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[1]夏炳江,侯明生,張磊,等.五福飲對大鼠尾部退變椎間盤的影響及其作用機(jī)制…[J].中醫(yī)正骨,2018,30(01):18-24.
　XIA Bingjiang,HOU Mingsheng,ZHANG Lei,et al.Effects of Wufu Yin(五福飲)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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五福飲對大鼠尾部退變椎間盤的影響及其作用機(jī)制…()
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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第30卷
期數(shù):
2018年01期
頁碼:
18-24
欄目:
基礎(chǔ)研究
出版日期:
2018-01-20

文章信息/Info

Title:
Effects of Wufu Yin(五福飲)on caudal degenerative intervertebral disc in rats and its mechanism of action
作者:
夏炳江1; 侯明生1; 張磊2; 韋金忠1; 胡松峰1
1.浙江省紹興市中醫(yī)院,浙江 紹興 312000; 2.浙江中醫(yī)藥大學(xué),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
關(guān)鍵詞:
椎間盤退行性變;  五福飲;  β連環(huán)素;  蛋白聚糖類;  含Ⅰ型血小板結(jié)合蛋白基序的解聚蛋白樣金屬蛋白酶;  大鼠;  動物實(shí)驗(yàn)
Keywords:
Keywords intervertebral disc degeneration;  Wufu Yin;  beta Catenin;  proteoglycans;  a disintesrin and metalloprotease with thrombospondin typeⅠmotifs;  rats;  animal experimentation
摘要:
目的:觀察五福飲對大鼠尾部退變椎間盤的影響及其作用機(jī)制。方法:將48只12周齡健康雄性清潔級SD大鼠隨機(jī)分成空白對照組、模型組、五福飲組,每組16只。針刺模型組、五福飲組大鼠尾部椎間盤并向椎間盤內(nèi)注入腫瘤壞死因子-α,空白對照組大鼠不做任何處理。造模成功后(造模開始后4周)五福飲組以20 mL·kg-1劑量的五福飲灌胃,空白對照組和模型組給予等量生理鹽水灌胃,每日1次,連續(xù)灌胃8周。分別于造模成功后及藥物干預(yù)8周后從各組隨機(jī)抽取8只大鼠,采用麻醉過量法處死后,完整取下Co5～6椎間盤及其相鄰兩側(cè)椎體,行Micro-CT掃描測量椎間相對高度,采取HE染色觀察椎間盤組織形態(tài),采用逆轉(zhuǎn)錄-聚合酶鏈?zhǔn)椒磻?yīng)法檢測椎間盤β-連環(huán)蛋白(β-catenin)和含Ⅰ型血小板結(jié)合蛋白基序的解聚蛋白樣金屬蛋白酶(a disintesrin and metalloprotease with thrombospondin typeⅠmotifs,ADAMTS)-5基因的表達(dá)。結(jié)果:①椎間隙相對高度。造模成功后,3組大鼠Co5～Co6椎間隙相對高度比較,差異有統(tǒng)計學(xué)意義[100%,(56.37±11.32)%,(55.63±12.76)%,F=22.148,P=0.000]; 空白對照組Co5～Co6椎間隙相對高度高于模型組和五福飲組(P=0.000,P=0.000); 模型組Co5～Co6椎間隙相對高度與五福飲組比較,差異無統(tǒng)計學(xué)意義(P=0.203)。藥物干預(yù)8周后,3組大鼠Co5～Co6椎間隙相對高度比較,差異有統(tǒng)計學(xué)意義[100%,(50.35±10.72)%,(80.84±9.43)%,F=31.492,P=0.000]; 空白對照組Co5～Co6椎間隙相對高度高于模型組和五福飲組(P=0.000,P=0.000),模型組Co5～Co6椎間隙相對高度低于五福飲組(P=0.001)。②椎間盤組織形態(tài)。造模成功后,模型組和五福飲組大鼠Co5～6椎間盤均明顯退變,表現(xiàn)為髓核細(xì)胞數(shù)量減少,纖維環(huán)排列紊亂且出現(xiàn)裂隙,纖維環(huán)與髓核交界區(qū)變窄。藥物干預(yù)8周后,模型組大鼠Co5～6椎間盤退變更明顯,而五福飲組大鼠Co5～6椎間盤退變情況明顯改善,可見較多髓核細(xì)胞,細(xì)胞排列尚均勻,纖維環(huán)膠原走向清楚,纖維環(huán)與髓核交界區(qū)較寬。③椎間盤組織β-catenin基因的表達(dá)。造模成功后,3組大鼠Co5～6椎間盤組織β-catenin基因表達(dá)量比較,差異有統(tǒng)計學(xué)意義(1.32±0.21,2.87±0.41,2.73±0.42,F=18.806,P=0.001); 空白對照組Co5～6椎間盤組織β-catenin基因表達(dá)量低于模型組和五福飲組(P=0.000,P=0.001); 模型組Co5～6椎間盤組織β-catenin基因表達(dá)量與五福飲組比較,差異無統(tǒng)計學(xué)意義(P=0.457)。藥物干預(yù)8周后,3組大鼠Co5～6椎間盤組織β-catenin基因表達(dá)量比較,差異有統(tǒng)計學(xué)意義(1.01±0.17,2.64±0.33,1.51±0.31,F=26.616,P=0.000); 空白對照組Co5～6椎間盤組織β-catenin基因表達(dá)量低于模型組和五福飲組(P=0.000,P=0.002),模型組Co5～6椎間盤組織β-catenin基因表達(dá)量高于五福飲組(P=0.001)。④椎間盤組織ADAMTS-5基因的表達(dá)。造模成功后,3組大鼠Co5～6椎間盤組織ADAMTS-5基因表達(dá)量比較,差異有統(tǒng)計學(xué)意義(1.18±0.14,2.34±0.25,2.42±0.28,F=47.786,P=0.000); 空白對照組Co5～6椎間盤組織ADAMTS-5基因表達(dá)量低于模型組和五福飲組(P=0.000,P=0.000); 模型組Co5～6椎間盤組織ADAMTS-5基因表達(dá)量與五福飲組比較,差異無統(tǒng)計學(xué)意義(P=0.530)。藥物干預(yù)8周后,3組大鼠Co5～6椎間盤組織ADAMTS-5基因表達(dá)量比較,差異有統(tǒng)計學(xué)意義(1.21±0.27,2.45±0.29,1.38±0.13,F=6.053,P=0.022); 空白對照組和五福飲組Co5～6椎間盤組織ADAMTS-5基因表達(dá)量低于模型組(P=0.000,P=0.027); 空白對照組Co5～6椎間盤組織ADAMTS-5基因表達(dá)量與五福飲組比較,差異無統(tǒng)計學(xué)意義(P=0.205)。結(jié)論:五福飲可緩解大鼠尾部椎間盤組織退變,恢復(fù)椎間隙相對高度,其作用機(jī)制可能與其能下調(diào)椎間盤組織中β-catenin和ADAMTS-5的表達(dá)有關(guān)。
Abstract:
ABSTRACT Objective:To study the effects of Wufu Yin(五福飲,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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備注/Memo

備注/Memo:
基金項(xiàng)目:浙江省中醫(yī)藥科技計劃項(xiàng)目(2017ZB091); 浙江省紹興市科技計劃項(xiàng)目(2015B70060) 通訊作者:夏炳江 E-mail:[email protected]
更新日期/Last Update: 2018-06-02