[1]杨光露 郭杨△ 马勇 涂鹏程 孙杰 许炜民 吴承杰.扶阳宣痹汤对大鼠椎间盘退变及基质金属蛋白酶表达的影响[J].中国中医骨伤科杂志,2021,29(04):1-7.
　YANG Guanglu GUO Yang MA Yong TU PengchengSUN Jie XU Weimin WU Chengjie.Effect of Fuyang Xuanbi Decoction on Intervertebral Disc Degenerationand the Expression of Matrix Metalloproteinase in Rats[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2021,29(04):1-7.
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扶阳宣痹汤对大鼠椎间盘退变及基质金属蛋白酶表达的影响()
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《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第29卷
期数:
2021年04期
页码:
1-7
栏目:
实验研究
出版日期:
2021-04-15

文章信息/Info

Title:
Effect of Fuyang Xuanbi Decoction on Intervertebral Disc Degenerationand the Expression of Matrix Metalloproteinase in Rats
文章编号:
1005-0205(2021)04-0001-07
作者:
杨光露1; 2 郭杨1; 2△ 马勇1; 2 涂鹏程1; 2 孙杰1; 2 许炜民1; 2 吴承杰1; 2
1南京中医药大学附属医院骨伤科(南京,210029)
2南京中医药大学骨伤修复与重建新技术实验室
Author(s):
YANG Guanglu1; 2 GUO Yang1; 2△ MA Yong1; 2 TU Pengcheng1; 2SUN Jie1; 2 XU Weimin1; 2 WU Chengjie1; 2
1Department of Traumatology & Orthopedics,Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China; 2Laboratory of New Techniques of Restoration & Reconstruction of Orthopedics and Traumatology,Nanjing University of Chinese Medicine,Nanjing 210023,China.
关键词:
扶阳宣痹汤;  椎间盘退变;  基质金属蛋白酶9;  基质金属蛋白酶13;  髓核基质
Keywords:
fuyang xuanbi decoction;  intervertebral disc degeneration;  MMP9;  MMP13;  nucleus pulposus matrix
分类号:
R-33
文献标志码:
A
摘要:
目的:观察扶阳宣痹汤对退变椎间盘组织病理及相关因子MMP9和MMP13表达的影响,探讨改善椎间盘退变的作用机制。方法:选择大鼠尾椎间盘穿刺Co5/6/7建立椎间盘退变模型,随机分为空白组、模型组、腰痹通组(腰痹通0.34 g/(kg·d))、扶阳宣痹汤低剂量组(8.19 g/(kg·d))、中剂量组(16.38 g/(kg·d))、高剂量组(32.76 g/(kg·d)),制备大鼠椎间盘退变模型,模型制备成功后对各组大鼠进行灌胃给药,给药4周后行磁共振成像(MRI)复查各组大鼠尾椎间盘退变情况,处死大鼠提取尾椎间盘髓核组织,使用苏木精-伊红(HE)染色检测椎间盘退变程度,免疫组化检测椎间盘组织COL2A1和Aggrecan的表达,蛋白免疫印迹法(Western Blot)检测椎间盘组织MMP9和MMP13的蛋白表达,实时聚合酶链式反应(RT-PCR)检测椎间盘组织COL2A1、Aggrecan、MMP9和MMP13的mRNA表达。结果:术后4周与模型组相比,扶阳宣痹汤低、中、高剂量组椎间盘T2加权信号强度高,高剂量组MRI的Piffimann椎间盘评分明显降低,差异有统计学意义(P<0.01); HE染色的Masuda评分明显降低,差异有统计学意义(P<0.05); 对椎间盘COL2A1和Aggrecan的免疫组化染色更强,差异有统计学意义(P<0.01); 显著下调MMP9和MMP13的mRNA表达及蛋白表达,差异有统计学意义(P<0.05,P<0.01); 同时上调COL2A1和Aggrecan的mRNA表达,差异有统计学意义(P<0.05)。结论:扶阳宣痹汤可以通过抑制MMP9和MMP13表达促进COL2A1及Aggrecan的合成,改善椎间盘退行性病变,并存在一定剂量依赖性。本研究为扶阳宣痹汤改善椎间盘退变提供了实验依据。
Abstract:
To observe the effect of Fuyang Xuanbi decoction on histopathology of degenerative intervertebral disc and expression of related factors MMP9 and MMP13,and to explore the mechanism of improving intervertebral disc degeneration.Methods:The rat model of intervertebral disc degeneration was established by Co5/6/7 puncture of caudal intervertebral disc.The rats were randomly divided into blank group,model group,Yaobitong group(Yaobitong 0.34 g/(kg·d)),low dose group(Fuyang Xuanbi decoction 8.19 g/(kg·d)),middle dose group(Fuyang Xuanbi decoction 16.38(g/kg·d))and high dose group(Fuyang Xuanbi decoction 32.76 g/(kg·d)).After all the models were successfully established,rats in each group were given intragastric administration of the drug.Magnetic resonance imaging(MRI)was performed to reexamine the degeneration of rats caudal intervertebral disc after 4 weeks of administration,and rats were sacrificed to extract the nucleus pulposus of caudal intervertebral disc.The degree of intervertebral disc degeneration was detected by Hematoxylin eosin(HE)staining,the expression of COL2A1 and aggrecan in intervertebral disc tissue was detected by immunohistochemistry,and the protein expression of MMP9 and MMP13 were detected by Western Blot.Realtime-polymerase chain reaction(RT-PCR)was used to detect the mRNA expression of COL2A1,aggrecan,MMP9 and MMP13 in intervertebral disc tissue.Results:The T2-weighted signal intensities of intervertebral disc in the low,middle and high dose groups of Fuyang Xuanbi decoction were higher than that in the model group at 4 weeks after operation.Additionally,the piffimann intervertebral disc score of MRI and the Masuda score of HE staining in the high dose group were significantly lower than those in the model group,but the immunohistochemical staining of COL2A1 and Aggrecan of the intervertebral disc were stronger.The mRNA expression and protein expression of MMP9 and MMP13 were significantly down-regulated,while the mRNA expression of COL2A1 and Aggrecan was up-regulated.Conclusion:Fuyang Xuanbi decoction can promote the synthesis of COL2A1 and MMP13 by inhibiting the expression of COL2A1 and MMP13,and improve intervertebral disc degeneration in a dose-dependent manner.This study provides some experimental basis for Fuyang Xuanbi decoction to improve intervertebral disc degeneration.

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备注/Memo

备注/Memo:
基金项目:江苏省南京市研究生教育创新计划资助项目立项课题(SJCX18_0570)
△通信作者 E-mail:drguoyang@126.com
更新日期/Last Update: 2021-04-15