[1]邓蓉蓉,王彦超,席志鹏,等.“加味三痹汤”对大鼠骨髓间充质干细胞归巢及对椎间盘退变的影响[J].中国中医骨伤科杂志,2019,27(05):14-18.
　DENG Rongrong,WANG Yanchao,XI Zhipeng,et al.Observing the Effect of Jiawei Sanbi Decoction on Homing of Rat Bone Mesenchymal Stem Cells and Inhibiting Intervertebral Disc Degeneration[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2019,27(05):14-18.
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“加味三痹汤”对大鼠骨髓间充质干细胞归巢及对椎间盘退变的影响()
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《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第27卷
期数:
2019年05期
页码:
14-18
栏目:
实验研究
出版日期:
2019-05-15

文章信息/Info

Title:
Observing the Effect of Jiawei Sanbi Decoction on Homing of Rat Bone Mesenchymal Stem Cells and Inhibiting Intervertebral Disc Degeneration
文章编号:
1005-0205(2019)05-0014-05
作者:
邓蓉蓉1; 王彦超2; 席志鹏1; 康然1; 强东坤1; 顾军1; 谢林1△
1江苏省中西医结合医院,骨伤科 2河南省人民医院
Author(s):
DENG Rongrong1; WANG Yanchao2; XI Zhipeng1; KANG Ran1;  QIANG Dongkun1; GU Jun1; XIE Lin1△
1Jiangsu Province Hospital on Intergration of Chinese and Western Medicine,Nanjing 210028,China; 2Henan Province People's Hospital,Zhengzhou 450000,China.
关键词:
加味三痹汤;  骨髓间充质干细胞;  归巢;  椎间盘退行性疾病
Keywords:
Jiawei Sanbi decoction;  rat bone mesenchymal stem cells;  homing;  intervertebral disc degeneration diseases
分类号:
R-33
文献标志码:
A
摘要:
目的:体内实验观察“加味三痹汤”能否促进外源性大鼠骨髓间充质干细胞(rBMSCs)迁移、归巢至退变的椎间盘,从而探索椎间盘退变修复的可能机制。方法:贴壁法提取rBMSC,采用PKH26红色荧光标记试剂盒标记细胞。针刺法构建大鼠尾椎间盘退变模型,根据改良的Thompson分级标准对退变模型进行分级,选取退变Ⅲ级大鼠随机分组进行后续体内实验。将标记PKH26的rBMSCs通过尾静脉注射移植实验大鼠体内,小动物活体成像技术观察中药灌服后rBMSCs迁移和归巢行为; MRI观察中药对椎间盘退变的作用; 免疫组化染色观察中药对椎间盘蛋白多糖及Ⅱ型胶原含量的影响。结果:早期中药组对BMSCs迁移和归巢至退变椎间盘的数量未见明显影响,但是在中后期,中药组能够促进移植的干细胞归巢并维持其数量,同时中药组提高退变椎间盘组织中蛋白多糖和Ⅱ型胶原的含量,MRI观察中药抑制椎间盘退变。结论:“加味三痹汤”能够促进干细胞归巢和抑制椎间盘退变,对于干细胞移植联合中医药治疗具有一定指导意义,在再生医学应用于椎间盘退变治疗方面具有一定的前景。
Abstract:
Objective:To observe whether Jiawei Sanbi decoction can promote the migration of exogenous rat bone mesenchymal stem cells(rBMSCs)homing to intervertebral disc and inhibit the degeneration in vivo, so as to explore the possible mechanism in repair of intervertebral disc degeneration.Methods:Rat BMSCs extracted by cell attachment method were labeled by PKH26 and identified by the cell morphology and flow cytometry. SD rats were punctured with 18G needles in Co7/8 and Co9/10 intervertebral discs, MRI was used to assess the degeneration grade according to the improved Thompson grading standard. Rats of grade Ⅲ were randomly divided into four groups for vivo experiment. Rats of the experimental group were transplanted with BMSCs by tail vein injection, and the migration of BMSCs was observed by the small animal imaging techniques in vivo. MRI was used to assess the degeneration grade and immunohistochemical staining(IHC)was used to detect the content of proteoglycan and collagen Ⅱ.Results:Chinese medicine had no significant effect on BMSCs migration and homing comparing with the control group in the first week, which happens in the 2 and 4 weeks. At the same time, more proteoglycan and collagen Ⅱ were detected and the inhibition of degeneration in Chinese medicine group.Conclusion:Jiawei Sanbi decoction have advantages of BMSCs homing and inhibiting the degeneration of intervertebral disc, which suggest stem cell transplantation combined with traditional Chinese medicine treatment can be applicated in regenerating the degeneration of intervertebral disc.

参考文献/References:

[1] LOZANO R,NAGHAVI M,FOREMAN K,et al.Global and regional mortality from 235 causes of death for 20 age groups in 1990 and 2010:a systematic analysis for the global burden of disease study 2010[J].Lancet,2012,380(9859):2095-2128.
[2] RISBUD M V,SHAPIRO I M.Role of cytokines in intervertebral disc degeneration:pain and disc content[J].Nat Rev Rheumatol,2014,10(1):44-56.
[3] ILLIEN-JüNGER S,PATTAPPA G,PEROGLIO M,et al.Homing of mesenchymal stem cells in induced degenerative intervertebral discs in a whole organ culture system[J].Spine(Phila Pa 1976),2012,37(22):1865-1873.
[4] 王彦超,邓蓉蓉,叶亚东,等.补肾活血舒筋方含药血清对大鼠骨髓间充质干细胞增殖、迁移的影响及其机制[J].山东医药,2017,5(16):38-40.
[5] 赵海滨,李晔紫,师伟,等.活血化瘀法对急性心肌梗死大鼠骨髓干细胞动员入血及定向归巢影响实验研究[J].慢性病学杂志,2015,16(3):301-304.
[6] 许敬人,詹红生.椎间盘退变机制及中药对其的干预作用[J].临床和实验医学杂志,2015,14(19):1657-1660.
[7] 强东坤,邓蓉蓉,顾军,等.脊伸痹通方联合西乐葆治疗腰椎间盘突出症临床观察[J].西部中医药,2018,31(5):90-92.
[8] 强东坤,邓蓉蓉,顾军.从气血、肝肾论治腰椎间盘突出症经验撷粹[J].江苏中医药,2018,50(2):20-21.
[9] 张仕兵,谢林,康然,等.全内窥镜下椎板间隙技术配合补肾活血舒筋方治疗腰椎间盘突出症55例疗效分析[J].实用中西医结合临床,2016,16(8):47-48.
[10] 谢林,康然,刘彬彬,等.康颈颗粒对大鼠颈椎间盘细胞凋亡的影响[J].现代中西医结合杂志,2009,18(31):3800-3801.
[11] ZARUBA,M M,THEISS H D,VALLASTER M,et al.Synergy between CD26/DPP-IV inhibition and G-CSF improves cardiac function after acute myocardial infarction[J].Cell Stem Cell,2009,4(4):313-323.
[12] WAGERS A J.The stem cell niche in regenerative medicine[J].Cell Stem Cell,2012,10(4):362-369.
[13] SAKAI D,ANDERSSON G B.Stem cell therapy for intervertebral disc regeneration:obstacles and solutions[J].Nat Rev Rheumatol,2015,11(4):243-256.
[14] GILBERT H T,HOYLAND J A,RICHARDSON S M.Stem cell regeneration of degenerated intervertebral discs:current status(update)[J].Curr Pain Headache Rep,2013,17(12):377.
[15] HUANG Z,ZHANG L,TENG X,et al.A new in vivo method to retard progression of intervertebral disc degeneration through stimulation of endogenous stem cells with simvastatin[J].Med Hypotheses,2017,32(101):65-66.
[16] 王倩,补阳还五汤对大鼠急性脑缺血后神经干细胞迁移的影响[J].山东中医杂志,2013(10):742-744.
[17] 郭占鹏,刘堃,黄米娜,等.补阳还五汤对大鼠肌源性干细胞体外生长分化的影响[J].中成药,2015,37(9):2049-2051.
[18] CHEN T,WANG J,LIU M,et al.Screening of natural compounds with neuronal differentiation promoting effects in a cell-based model[J].Chinese Journal of Natural Medicines,2015,13(8):602-608.
[19] XU Y,DAIG J,DAI G J,LIU Q,et al.Effect of ermiao srecipe with medicinal guide angelicae pubescentis radix on promoting the homing of bone marrow stem cells to treat cartilage damage in osteoarthritis rats[J].Chinese Journal of Integrative Medicine,2014,20(8):600-609.
[20] MAZAKI T,KITAJIMA T,SHZOZAKI Y,et al.In vitro and in vivo enhanced osteogenesis by kaempferol found by a high-throughput assay using human mesenchymal stromal cells[J].Journal of Functional Foods,2014,6(1):241-247.

备注/Memo

备注/Memo:
基金项目:国家自然科学基金面上项目(81772356)江苏省中医药领军人才项目(SLJ0210)江苏省干部保健科研项目(BJ16026)江苏省卫生健康委员会项目(H2018025)△通信作者 E-mail:xielin117@126.com
更新日期/Last Update: 2019-05-15