点拨腰大肌对纤维环损伤大鼠模型腰大肌被动拉伸张力影响的研究
发布时间：2018-09-11 08:36
【摘要】：腰痛是临床常见病、多发病、疑难病,已严重影响患者的工作与生活,近年来椎间盘源性腰痛的发病率日益增高,成为目前临床腰痛研究领域的热点与难点。椎间盘源性腰痛(Lumbar discogenic pain, LDP)指的是起源于椎间盘本身的疼痛,以腰腹痛、腰臀痛、腰痛连及腹股沟部广泛性疼痛为典型症状,这与中医所描述的“带脉瘀滞型腰痛”的典型表现——“腹腰脊痛,冲阴股”很相仿。触诊时发现患者除了棘突间压痛以外,还存在髂腰肌,尤其是腰大肌的痉挛疼痛、肌张力增高,也就是说这类腰痛除有椎间盘本身的病变外,可能还伴有脊旁肌的病理改变。在临床诊疗时发现,点拨腰大肌干预可有效改善临床症状。结合脊柱静动力平衡理论,我们认为LDP的发生可能是由脊柱静力结构失衡与动力结构失衡相互影响造成的,而对动力结构脊旁肌的干预可能会促进静力结构椎间盘损伤的修复。 研究目的 本实验拟通过针刺纤维环建立腰椎间盘损伤模型,观察该模型是否伴有腰大肌的形态学及被动拉伸特性的变化以及点拨腰大肌对这些变化的影响。阐释点拨腰大肌治疗椎间盘源性腰痛的机制,为探寻推拿带脉治疗腰痛的新方法提供实验依据。 研究方法 实验一： Wistar雄性7周龄大鼠30只,按随机数字表法分为造模组20只,假手术组10只。造模组大鼠行L4-5、L5-6椎间盘前入路纤维环全层针刺手术,制作纤维环针刺损伤模型；假手术组不行针刺。造模手术后,再将造模组按随机数字表法平均分为模型组和治疗组。 造模手术后2周,将治疗组大鼠装入自制鼠袋,取俯卧位,做手法点拨双侧腰大肌治疗干预。隔日治疗1次,共治疗10次(约3周)。模型组大鼠和假手术组大鼠只装袋,不做治疗。 利用动物自主行为研究仪观察大鼠造模前、造模后及治疗后的行为学变化。在造模前随机选取10只大鼠做垂直活动度观察；造模手术后2周,对假手术组、造模组随机选取10只大鼠再次进行垂直活动度观察1次；治疗10次之后,再次对假手术组、模型组和治疗组进行垂直活动度观察1次,以观察大鼠行为学改变。 治疗结束后,所有大鼠麻醉处死,取双侧腰大肌行被动拉伸测试；然后取L4-5、L5-6节段椎间盘,包埋,切片,行HE染色,光镜下观察其组织形态学改变；以免疫组织化学法,探测椎间盘内炎症因子IL-1p的阳性表达。 实验二： Wistar雄性大鼠8只,随机分为造模组和假手术组,其中造模组6只,假手术组2只。同法制作模型。造模手术后,再将造模组按随机数字表法分为1周模型组、3周模型组、6周模型组,各2只。并分别于手术后1周、3周、6周取双侧腰大肌。 对腰大肌行HE染色,观察肌肉形态学变化；行苦味酸天狼星红染色,观察腰大肌内胶原性质及含量。 研究结果 1垂直活动度检测： 造模后5周：模型组与假手术组相比,垂直站立红外线计数减少,且差异有统计学意义(P0.05)；治疗组与模型组相比,垂直站立红外线计数增多,但差异无统计学意义(P0.05)。提示椎间盘损伤5周后大鼠垂直站立活动度降低。 2椎间盘HE染色： 造模后5周：髓核：假手术组呈中央密集型分布,在四周呈现出均匀分布状态,髓核细胞数目较多；模型组髓核组织稀疏,分布不均,伴有大量不定形物质；治疗组与假手术组相近。纤维环：假手术组可见纤维环层次分明,排列整齐,与髓核界线明显；模型组纤维环可见纤维环排列轻度改变,有紊乱倾向；治疗组纤维环形态接近假手术组。提示针刺损伤纤维环能够造成椎间盘一定程度的损伤,点拨腰大肌治疗对于椎间盘纤维环损伤可能有良性干预作用。 3椎间盘IL-1β免疫组织化学： 在纤维环细胞可检测到椎间盘炎症因子IL-1β的阳性表达,阳性细胞呈棕黄色。 三组间免疫组化阳性表达积分光密度(IOD)差异有统计学意义(P0.05)；相对于假手术组,模型组阳性表达IOD值明显增强(P0.01)；治疗组与模型组相比,阳性表达IOD值减少明显,差异具有统计学意义(P0.01)。提示针刺损伤纤维环后,会造成炎性反应,且点拨腰大肌治疗对椎间盘炎性改变具有良性干预作用。 4腰大肌拉伸力学： 左侧腰大肌弹性模量对比：模型组弹性模量较假手术组降低,治疗组较模型组增高,三组间差异无统计学意义(P0.05)。右侧腰大肌弹性模量对比：三组间差异具有统计学意义(P0.05)；模型组较假手术组明显减少,差异具有统计学意义(P0.01)；治疗组较模型组有所增加,但差异无统计学意义(P0.05)。提示针刺损伤椎间盘后,大鼠右侧腰大肌弹性模量出现一定程度的降低,点拨腰大肌治疗对腰大肌弹性模量可能具有一定的干预作用。 5腰大肌HE染色： 1周腰大肌可见肌纤维有序排列,肌束紧密,其中细胞核呈椭圆形,大部分细胞核沿肌纤维的长轴边缘排列,少数细胞核位于肌纤维内；3周腰大肌肌纤维开始出现退变,肌纤维轻度肿胀,肌横纹模糊,肌浆呈绒毛状或融合性改变；6周肌纤维退变加重,肌横纹模糊或消失,嗜酸性增强,肌浆绒毛状或融合性改变明显,肌纤维散乱无序甚至出现节段样变。 6天狼星红苦味酸染色：模型组腰大肌内胶原含量较假手术组发生一定程度改变。 研究结论 1垂直站立红外线计数、椎间盘形态学、椎间盘炎症因子IL-1β及腰大肌的拉伸力学、形态学等改变能够说明,椎间盘纤维环针刺损伤大鼠模型的腰大肌弹性模量会发生一定的改变。 2干预治疗后,针刺损伤大鼠模型的腰大肌弹性模量有所恢复。推拿手法对椎间盘纤维环、髓核组织形态及炎症因子IL-1p表达可能具有良性调节作用。
[Abstract]:Lumbar discogenic pain (LDP) refers to the pain originating from the intervertebral disc itself to the lumbar abdomen. Pain, lumbogluteal pain, low back pain and generalized pain in the groin are typical symptoms, which are similar to the typical manifestation of "lumbago with blood stasis" described by traditional Chinese medicine - "abdominal low back pain, Chongyin femur". That is to say, in addition to the pathological changes of the intervertebral disc itself, this kind of low back pain may also be accompanied by pathological changes of the paraspinal muscles. The intervention of dynamic structure of the paraspinal muscles may promote the repair of static structural intervertebral disc injury.
research objective
The purpose of this study is to establish a model of lumbar intervertebral disc injury by needling fibrous rings to observe whether the model is accompanied by changes in morphology and passive stretching properties of psoas major muscle and the effect of tapping psoas major muscle on these changes. Experimental basis.
research method
Experiment 1:
30 Wistar male rats aged 7 weeks were divided into model group (20 rats) and sham operation group (10 rats) according to random number table method. And treatment group.
Two weeks after the operation, the rats in the treatment group were placed in a self-made pouch and placed in a prone position. The rats in the model group and the sham operation group were treated once every other day for 10 times (about 3 weeks).
The behavioral changes of rats before, after and after modeling were observed by animal autonomous behavior research instrument.Ten rats were randomly selected to observe the vertical mobility before modeling. The rats in the operation group, the model group and the treatment group were observed 1 times of vertical activity to observe the behavioral changes in rats.
After the treatment, all rats were anesthetized and executed, bilateral psoas major muscles were taken for passive stretching test, L4-5 and L5-6 intervertebral discs were embedded, sliced, stained with HE, and the histomorphological changes were observed under light microscope. The positive expression of inflammatory factor IL-1p in intervertebral discs was detected by immunohistochemistry.
Experiment two:
Eight male Wistar rats were randomly divided into model group and sham-operation group, of which 6 were in the model group and 2 were in the sham-operation group.
HE staining was used to observe the morphological changes of psoas major muscle and Sirius picrate red staining was used to observe the properties and content of collagen in psoas major muscle.
Research results
1 vertical activity measurement:
Five weeks after modeling: Compared with the sham operation group, the vertical standing infrared counts of the model group decreased, and the difference was statistically significant (P 0.05); Compared with the model group, the vertical standing infrared counts of the treatment group increased, but the difference was not statistically significant (P 0.05). It suggested that the vertical standing mobility of the rats decreased after 5 weeks of intervertebral disc injury.
2 intervertebral disc HE staining:
5 weeks after modeling: nucleus pulposus: sham-operated group showed a centrally dense distribution around the state of uniform distribution, the number of nucleus pulposus cells more; model group nucleus pulposus tissue sparse, uneven distribution, with a large number of amorphous substances; treatment group and sham-operated group similar. The annulus fibrosus in the treatment group was similar to that in the sham-operated group, suggesting that needling injury of the annulus fibrosus could cause a certain degree of injury of the intervertebral disc, and the treatment of poking psoas major muscle may have a benign intervention effect on the annulus fibrosus injury of the intervertebral disc.
3 IL-1 immunohistochemistry of intervertebral disc:
Positive expression of inflammatory factor IL-1 beta was detected in annulus fibrosus cells, and the positive cells were brown.
Compared with sham operation group, the positive expression of IOD in model group increased significantly (P 0.01). Compared with model group, the positive expression of IOD in treatment group decreased significantly (P 0.01). The difference was statistically significant (P 0.01). The treatment of lumbar disc muscle is effective in the treatment of inflammatory changes of the intervertebral disc.
4 the stretching mechanics of psoas major muscle:
Comparing the elastic modulus of left psoas major muscle: The elastic modulus of model group was lower than that of sham operation group, and that of treatment group was higher than that of model group. There was no significant difference among three groups (P 0.05). 1) Compared with the model group, the treatment group increased, but there was no significant difference (P 0.05).
5 HE staining of psoas major muscle:
At 1 week, the psoas major muscle fibers were arranged orderly and the muscle bundles were compact. The nuclei were elliptical, most of the nuclei were arranged along the long axis edge of the muscle fibers, and a few nuclei were located in the muscle fibers. Vitamin degeneration aggravated, myofibrils blurred or disappeared, eosinophilic increased, myofibrillar or fusion changes were obvious, myofibrils disordered and even segmental changes.
Sirius red picric acid staining: The collagen content in psoas major muscle of model group changed to a certain extent than that of sham operation group.
research conclusion
1 Vertical standing infrared counting, intervertebral disc morphology, intervertebral disc inflammatory factor IL-1beta and the tensile mechanics and morphology of psoas major muscle can explain that the elastic modulus of psoas major muscle in the rat model of intervertebral disc annulus acupuncture injury will change.
After intervention treatment, the elastic modulus of psoas major muscle of the rats with acupuncture injury recovered. Massage manipulation may have a benign regulatory effect on the fibrous ring of intervertebral disc, the morphology of nucleus pulposus and the expression of inflammatory factor IL-1p.
【学位授予单位】：北京中医药大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R245
本文编号：2236172
[Abstract]:Lumbar discogenic pain (LDP) refers to the pain originating from the intervertebral disc itself to the lumbar abdomen. Pain, lumbogluteal pain, low back pain and generalized pain in the groin are typical symptoms, which are similar to the typical manifestation of "lumbago with blood stasis" described by traditional Chinese medicine - "abdominal low back pain, Chongyin femur". That is to say, in addition to the pathological changes of the intervertebral disc itself, this kind of low back pain may also be accompanied by pathological changes of the paraspinal muscles. The intervention of dynamic structure of the paraspinal muscles may promote the repair of static structural intervertebral disc injury.
research objective
The purpose of this study is to establish a model of lumbar intervertebral disc injury by needling fibrous rings to observe whether the model is accompanied by changes in morphology and passive stretching properties of psoas major muscle and the effect of tapping psoas major muscle on these changes. Experimental basis.
research method
Experiment 1:
30 Wistar male rats aged 7 weeks were divided into model group (20 rats) and sham operation group (10 rats) according to random number table method. And treatment group.
Two weeks after the operation, the rats in the treatment group were placed in a self-made pouch and placed in a prone position. The rats in the model group and the sham operation group were treated once every other day for 10 times (about 3 weeks).
The behavioral changes of rats before, after and after modeling were observed by animal autonomous behavior research instrument.Ten rats were randomly selected to observe the vertical mobility before modeling. The rats in the operation group, the model group and the treatment group were observed 1 times of vertical activity to observe the behavioral changes in rats.
After the treatment, all rats were anesthetized and executed, bilateral psoas major muscles were taken for passive stretching test, L4-5 and L5-6 intervertebral discs were embedded, sliced, stained with HE, and the histomorphological changes were observed under light microscope. The positive expression of inflammatory factor IL-1p in intervertebral discs was detected by immunohistochemistry.
Experiment two:
Eight male Wistar rats were randomly divided into model group and sham-operation group, of which 6 were in the model group and 2 were in the sham-operation group.
HE staining was used to observe the morphological changes of psoas major muscle and Sirius picrate red staining was used to observe the properties and content of collagen in psoas major muscle.
Research results
1 vertical activity measurement:
Five weeks after modeling: Compared with the sham operation group, the vertical standing infrared counts of the model group decreased, and the difference was statistically significant (P 0.05); Compared with the model group, the vertical standing infrared counts of the treatment group increased, but the difference was not statistically significant (P 0.05). It suggested that the vertical standing mobility of the rats decreased after 5 weeks of intervertebral disc injury.
2 intervertebral disc HE staining:
5 weeks after modeling: nucleus pulposus: sham-operated group showed a centrally dense distribution around the state of uniform distribution, the number of nucleus pulposus cells more; model group nucleus pulposus tissue sparse, uneven distribution, with a large number of amorphous substances; treatment group and sham-operated group similar. The annulus fibrosus in the treatment group was similar to that in the sham-operated group, suggesting that needling injury of the annulus fibrosus could cause a certain degree of injury of the intervertebral disc, and the treatment of poking psoas major muscle may have a benign intervention effect on the annulus fibrosus injury of the intervertebral disc.
3 IL-1 immunohistochemistry of intervertebral disc:
Positive expression of inflammatory factor IL-1 beta was detected in annulus fibrosus cells, and the positive cells were brown.
Compared with sham operation group, the positive expression of IOD in model group increased significantly (P 0.01). Compared with model group, the positive expression of IOD in treatment group decreased significantly (P 0.01). The difference was statistically significant (P 0.01). The treatment of lumbar disc muscle is effective in the treatment of inflammatory changes of the intervertebral disc.
4 the stretching mechanics of psoas major muscle:
Comparing the elastic modulus of left psoas major muscle: The elastic modulus of model group was lower than that of sham operation group, and that of treatment group was higher than that of model group. There was no significant difference among three groups (P 0.05). 1) Compared with the model group, the treatment group increased, but there was no significant difference (P 0.05).
5 HE staining of psoas major muscle:
At 1 week, the psoas major muscle fibers were arranged orderly and the muscle bundles were compact. The nuclei were elliptical, most of the nuclei were arranged along the long axis edge of the muscle fibers, and a few nuclei were located in the muscle fibers. Vitamin degeneration aggravated, myofibrils blurred or disappeared, eosinophilic increased, myofibrillar or fusion changes were obvious, myofibrils disordered and even segmental changes.
Sirius red picric acid staining: The collagen content in psoas major muscle of model group changed to a certain extent than that of sham operation group.
research conclusion
1 Vertical standing infrared counting, intervertebral disc morphology, intervertebral disc inflammatory factor IL-1beta and the tensile mechanics and morphology of psoas major muscle can explain that the elastic modulus of psoas major muscle in the rat model of intervertebral disc annulus acupuncture injury will change.
After intervention treatment, the elastic modulus of psoas major muscle of the rats with acupuncture injury recovered. Massage manipulation may have a benign regulatory effect on the fibrous ring of intervertebral disc, the morphology of nucleus pulposus and the expression of inflammatory factor IL-1p.
【学位授予单位】：北京中医药大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R245
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