[1]夏炳江,侯明生,张磊,等.五福饮对大鼠尾部退变椎间盘的影响及其作用机制…[J].中医正骨,2018,30(01):18-24.
　XIA Bingjiang,HOU Mingsheng,ZHANG Lei,et al.Effects of Wufu Yin(五福饮)on caudal degenerative intervertebral disc in rats and its mechanism of action[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2018,30(01):18-24.
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五福饮对大鼠尾部退变椎间盘的影响及其作用机制…()
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《中医正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第30卷
期数:
2018年01期
页码:
18-24
栏目:
基础研究
出版日期:
2018-01-20

文章信息/Info

Title:
Effects of Wufu Yin(五福饮)on caudal degenerative intervertebral disc in rats and its mechanism of action
作者:
夏炳江1; 侯明生1; 张磊2; 韦金忠1; 胡松峰1
1.浙江省绍兴市中医院,浙江 绍兴 312000; 2.浙江中医药大学,浙江 杭州 310053
Author(s):
XIA Bingjiang1; HOU Mingsheng1; ZHANG Lei2; WEI Jinzhong1; HU Songfeng1
1.Shaoxing Hospital of Traditional Chinese Medicine,Shaoxing 312000,Zhejiang,China 2.Zhejiang Chinese Medical University,Hangzhou 310053,Zhejiang,China
关键词:
椎间盘退行性变;  五福饮;  β连环素;  蛋白聚糖类;  含Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶;  大鼠;  动物实验
Keywords:
Keywords intervertebral disc degeneration;  Wufu Yin;  beta Catenin;  proteoglycans;  a disintesrin and metalloprotease with thrombospondin typeⅠmotifs;  rats;  animal experimentation
摘要:
目的:观察五福饮对大鼠尾部退变椎间盘的影响及其作用机制。方法:将48只12周龄健康雄性清洁级SD大鼠随机分成空白对照组、模型组、五福饮组,每组16只。针刺模型组、五福饮组大鼠尾部椎间盘并向椎间盘内注入肿瘤坏死因子-α,空白对照组大鼠不做任何处理。造模成功后(造模开始后4周)五福饮组以20 mL·kg-1剂量的五福饮灌胃,空白对照组和模型组给予等量生理盐水灌胃,每日1次,连续灌胃8周。分别于造模成功后及药物干预8周后从各组随机抽取8只大鼠,采用麻醉过量法处死后,完整取下Co5～6椎间盘及其相邻两侧椎体,行Micro-CT扫描测量椎间相对高度,采取HE染色观察椎间盘组织形态,采用逆转录-聚合酶链式反应法检测椎间盘β-连环蛋白(β-catenin)和含Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶(a disintesrin and metalloprotease with thrombospondin typeⅠmotifs,ADAMTS)-5基因的表达。结果:①椎间隙相对高度。造模成功后,3组大鼠Co5～Co6椎间隙相对高度比较,差异有统计学意义[100%,(56.37±11.32)%,(55.63±12.76)%,F=22.148,P=0.000]; 空白对照组Co5～Co6椎间隙相对高度高于模型组和五福饮组(P=0.000,P=0.000); 模型组Co5～Co6椎间隙相对高度与五福饮组比较,差异无统计学意义(P=0.203)。药物干预8周后,3组大鼠Co5～Co6椎间隙相对高度比较,差异有统计学意义[100%,(50.35±10.72)%,(80.84±9.43)%,F=31.492,P=0.000]; 空白对照组Co5～Co6椎间隙相对高度高于模型组和五福饮组(P=0.000,P=0.000),模型组Co5～Co6椎间隙相对高度低于五福饮组(P=0.001)。②椎间盘组织形态。造模成功后,模型组和五福饮组大鼠Co5～6椎间盘均明显退变,表现为髓核细胞数量减少,纤维环排列紊乱且出现裂隙,纤维环与髓核交界区变窄。药物干预8周后,模型组大鼠Co5～6椎间盘退变更明显,而五福饮组大鼠Co5～6椎间盘退变情况明显改善,可见较多髓核细胞,细胞排列尚均匀,纤维环胶原走向清楚,纤维环与髓核交界区较宽。③椎间盘组织β-catenin基因的表达。造模成功后,3组大鼠Co5～6椎间盘组织β-catenin基因表达量比较,差异有统计学意义(1.32±0.21,2.87±0.41,2.73±0.42,F=18.806,P=0.001); 空白对照组Co5～6椎间盘组织β-catenin基因表达量低于模型组和五福饮组(P=0.000,P=0.001); 模型组Co5～6椎间盘组织β-catenin基因表达量与五福饮组比较,差异无统计学意义(P=0.457)。药物干预8周后,3组大鼠Co5～6椎间盘组织β-catenin基因表达量比较,差异有统计学意义(1.01±0.17,2.64±0.33,1.51±0.31,F=26.616,P=0.000); 空白对照组Co5～6椎间盘组织β-catenin基因表达量低于模型组和五福饮组(P=0.000,P=0.002),模型组Co5～6椎间盘组织β-catenin基因表达量高于五福饮组(P=0.001)。④椎间盘组织ADAMTS-5基因的表达。造模成功后,3组大鼠Co5～6椎间盘组织ADAMTS-5基因表达量比较,差异有统计学意义(1.18±0.14,2.34±0.25,2.42±0.28,F=47.786,P=0.000); 空白对照组Co5～6椎间盘组织ADAMTS-5基因表达量低于模型组和五福饮组(P=0.000,P=0.000); 模型组Co5～6椎间盘组织ADAMTS-5基因表达量与五福饮组比较,差异无统计学意义(P=0.530)。药物干预8周后,3组大鼠Co5～6椎间盘组织ADAMTS-5基因表达量比较,差异有统计学意义(1.21±0.27,2.45±0.29,1.38±0.13,F=6.053,P=0.022); 空白对照组和五福饮组Co5～6椎间盘组织ADAMTS-5基因表达量低于模型组(P=0.000,P=0.027); 空白对照组Co5～6椎间盘组织ADAMTS-5基因表达量与五福饮组比较,差异无统计学意义(P=0.205)。结论:五福饮可缓解大鼠尾部椎间盘组织退变,恢复椎间隙相对高度,其作用机制可能与其能下调椎间盘组织中β-catenin和ADAMTS-5的表达有关。
Abstract:
ABSTRACT Objective:To study the effects of Wufu Yin(五福饮,WFY)on caudal degenerative intervertebral disc in rats and its mechanism of action.Methods:Forty-eight 12-week-old healthy clean-grade male SD rats were randomly divided into blank control group,model group and WFY group,16 cases in each group.The tumor necrosis factor-α(TNF-α)were injected into caudal intervertebral disc of rats in model group and WFY group,while the rats in blank control group were not given any surgical intervention.After successful modeling,the rats in WFY group were intragastric administrated with WFY(20 mL/kg),while the rats in blank control group and model group were intragastric administrated with same dose of normal saline,once a day for 8 consecutive weeks.Eight rats were randomly selected from each group and were executed by overdose anesthesia after successful modeling and 8-weeks drug intervention respectively.Their Co5-6 intervertebral discs and bilateral adjacent vertebral bodies were fetched out completely for measuring the intervertebral relative height by Micro-CT scanning.The intervertebral disc tissue morphology was observed through HE staining and the genetic expressions of β-catenin and a disintesrin and metalloprotease with thrombospondin typeⅠmotifs(ADAMTS)-5 in intervertebral discs were detected by using reverse transcription-polymerase chain reaction(RT-PCR)assays.Results:After successful modeling,there was statistical difference in the relative height of Co5/Co6 intervertebral space between the 3 groups(100%,56.37+/-11.32%,55.63+/-12.76%,F=22.148,P=0.000).The relative height of Co5/Co6 intervertebral space was higher in blank control group compared to model group and WFY group(P=0.000,P=0.000),and there was no statistical difference in the relative height of Co5/Co6 intervertebral space between model group and WFY group(P=0.203).After 8-week drug intervention,there was statistical difference in the relative height of Co5/Co6 intervertebral space between the 3 groups(100%,50.35+/-10.72%,80.84+/-9.43%,F=31.492,P=0.000).The relative height of Co5/Co6 intervertebral space was higher in blank control group compared to model group and WFY group(P=0.000,P=0.000)and was lower in model group compared to WFY group(P=0.001).After successful modeling,the Co5/Co6 intervertebral discs of rats in model group and WFY group degenerated significantly.The degeneration presented with decreased nucleus pulposus cells,disorderly and cracked fibrous ring and narrow junctional zone of fibrous ring and nucleus pulposus.After 8-week drug intervention,more obvious degeneration of Co5/Co6 intervertebral discs of rats was found in model group; while the degeneration of Co5/Co6 intervertebral discs was significantly improved in rats of WFY group,presented with increasing nucleus pulposus cells arranged in order,clear fibrous ring collagen and wide junctional zone of fibrous ring and nucleus pulposus.After successful modeling,there was statistical difference in genetic expressions of β-catenin in Co5/Co6 intervertebral disc tissues between the 3 groups(1.32+/-0.21,2.87+/-0.41,2.73+/-0.42,F=18.806,P=0.001).The genetic expressions of β-catenin in Co5/Co6 intervertebral disc tissues were lower in blank control group compared to model group and WFY group(P=0.000,P=0.001),and there was no statistical difference in genetic expressions of β-catenin in Co5/Co6 intervertebral disc tissues between model group and WFY group(P=0.457).After 8-week drug intervention,there was statistical difference in genetic expressions of β-catenin in Co5/Co6 intervertebral disc tissues between the 3 groups(1.01+/-0.17,2.64+/-0.33,1.51+/-0.31,F=26.616,P=0.000).The genetic expressions of β-catenin in Co5/Co6 intervertebral disc tissues were lower in blank control group compared to model group and WFY group(P=0.000,P=0.002),and were higher in model group compared to WFY group(P=0.001).After successful modeling,there was statistical difference in genetic expressions of ADAMTS-5 in Co5/Co6 intervertebral disc tissues between the 3 groups(1.18+/-0.14,2.34+/-0.25,2.42+/-0.28,F=47.786,P=0.000).The genetic expressions of ADAMTS-5 in Co5/Co6 intervertebral disc tissues were lower in blank control group compared to model group and WFY group(P=0.000,P=0.000),and there was no statistical difference in genetic expressions of ADAMTS-5 in Co5/Co6 intervertebral disc tissues between model group and WFY group(P=0.530).After 8-week drug intervention,there was statistical difference in genetic expressions of ADAMTS-5 in Co5/Co6 intervertebral disc tissues between the 3 groups(1.21+/-0.27,2.45+/-0.29,1.38+/-0.13,F=6.053,P=0.022).The genetic expressions of ADAMTS-5 in Co5/Co6 intervertebral disc tissues were lower in blank control group and WFY group compared to model group(P=0.000,P=0.027),and there was no statistical difference in genetic expressions of ADAMTS-5 in Co5/Co6 intervertebral disc tissues between blank control group and WFY group(P=0.205).Conclusion:WFY can relieve the degeneration of rat's caudal intervertebral disc tissue and restore the relative height of intervertebral space,and its mechanisms of action may be that it can down-regulate the expression of β-catenin and ADAMTS-5 in intervertebral disc tissues.

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备注/Memo

备注/Memo:
基金项目:浙江省中医药科技计划项目(2017ZB091); 浙江省绍兴市科技计划项目(2015B70060) 通讯作者:夏炳江 E-mail:xiabj2006@163.com
更新日期/Last Update: 2018-06-02