中药热敷颈椎病模型兔椎间盘自噬及凋亡相关因子的表达

Effect of traditional Chinese medicine hot compress on autophagy and expression of apoptosis related factors in the intervertebral disc of rabbits with cervical spondylosis

背景:中药热敷作为治疗颈椎病的常用方法,具有操作便捷、临床疗效稳定、不良作用少的优势,但其效应机制尚不明确。目的:观察中药热敷对颈椎病模型兔椎间盘自噬及凋亡相关因子的影响,探讨其治疗颈椎病的作用机制。方法:将24只新西兰兔随机分为对照组、模型组、中药热敷组,每组各8只。除对照组外,其余两组建立颈椎病兔模型。通过对比造模前、后颈椎影像学变化,观察是否有颈椎生理曲度变化、椎间孔变小、椎间隙变窄以及骨赘生成等改变,作为造模评价标准。动物模型建成后,中药热敷组予中药热敷干预治疗,20 min/次,每5 min更换一次药包,每日治疗一次,疗程共20次;其余两组正常饲养,不做干预。通过苏木精-伊红染色法观察各组兔颈椎间盘组织形态改变;采用TUNEL法检测颈椎间盘细胞凋亡情况;运用qRT-PCR法测定颈椎间盘组织Bcl-2、Beclin1、微管相关蛋白1轻链3 mRNA的相对表达水平。结果与结论:①模型组兔颈椎间盘组织形态学评分比对照组高(P<0.05),中药热敷组椎间盘组织形态学评分比模型组低(P<0.05);②模型组兔颈椎间盘中细胞凋亡率较对照组增高(P<0.05),中药热敷组兔颈椎间盘中细胞凋亡率较模型组下降(P<0.05);③模型组兔颈椎间盘自噬及凋亡相关因子Bcl-2、Beclin1、微管相关蛋白1轻链3 mRNA的相对表达水平较对照组降低(P<0.05),而中药热敷组兔颈椎间盘中Bcl-2、Beclin1、微管相关蛋白1轻链3 mRNA的相对表达水平高于模型组(P<0.05);④提示中药热敷可以增加颈椎病模型兔椎间盘自噬因子的表达,减少椎间盘细胞凋亡,从而保护椎间盘,改善颈椎病。

BACKGROUND: Traditional Chinese medicine hot compress is a common method to treat cervical spondylosis. It has the unique advantages of convenient operation, stable curative effect and few side effects, but its effect mechanism is not clear.OBJECTIVE: To observe the effect of traditional Chinese medicine hot compress on autophagy and expression of apoptosis related factors in the intervertebral disc of rabbits with cervical spondylosis and to explore its mechanism in the treatment of cervical spondylosis.METHODS: Twenty-four New Zealand white rabbits were randomly divided into control group, model group, and traditional Chinese medicine hot compress group(drug group), with eight rabbits in each group. Except for the control group, 16 rabbits in the other two groups were used to establish the animal model of cervical spondylosis. The imaging changes of the cervical spine before and a?ter modeling were compared to observe whether there were changes in the physiological curvature of the cervical spine, smaller intervertebral foramen, narrowing of the intervertebral space, and the formation of osteophytes, which could be used as the evaluation standard of modeling. A?ter the animal model was established, the drug group was treated with traditional Chinese medicine hot compress intervention, 20 minutes per day, and the medicine bag was changed every 5 minutes, once a day, for 20 days in total. The other two groups were fed normally without intervention. The morphological changes of the cervical intervertebral disc of rabbits in each group were observed by hematoxylineosin staining. The apoptosis of intervertebral disc cells in each group was observed by TUNEL. The relative mRNA expression levels of Bcl-2, Beclin1, and microtubule-associated protein 1 light chain 3 in the cervical intervertebral disc were measured by real-time fluorescence quantitative PCR.RESULTS AND CONCLUSION: The histological score of the cervical intervertebral disc in the model group was higher than that in the control group(P < 0.05),and the histological score of the intervertebral disc in the traditional Chinese medicine hot compress group was lower than that in the model group(P < 0.05).The apoptosis rate of rabbit cervical intervertebral disc in the model group was higher than that in the control group(P < 0.05), and the apoptosis rate of rabbit cervical intervertebral disc in the traditional Chinese medicine hot compress group was lower than that in the model group(P < 0.05). The relative mRNA expression levels of Bcl-2, Beclin1, and microtubule-associated protein 1 light chain 3 in the model group were lower than those in the control group(P < 0.05),while the relative mRNA expression levels of Bcl-2, Beclin1, and microtubule-associated protein 1 light chain 3 in the traditional Chinese medicine hot compress group were significantly higher than those in the model group(P < 0.05). To conclude, traditional Chinese medicine hot compress can increase the expression of autophagy factors in the intervertebral disc of rabbits with cervical spondylosis and reduce the apoptosis of intervertebral disc cells, so as to protect the intervertebral disc and improve cervical spondylosis.operation,stable curative effect and few side effects,but its effect mechanism is not clear.OBJECTIVE:To observe the effect of traditional Chinese medicine hot compress on autophagy and expression of apoptosis related factors in the intervertebral disc of rabbits with ce rvical spondylosis and to explore its mechanism in the treatment of cervical spondylosis.METHODS:Twenty-four New Zealand white rabbits were randomly divided into control group,model group,and traditional Chinese medicine hot compress group(drug group),with eight rabbits in each group.Except for the control group,16 rabbits in the other two groups were used to establish the animal model of cervical spondylosis.The imaging changes of the ce rvical spine before and after modeling were compared to observe whether there were changes in the physiological curvature of the cervical spine,smaller interve rtebral fo ramen,narrowing of the intervertebral space,and the formation of osteophytes,which co uld be used as the evaluation standard of modeling.After the animal model was established,the drug group was treated with traditional Chinese medicine hot compress intervention,20 minutes per day,and the medicine bag was changed every 5 minutes,once a day,for 20 days in total.The other two groups were fed normally without intervention.The morphological changes of the cervical interve rtebral disc of rabbits in each group were observed by hematoxylineosin staining.The apoptosis of inte rvertebral disc cells in each group was observed by TUNEL.The relative mRNA expression levels of Bcl-2,Beclinl,and microtubule-associated protein 1 light chain 3 in the cervical intervertebral disc were measured by real-time fluorescence quantitative PCR.RESULTS AND CONCLUSION:The histological score of the cervical inte rvertebral disc in the model group was higher than that in the control group(P <0.05),and the histological score of the intervertebral disc in the traditional Chinese medicine hot compress group was lower than that in the model group(P <0.05).The apoptosis rate of rabbit ce rvical interve rtebral disc in the model group was higher than that in the control group(P <0.05),and the apoptosis rate of rabbit cervical intervertebral disc in the tra ditional Chinese medicine hot compress group was lower than that in the model group(P <0.05).The relative mRNA expression levels of Bcl-2,Beclin1,and microtubule-associated protein 1 light chain 3 in the model group were lower than those in the control group(P <0.05),while the relative mRNA expression levels of Bcl-2,Beclin1,and mic rotubule-associated protein 1 light chain 3 in the traditional Chinese medicine hot compress group were significantly higher than those in the model group(P <0.05).To conclude,traditional Chinese medicine hot compress can increase the expression of autophagy factors in the intervertebral disc of rabbits with cervical spondylosis and reduce the apoptosis of intervertebral disc cells,so as to protect the interve rtebral disc and improve cervical spondylosis.