biol, 1997,16(5):285-288 doi: 10.1016/S0945-053X(97)90016-0 pmid: 9501328
[2]	Gruber HE, Fisher EC Jr, Desai B, Stasky AA, Hoelscher G, Hanley EN Jr . Human intervertebral disc cells from the annulus: three-dimensional culture in agarose or alginate and responsiveness to TGF-beta1[J]. Exp Cell Res, 1997,235(1):13-21 doi: 10.1006/excr.1997.3647
[3]	Ma K, Hu YG, Qi ZH . Cell culture of human degenerative intervertebral disc[J]. Zhongguo Zu Zhi Gong Cheng Yan Jiu Yu Lin Chuang Kang Fu, 2009,13(2):229-232
	马可, 胡有谷, 齐宗华 . 退行性变椎间盘的细胞培养[J]. 中国组织工程研究与临床康复, 2009,13(2):229-232
[4]	Zhou Q, Wang YJ, Shi Q, Hou BX, Mo W, Hu ZJ, Liu M, Zhou CJ . Effects of “Yiqihuayu recipe” on intervertebral disc cell apoptosis in rats[J]. Shanghai Zhong Yi Yao Za Zhi, 2005,39(4):50-52 doi: 10.3969/j.issn.1007-1334.2005.04.022
	周泉, 王拥军, 施杞, 侯宝兴, 莫文, 胡志俊, 刘梅, 周重建 . 益气化瘀方对椎间盘纤维环细胞凋亡的影响[J]. 上海中医药杂志, 2005,39(4):50-52 doi: 10.3969/j.issn.1007-1334.2005.04.022
[5]	Shi Q, Wang YJ, Zhou CJ, Liu M, Zhou Q, Li CG, Sun P, Shi JX, Hou BX . Application and development of qi and blood theory in extending and relieving disc degeneration[J]. Zhongguo Zhong Yi Gu Shang Ke Za Zhi, 2006,14(S):190-192
	施杞, 王拥军, 周重建, 刘梅, 周泉, 李晨光, 孙鹏, 石继祥, 侯宝兴 . 气血理论在延缓椎间盘退变过程的运用与发展[J]. 中国中医骨伤科杂志, 2006,14(增刊):190-192
[6]	Zhou Q, Wang YJ, Shi Q, Sun P, Zhou CJ, Hu ZJ, Liu M . Effects of Yiqihuayu recipe and its decomposed formulas on apoptosis-related factors of annulus fibrosus cells in rats[J]. J Chin Integr Med, 2005,3(6):466-469
	周泉, 王拥军, 施杞, 孙鹏, 周重建, 胡志俊, 刘梅 . 益气化瘀方及其拆方对大鼠椎间盘纤维环细胞凋亡相关因子的作用[J]. 中西医结合学报, 2005,3(6):466-469
[7]	Shi JX, Wang YJ, Shi Q, Zhou Q, Li CG, Liu M, Zhou CJ . Effects on proliferation of chondrocytes derived from intervertebral disc by using Yiqihuayubushentongluo herbs with inhibitors[J]. Zhongguo Zhong Yi Gu Shang Ke Za Zhi, 2007,15(4):23-25
	石继祥, 王拥军, 施杞, 周泉, 李晨光, 刘梅, 周重建 . 益气化瘀补肾通络中药联合抑制剂对椎间盘软骨细胞增殖率的影响[J]. 中国中医骨伤科杂志, 2007,15(4):23-25
[8]	Shi Q, Wang YJ, Li CG, Zhou Q, Hu ZJ, Liu M, Zhou CJ . Study on Yiqihuayubushen recipe and its disassembled recipes in regulating mRNA expression of collagen and metabolic enzymes in extracellular matrix of cervical disc[J]. Zhongguo Zhong Xi Yi Jie He Za Zhi, 2007,27(2):142-146
	施杞, 王拥军, 李晨光, 周泉, 胡志俊, 刘梅, 周重建 . 益气化瘀补肾方及拆方影响椎间盘细胞外基质胶原和代谢酶表达的研究[J]. 中国中西医结合杂志, 2007,27(2):142-146
[9]	Wang YJ, Shi Q, Li CG, Zhou Q, Hu ZJ, Liu M, Zhou CJ . Study of Yiqi Huayu Bushen recipe and its decomposed formulas in regulating gene expressions in degenerated cervical intervertebral discs of rats[J]. J Chin Integr Med, 2006,4(6):615-619
	王拥军, 施杞, 李晨光, 周泉, 胡志俊, 刘梅, 周重建 . 益气化瘀补肾方及其拆方调控大鼠颈椎间盘基因表达谱的研究[J]. 中西医结合学报, 2006,4(6):615-619
[10]	Kluba T, Niemeyer T, Gaissmaier C, Gründer T . Human anulus fibrosis and nucleus pulposus cells of the intervertebral disc: effect of degeneration and culture system on cell phenotype[J]. Spine (Phila Pa 1976), 2005,30(24):2743-2748 doi: 10.1097/01.brs.0000192204.89160.6d
[11]	Poiraudeau S, Monteiro I, Anract P, Blanchard O, Revel M, Corvol MT . Phenotypic characteristics of rabbit intervertebral disc cells. Comparison with cartilage cells from the same animals[J]. Spine (Phila Pa 1976), 1999,24(9):837-844 doi: 10.1097/00007632-199905010-00002
[12]	Hayes AJ, Benjamin M, Ralphs JR . Extracellular matrix in development of the intervertebral disc[J]. Matrix Biol, 2001,20(2):107-121 doi: 10.1016/S0945-053X(01)00125-1 pmid: 11334712
[13]	Cs-Szabo G , Ragasa-San Juan D, Turumella V, Masuda K, Thonar EJ, An HS .Changes in mRNA and protein levels of proteoglycans of the anulus fibrosus and nucleus pulposus during intervertebral disc degeneration[J]. Spine (Phila Pa 1976), 2002,27(20):2212-2219 doi: 10.1097/00007632-200210150-00006
[14]	Roughley PJ, Alini M, Antoniou J . The role of proteoglycans in aging, degeneration and repair of the intervertebral disc[J]. Biochem Soc Trans, 2002,30(Pt 6):869-874 doi: 10.1042/bst0300869
[15]	He HL, Wu ZH, Zhang JG, Wang YP, Zhou Y, Xu YQ, Yuan JG, Qiu GX . Primary study on collagen Ⅹ gene expression in the apical disc of idiopathic scoliosis[J]. Zhonghua Yi Xue Za Zhi, 2004,84(20):1681-1686
	何海龙, 吴志宏, 仉建国, 王以朋, 周严, 徐雅琴, 袁建刚, 邱贵兴 . Ⅹ型胶原基因在特发性脊柱侧凸患者顶椎椎间盘内表达的初步研究[J]. 中华医学杂志, 2004,84(20):1681-1686
[16]	Xi YM, Hu YG, Lü ZH, Zheng HJ, Chen Y, Qi Z . Gene expression of collagen types Ⅸ and Ⅹ in the lumbar disc[J]. Chin J Traumatol, 2004,7(2):76-80
[17]	Nerlich AG, Schleicher ED, Boos N . 1997 Volvo Award winner in basic science studies. Immunohistologic markers for age-related changes of human lumbar intervertebral discs[J]. Spine (Phila Pa 1976), 1997,22(24):2781-2795 doi: 10.1097/00007632-199712150-00001

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Gene	Forward primer 5′-3′	Reverse primer 5′-3′	Accession number	Product length (bp)
β-actin	AAGGTGACAGCAGTCGGTT	TGTGTGGACTTGGGAGAGG	NM_177423	195
Aggrecan	GCCAGCACCACCAATGTAAG	TTCAGTAACACCCTCCACGAAC	NM_036445	116
Collagen 10α1	AGTGTTTTACGCTGAACG	TGCTCTCCTCTTACTGCT	NM_000493	123

	Content volume (μL)
Master mix	10
RNase-free water	7
Forward primer	1
Reverse primer	1
cDNA	1
Total volume	20

Group	Optical density value
Group	24 h	48 h
5% saline serum	0.42±0.03	0.67±0.09
10% saline serum	0.71±0.18	0.84±0.21
20% saline serum	1.05±0.34	1.13±0.27
5% Yiqi Huayu Bushen recipe serum	1.68±0.31^**△	1.92±0.36^**△
10% Yiqi Huayu Bushen recipe serum	2.17±0.25^**	2.39±0.44^**
20% Yiqi Huayu Bushen recipe serum	1.53±0.23^**△	1.64±0.49^**△

Group	n	Aggrecan mRNA
Group	n	5 h	48 h	72 h
5% saline serum	6	1.03±0.05	1.06±0.09	1.04±0.03
10% saline serum	6	1.12±0.07	1.27±0.09	1.13±0.05
20% saline serum	6	1.26±0.04	1.23±0.27	1.39±0.12
5% Yiqi Huayu Bushen recipe serum	6	3.08±0.86^**△	5.39±1.23^**△	3.57±1.15^**△
10% Yiqi Huayu Bushen recipe serum	6	6.33±1.12^**	9.50±1.87^**	7.63±1.42^**
20% Yiqi Huayu Bushen recipe serum	6	3.25±0.99^**△	6.28±1.96^**△	4.05±0.34^**△

Group	n	Type Ⅹ collagen mRNA
Group	n	5 h	48 h	72 h
5% saline serum	6	1.02±0.03	1.01±0.02	1.03±0.06
10% saline serum	6	1.01±0.03	1.03±0.02	0.98±0.16
20% saline serum	6	1.03±0.05	1.05±0.04	0.96±0.22
5% Yiqi Huayu Bushen recipe serum	6	0.74±0.14^**△	0.55±0.17^**△	0.73±0.09^**△
10% Yiqi Huayu Bushen recipe serum	6	0.48±0.11^**	0.21±0.03^**	0.35±0.16^**
20% Yiqi Huayu Bushen recipe serum	6	0.63±0.16^**△	0.38±0.05^**△	0.64±0.13^**△

Effects of Chinese herbal medicine Yiqi Huayu Bushen Recipe on expressions of aggrecan and type Ⅹ collagen mRNAs in cells from degenerated human intervertebral discs

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