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[1]趙赫,高譽(yù)珊,胡振國(guó),等.補(bǔ)腎壯筋湯對(duì)腰椎間盤(pán)退變大鼠椎間盤(pán)及行為學(xué)特征影響的實(shí)驗(yàn)研究[J].中醫(yī)正骨,2017,29(07):12-20,26.
　ZHAO He,GAO Yushan,HU Zhenguo,et al.Effect of Bushen Zhuangjin Tang(補(bǔ)腎壯筋湯)on intervertebral disc and behavioral characteristics of rats with lumbar intervertebral disc degeneration[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2017,29(07):12-20,26.
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補(bǔ)腎壯筋湯對(duì)腰椎間盤(pán)退變大鼠椎間盤(pán)及行為學(xué)特征影響的實(shí)驗(yàn)研究()
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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第29卷
期數(shù):
2017年07期
頁(yè)碼:
12-20,26
欄目:
基礎(chǔ)研究
出版日期:
2017-07-20

文章信息/Info

Title:
Effect of Bushen Zhuangjin Tang(補(bǔ)腎壯筋湯)on intervertebral disc and behavioral characteristics of rats with lumbar intervertebral disc degeneration
作者:
趙赫1; 高譽(yù)珊2; 胡振國(guó)1; 陳思學(xué)1; 劉濤1; 楊永棟1; 俞興1
1.北京中醫(yī)藥大學(xué)東直門(mén)醫(yī)院,北京 100700; 2.北京中醫(yī)藥大學(xué),北京 100029
Author(s):
ZHAO He1; GAO Yushan2; HU Zhenguo1; CHEN Sixue1; LIU Tao1; YANG Yongdong1; YU Xing1
1.Dongzhimen hospital of Beijing University of Traditional Chinese Medicine,Beijing 101121,China 2.Beijing University of Traditional Chinese Medicine,Beijing 100029,China
關(guān)鍵詞:
椎間盤(pán)退行性變;  動(dòng)物行為學(xué);  補(bǔ)腎壯筋湯;  大鼠; Sprague-Dawley;  動(dòng)物實(shí)驗(yàn)
Keywords:
Key words intervertebral disc degeneration;  ethology;  Bushen Zhuangjin Tang;  rats; Sprague-Dawley;  animal experimentation
摘要:
目的:觀察補(bǔ)腎壯筋湯對(duì)腰椎間盤(pán)退變大鼠椎間盤(pán)及行為學(xué)特征的影響,并觀察其安全性。方法:選取40只清潔級(jí)8周齡雌性SD大鼠,隨機(jī)分為假手術(shù)組、模型組、補(bǔ)腎壯筋湯組、吲哚美辛組,每組10只。應(yīng)用椎間盤(pán)穿刺法對(duì)模型組、補(bǔ)腎壯筋湯組、吲哚美辛組大鼠L5～6椎間盤(pán)進(jìn)行椎間盤(pán)退變?cè)炷?假手術(shù)組充分暴露L5～6椎間盤(pán)前緣,不進(jìn)行穿刺干預(yù)。造模后24 h,補(bǔ)腎壯筋湯組大鼠以補(bǔ)腎壯筋湯(1.5 g·kg-1)灌胃,吲哚美辛組大鼠以吲哚美辛膠囊水溶液(7.5 mg·kg-1)灌胃,假手術(shù)組、模型組大鼠均以生理鹽水(10 mL·kg-1)灌胃,每天1次,連續(xù)給藥7周。分別于造模前及藥物干預(yù)開(kāi)始后1、3、5、7周時(shí)進(jìn)行機(jī)械痛閾實(shí)驗(yàn)和斜板實(shí)驗(yàn)。最后一次行為學(xué)特征測(cè)定結(jié)束后,從眼底靜脈叢取血測(cè)定血清丙氨酸氨基轉(zhuǎn)移酶(alanine aminotransferase,AAT)、天冬氨酸氨基轉(zhuǎn)移酶(aspartate aminotransferase,AST)、肌酐(creatinine,CRE)、尿素氮(urea nitrogen,UN)的水平; 取L5～6椎間盤(pán)、肝臟及腎臟,椎間盤(pán)組織分別進(jìn)行HE染色、改良番紅O-固綠染色后觀察組織形態(tài)變化,肝臟及腎臟組織進(jìn)行HE染色、Masson染色后觀察組織病理改變。結(jié)果:①機(jī)械痛閾實(shí)驗(yàn)痛閾值。造模前及藥物干預(yù)開(kāi)始后不同時(shí)點(diǎn)痛閾值的差異有統(tǒng)計(jì)學(xué)意義,即存在時(shí)間效應(yīng)(F=36.245,P=0.000)。假手術(shù)組痛閾值隨時(shí)間變化趨勢(shì)不明顯; 模型組痛閾值隨時(shí)間變化逐漸降低; 補(bǔ)腎壯筋湯組和吲哚美辛組的痛閾值于藥物干預(yù)開(kāi)始后1 周時(shí)降至最低,后期變化不明顯。4組大鼠痛閾值總體比較,差異有統(tǒng)計(jì)學(xué)意義,即存在分組效應(yīng)(F=54.170,P=0.000)。除造模前外(F=1.375,P=0.266),藥物干預(yù)開(kāi)始后各時(shí)點(diǎn)4組大鼠痛閾值比較,組間差異均有統(tǒng)計(jì)學(xué)意義(F=30.059,P=0.000; F=34.117,P=0.000; F=33.454,P=0.000; F=35.617,P=0.000); 藥物干預(yù)開(kāi)始后各時(shí)點(diǎn)假手術(shù)組痛閾值均高于模型組(P=0.000; P=0.000; P=0.000; P=0.000); 除藥物干預(yù)開(kāi)始后1周時(shí)外(P=0.197,P=0.191),其余各時(shí)點(diǎn)模型組的痛閾值均低于補(bǔ)腎壯筋湯組和吲哚美辛組(P=0.010,P=0.013; P=0.000,P=0.000; P=0.000,P=0.000); 藥物干預(yù)開(kāi)始后各時(shí)點(diǎn)補(bǔ)腎壯筋湯組和吲哚美辛組痛閾值比較,組間差異均無(wú)統(tǒng)計(jì)學(xué)意義(P=0.100; P=0.067; P=0.108; P=0.097)。時(shí)間因素和分組因素存在交互效應(yīng)(F=16.490,P=0.000)。②斜板實(shí)驗(yàn)角度。造模前及藥物干預(yù)開(kāi)始后不同時(shí)點(diǎn)斜板實(shí)驗(yàn)角度的差異有統(tǒng)計(jì)學(xué)意義,即存在時(shí)間效應(yīng)(F=3.746,P=0.019)。假手術(shù)組、補(bǔ)腎壯筋湯組和吲哚美辛組斜板實(shí)驗(yàn)角度隨時(shí)間變化整體呈上升趨勢(shì); 模型組斜板實(shí)驗(yàn)角度隨時(shí)間變化整體呈下降趨勢(shì)。4組大鼠斜板實(shí)驗(yàn)角度總體比較,差異有統(tǒng)計(jì)學(xué)意義,即存在分組效應(yīng)(F=7.640,P=0.000)。造模前及藥物干預(yù)開(kāi)始后1周和3周時(shí),4組大鼠斜板實(shí)驗(yàn)角度比較,組間差異均無(wú)統(tǒng)計(jì)學(xué)意義(F=0.575,P=0.635; F=2.664,P=0.063; F=0.994,P=0.407)。藥物干預(yù)開(kāi)始后5周和7周時(shí),4組大鼠斜板實(shí)驗(yàn)角度比較,組間差異均有統(tǒng)計(jì)學(xué)意義(F=9.666,P=0.000; F=2.685,P=0.006); 假手術(shù)組斜板實(shí)驗(yàn)角度均大于模型組(P=0.000; P=0.026),模型組斜板實(shí)驗(yàn)角度均小于補(bǔ)腎壯筋湯組和吲哚美辛組(P=0.000,P=0.007; P=0.000,P=0.042),補(bǔ)腎壯筋湯組斜板實(shí)驗(yàn)角度均大于吲哚美辛組(P=0.000; P=0.047)。時(shí)間因素和分組因素存在交互效應(yīng)(F=2.934,P=0.012)。③血清學(xué)檢測(cè)結(jié)果。藥物干預(yù)結(jié)束后4組大鼠血清中AAT、AST、CRE、UN含量比較,組間差異均無(wú)統(tǒng)計(jì)學(xué)意義(F=1.023,P=0.398; F=0.237,P=0.869; F=0.629,P=0.602; F=1.141,P=0.350)。④組織學(xué)檢測(cè)結(jié)果。HE染色及改良番紅O-固綠染色顯示,與模型組相比,補(bǔ)腎壯筋湯組和吲哚美辛組大鼠腰椎間盤(pán)組織退變程度較輕,3組中補(bǔ)腎壯筋湯組椎間盤(pán)組織退變程度最輕; 肝、腎組織HE染色和Masson染色顯示,模型組、補(bǔ)腎壯筋湯組及吲哚美辛組均未見(jiàn)肝、腎組織纖維化。結(jié)論:補(bǔ)腎壯筋湯在延緩腰椎間盤(pán)退變大鼠椎間盤(pán)退變及改善其行為學(xué)特征方面具有一定作用,效果略優(yōu)于吲哚美辛,短期應(yīng)用不會(huì)對(duì)肝腎組織及肝腎功能造成損傷。
Abstract:
ABSTRACT Objective:To observe the effect of Bushen Zhuangjin Tang(補(bǔ)腎壯筋湯,BSZJT)on intervertebral disc and behavioral characteristics of rats with lumbar intervertebral disc degeneration and its safety.Methods:Forty 8-week-old clean-grade female SD rats were selected and randomly divided into sham-operated group,model group,BSZJT group and indometacin group,10 cases in each group.The models of L5/L6 intervertebral disc degeneration(IVDD)were built in rats in model group,BSZJT group and indometacin group by using intervertebral disc puncture method,while the rats in sham-operated group were treated with sham-operation to adequately expose the anterior border of L5/L6 intervertebral disc without any puncture intervention.At 24 hours after the modeling,the rats in BSZJT group and indometacin group were intragastric administrated with BSZJT(1.5 g/kg)and indometacin solution(7.5 mg/kg)respectively,while the rats in sham-operated group and model group were intragastric administrated with normal saline(10 mL·kg-1),once a day for consecutive 7 weeks.The mechanical paw withdrawal threshold(MPWT)test and the tiltboard test were carried out before modeling and at 1,3,5 and 7 weeks after the beginning of drug intervention respectively.The blood were fetched out from fundus oculi venous plexus after the last behavioral characteristics measurement,and the serum levels of alanine aminotransferase(AAT),aspartate aminotransferase(AST),creatinine(CRE)and urea nitrogen(UN)were measured.The L5/L6 intervertebral discs,livers and kidneys were fetched out.The intervertebral disc tissues were received HE staining and improved safranin O-fast green staining respectively and the hepatic tissues and nephridial tissues were received HE staining and Masson staining for observing the histopathological changes.Results:There was statistical difference in the pain threshold values between different timepoints before modeling and after the beginning of drug intervention,in other words,there was time effect(F=36.245,P=0.000).The pain threshold value changed unconspicuously with time in sham-operated group and the pain threshold value decreased gradually with time in model group.The lowest pain threshold value was found at 1 week after the beginning of drug intervention and the pain threshold value changed unconspicuously with time in later period in BSZJT group and indometacin group.There was statistical difference in the pain threshold values between the 4 groups in general,in other words,there was group effect(F=54.170,P=0.000).There was not statistical difference in the pain threshold values between the 4 groups before the modeling(F=1.375,P=0.266),and there was statistical difference in the pain threshold values between the 4 groups at each timepoint after the beginning of drug intervention(F=30.059,P=0.000; F=34.117,P=0.000; F=33.454,P=0.000; F=35.617,P=0.000).The pain threshold values were higher in sham-operated group compared to model group at each timepoint after the beginning of drug intervention(P=0.000; P=0.000; P=0.000; P=0.000).The pain threshold values were lower in model group compared to BSZJT group and indometacin group(P=0.010,P=0.013; P=0.000,P=0.000; P=0.000,P=0.000)except at 1 week after the beginning of drug intervention(P=0.197,P=0.191).There was no statistical difference in the pain threshold values between BSZJT group and indometacin group at each timepoint after the beginning of drug intervention(P=0.100; P=0.067; P=0.108; P=0.097).There was interaction between time factor and grouping factor(F=16.490,P=0.000).There was statistical difference in the tiltboard test angle between different timepoints before modeling and after the beginning of drug intervention,in other words,there was time effect(F=3.746,P=0.019).The tiltboard test angle increased with time in general in sham-operated group,BSZJT group and indometacin group,while the tiltboard test angle decreased with time in general in model group.There was statistical difference in the tiltboard test angle between the 4 groups in general,in other words,there was group effect(F=7.640,P=0.000).There was no statistical difference in the tiltboard test angle between the 4 groups before modeling and at 1 and 3 weeks after the beginning of drug intervention(F=0.575,P=0.635; F=2.664,P=0.063; F=0.994,P=0.407).There was statistical difference in the tiltboard test angle between the 4 groups at 5 and 7 weeks after the beginning of drug intervention(F=9.666,P=0.000; F=2.685,P=0.006).The tiltboard test angle was larger in sham-operated group compared to model group(P=0.000; P=0.026)and was smaller in model group compared to BSZJT group and indometacin group(P=0.000,P=0.007; P=0.000,P=0.042)and was larger in BSZJT group compared to indometacin group(P=0.000; P=0.047).There was interaction between time factor and grouping factor(F=2.934,P=0.012).There was no statistical difference in the serum contents of AAT,AST,CRE and UN between the 4 groups after the end of drug intervention(F=1.023,P=0.398; F=0.237,P=0.869; F=0.629,P=0.602; F=1.141,P=0.35).HE staining results and improved safranin O-fast green staining results showed the degeneration degree of lumbar intervertebral disc tissue was slighter in BSZJT group and indometacin group compared to model group and the degeneration degree was slightest in BSZJT group.HE staining results and Masson staining results showed that no fibrosis were found in hepatic tissues and nephridial tissues in model group,BSZJT group and indometacin group.Conclusion:BSZJT can delay intervertebral disc degeneration and improve the behavioral characteristics to some extent in rats with lumbar intervetebral disc degeneration,and it slightly surpasses indometacin in the effect.Moreover,short-term application of BSZJT can not cause damage to hepatorenal tissues and hepatorenal function.

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通訊作者:俞興 E-mail:[email protected]
更新日期/Last Update: 2017-12-29