[1]赵赫,高誉珊,胡振国,等.补肾壮筋汤对腰椎间盘退变大鼠椎间盘及行为学特征影响的实验研究[J].中医正骨,2017,29(07):12-20,26.
　ZHAO He,GAO Yushan,HU Zhenguo,et al.Effect of Bushen Zhuangjin Tang(补肾壮筋汤)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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补肾壮筋汤对腰椎间盘退变大鼠椎间盘及行为学特征影响的实验研究()
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《中医正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第29卷
期数:
2017年07期
页码:
12-20,26
栏目:
基础研究
出版日期:
2017-07-20

文章信息/Info

Title:
Effect of Bushen Zhuangjin Tang(补肾壮筋汤)on intervertebral disc and behavioral characteristics of rats with lumbar intervertebral disc degeneration
作者:
赵赫1; 高誉珊2; 胡振国1; 陈思学1; 刘涛1; 杨永栋1; 俞兴1
1.北京中医药大学东直门医院,北京 100700; 2.北京中医药大学,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
关键词:
椎间盘退行性变;  动物行为学;  补肾壮筋汤;  大鼠; Sprague-Dawley;  动物实验
Keywords:
Key words intervertebral disc degeneration;  ethology;  Bushen Zhuangjin Tang;  rats; Sprague-Dawley;  animal experimentation
摘要:
目的:观察补肾壮筋汤对腰椎间盘退变大鼠椎间盘及行为学特征的影响,并观察其安全性。方法:选取40只清洁级8周龄雌性SD大鼠,随机分为假手术组、模型组、补肾壮筋汤组、吲哚美辛组,每组10只。应用椎间盘穿刺法对模型组、补肾壮筋汤组、吲哚美辛组大鼠L5～6椎间盘进行椎间盘退变造模,假手术组充分暴露L5～6椎间盘前缘,不进行穿刺干预。造模后24 h,补肾壮筋汤组大鼠以补肾壮筋汤(1.5 g·kg-1)灌胃,吲哚美辛组大鼠以吲哚美辛胶囊水溶液(7.5 mg·kg-1)灌胃,假手术组、模型组大鼠均以生理盐水(10 mL·kg-1)灌胃,每天1次,连续给药7周。分别于造模前及药物干预开始后1、3、5、7周时进行机械痛阈实验和斜板实验。最后一次行为学特征测定结束后,从眼底静脉丛取血测定血清丙氨酸氨基转移酶(alanine aminotransferase,AAT)、天冬氨酸氨基转移酶(aspartate aminotransferase,AST)、肌酐(creatinine,CRE)、尿素氮(urea nitrogen,UN)的水平; 取L5～6椎间盘、肝脏及肾脏,椎间盘组织分别进行HE染色、改良番红O-固绿染色后观察组织形态变化,肝脏及肾脏组织进行HE染色、Masson染色后观察组织病理改变。结果:①机械痛阈实验痛阈值。造模前及药物干预开始后不同时点痛阈值的差异有统计学意义,即存在时间效应(F=36.245,P=0.000)。假手术组痛阈值随时间变化趋势不明显; 模型组痛阈值随时间变化逐渐降低; 补肾壮筋汤组和吲哚美辛组的痛阈值于药物干预开始后1 周时降至最低,后期变化不明显。4组大鼠痛阈值总体比较,差异有统计学意义,即存在分组效应(F=54.170,P=0.000)。除造模前外(F=1.375,P=0.266),药物干预开始后各时点4组大鼠痛阈值比较,组间差异均有统计学意义(F=30.059,P=0.000; F=34.117,P=0.000; F=33.454,P=0.000; F=35.617,P=0.000); 药物干预开始后各时点假手术组痛阈值均高于模型组(P=0.000; P=0.000; P=0.000; P=0.000); 除药物干预开始后1周时外(P=0.197,P=0.191),其余各时点模型组的痛阈值均低于补肾壮筋汤组和吲哚美辛组(P=0.010,P=0.013; P=0.000,P=0.000; P=0.000,P=0.000); 药物干预开始后各时点补肾壮筋汤组和吲哚美辛组痛阈值比较,组间差异均无统计学意义(P=0.100; P=0.067; P=0.108; P=0.097)。时间因素和分组因素存在交互效应(F=16.490,P=0.000)。②斜板实验角度。造模前及药物干预开始后不同时点斜板实验角度的差异有统计学意义,即存在时间效应(F=3.746,P=0.019)。假手术组、补肾壮筋汤组和吲哚美辛组斜板实验角度随时间变化整体呈上升趋势; 模型组斜板实验角度随时间变化整体呈下降趋势。4组大鼠斜板实验角度总体比较,差异有统计学意义,即存在分组效应(F=7.640,P=0.000)。造模前及药物干预开始后1周和3周时,4组大鼠斜板实验角度比较,组间差异均无统计学意义(F=0.575,P=0.635; F=2.664,P=0.063; F=0.994,P=0.407)。药物干预开始后5周和7周时,4组大鼠斜板实验角度比较,组间差异均有统计学意义(F=9.666,P=0.000; F=2.685,P=0.006); 假手术组斜板实验角度均大于模型组(P=0.000; P=0.026),模型组斜板实验角度均小于补肾壮筋汤组和吲哚美辛组(P=0.000,P=0.007; P=0.000,P=0.042),补肾壮筋汤组斜板实验角度均大于吲哚美辛组(P=0.000; P=0.047)。时间因素和分组因素存在交互效应(F=2.934,P=0.012)。③血清学检测结果。药物干预结束后4组大鼠血清中AAT、AST、CRE、UN含量比较,组间差异均无统计学意义(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组织学检测结果。HE染色及改良番红O-固绿染色显示,与模型组相比,补肾壮筋汤组和吲哚美辛组大鼠腰椎间盘组织退变程度较轻,3组中补肾壮筋汤组椎间盘组织退变程度最轻; 肝、肾组织HE染色和Masson染色显示,模型组、补肾壮筋汤组及吲哚美辛组均未见肝、肾组织纤维化。结论:补肾壮筋汤在延缓腰椎间盘退变大鼠椎间盘退变及改善其行为学特征方面具有一定作用,效果略优于吲哚美辛,短期应用不会对肝肾组织及肝肾功能造成损伤。
Abstract:
ABSTRACT Objective:To observe the effect of Bushen Zhuangjin Tang(补肾壮筋汤,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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通讯作者:俞兴 E-mail:yuxing34@sina.com
更新日期/Last Update: 2017-12-29