點撥腰大肌對纖維環(huán)損傷大鼠模型腰大肌被動拉伸張力影響的研究
發(fā)布時間：2018-09-11 08:36
【摘要】：腰痛是臨床常見病、多發(fā)病、疑難病,已嚴重影響患者的工作與生活,近年來椎間盤源性腰痛的發(fā)病率日益增高,成為目前臨床腰痛研究領(lǐng)域的熱點與難點。椎間盤源性腰痛(Lumbar discogenic pain, LDP)指的是起源于椎間盤本身的疼痛,以腰腹痛、腰臀痛、腰痛連及腹股溝部廣泛性疼痛為典型癥狀,這與中醫(yī)所描述的“帶脈瘀滯型腰痛”的典型表現(xiàn)——“腹腰脊痛,沖陰股”很相仿。觸診時發(fā)現(xiàn)患者除了棘突間壓痛以外,還存在髂腰肌,尤其是腰大肌的痙攣疼痛、肌張力增高,也就是說這類腰痛除有椎間盤本身的病變外,可能還伴有脊旁肌的病理改變。在臨床診療時發(fā)現(xiàn),點撥腰大肌干預(yù)可有效改善臨床癥狀。結(jié)合脊柱靜動力平衡理論,我們認為LDP的發(fā)生可能是由脊柱靜力結(jié)構(gòu)失衡與動力結(jié)構(gòu)失衡相互影響造成的,而對動力結(jié)構(gòu)脊旁肌的干預(yù)可能會促進靜力結(jié)構(gòu)椎間盤損傷的修復(fù)。 研究目的 本實驗擬通過針刺纖維環(huán)建立腰椎間盤損傷模型,觀察該模型是否伴有腰大肌的形態(tài)學及被動拉伸特性的變化以及點撥腰大肌對這些變化的影響。闡釋點撥腰大肌治療椎間盤源性腰痛的機制,為探尋推拿帶脈治療腰痛的新方法提供實驗依據(jù)。 研究方法 實驗一： Wistar雄性7周齡大鼠30只,按隨機數(shù)字表法分為造模組20只,假手術(shù)組10只。造模組大鼠行L4-5、L5-6椎間盤前入路纖維環(huán)全層針刺手術(shù),制作纖維環(huán)針刺損傷模型；假手術(shù)組不行針刺。造模手術(shù)后,再將造模組按隨機數(shù)字表法平均分為模型組和治療組。 造模手術(shù)后2周,將治療組大鼠裝入自制鼠袋,取俯臥位,做手法點撥雙側(cè)腰大肌治療干預(yù)。隔日治療1次,共治療10次(約3周)。模型組大鼠和假手術(shù)組大鼠只裝袋,不做治療。 利用動物自主行為研究儀觀察大鼠造模前、造模后及治療后的行為學變化。在造模前隨機選取10只大鼠做垂直活動度觀察；造模手術(shù)后2周,對假手術(shù)組、造模組隨機選取10只大鼠再次進行垂直活動度觀察1次；治療10次之后,再次對假手術(shù)組、模型組和治療組進行垂直活動度觀察1次,以觀察大鼠行為學改變。 治療結(jié)束后,所有大鼠麻醉處死,取雙側(cè)腰大肌行被動拉伸測試；然后取L4-5、L5-6節(jié)段椎間盤,包埋,切片,行HE染色,光鏡下觀察其組織形態(tài)學改變；以免疫組織化學法,探測椎間盤內(nèi)炎癥因子IL-1p的陽性表達。 實驗二： Wistar雄性大鼠8只,隨機分為造模組和假手術(shù)組,其中造模組6只,假手術(shù)組2只。同法制作模型。造模手術(shù)后,再將造模組按隨機數(shù)字表法分為1周模型組、3周模型組、6周模型組,各2只。并分別于手術(shù)后1周、3周、6周取雙側(cè)腰大肌。 對腰大肌行HE染色,觀察肌肉形態(tài)學變化；行苦味酸天狼星紅染色,觀察腰大肌內(nèi)膠原性質(zhì)及含量。 研究結(jié)果 1垂直活動度檢測： 造模后5周：模型組與假手術(shù)組相比,垂直站立紅外線計數(shù)減少,且差異有統(tǒng)計學意義(P0.05)；治療組與模型組相比,垂直站立紅外線計數(shù)增多,但差異無統(tǒng)計學意義(P0.05)。提示椎間盤損傷5周后大鼠垂直站立活動度降低。 2椎間盤HE染色： 造模后5周：髓核：假手術(shù)組呈中央密集型分布,在四周呈現(xiàn)出均勻分布狀態(tài),髓核細胞數(shù)目較多；模型組髓核組織稀疏,分布不均,伴有大量不定形物質(zhì)；治療組與假手術(shù)組相近。纖維環(huán)：假手術(shù)組可見纖維環(huán)層次分明,排列整齊,與髓核界線明顯；模型組纖維環(huán)可見纖維環(huán)排列輕度改變,有紊亂傾向；治療組纖維環(huán)形態(tài)接近假手術(shù)組。提示針刺損傷纖維環(huán)能夠造成椎間盤一定程度的損傷,點撥腰大肌治療對于椎間盤纖維環(huán)損傷可能有良性干預(yù)作用。 3椎間盤IL-1β免疫組織化學： 在纖維環(huán)細胞可檢測到椎間盤炎癥因子IL-1β的陽性表達,陽性細胞呈棕黃色。 三組間免疫組化陽性表達積分光密度(IOD)差異有統(tǒng)計學意義(P0.05)；相對于假手術(shù)組,模型組陽性表達IOD值明顯增強(P0.01)；治療組與模型組相比,陽性表達IOD值減少明顯,差異具有統(tǒng)計學意義(P0.01)。提示針刺損傷纖維環(huán)后,會造成炎性反應(yīng),且點撥腰大肌治療對椎間盤炎性改變具有良性干預(yù)作用。 4腰大肌拉伸力學： 左側(cè)腰大肌彈性模量對比：模型組彈性模量較假手術(shù)組降低,治療組較模型組增高,三組間差異無統(tǒng)計學意義(P0.05)。右側(cè)腰大肌彈性模量對比：三組間差異具有統(tǒng)計學意義(P0.05)；模型組較假手術(shù)組明顯減少,差異具有統(tǒng)計學意義(P0.01)；治療組較模型組有所增加,但差異無統(tǒng)計學意義(P0.05)。提示針刺損傷椎間盤后,大鼠右側(cè)腰大肌彈性模量出現(xiàn)一定程度的降低,點撥腰大肌治療對腰大肌彈性模量可能具有一定的干預(yù)作用。 5腰大肌HE染色： 1周腰大肌可見肌纖維有序排列,肌束緊密,其中細胞核呈橢圓形,大部分細胞核沿肌纖維的長軸邊緣排列,少數(shù)細胞核位于肌纖維內(nèi)；3周腰大肌肌纖維開始出現(xiàn)退變,肌纖維輕度腫脹,肌橫紋模糊,肌漿呈絨毛狀或融合性改變；6周肌纖維退變加重,肌橫紋模糊或消失,嗜酸性增強,肌漿絨毛狀或融合性改變明顯,肌纖維散亂無序甚至出現(xiàn)節(jié)段樣變。 6天狼星紅苦味酸染色：模型組腰大肌內(nèi)膠原含量較假手術(shù)組發(fā)生一定程度改變。 研究結(jié)論 1垂直站立紅外線計數(shù)、椎間盤形態(tài)學、椎間盤炎癥因子IL-1β及腰大肌的拉伸力學、形態(tài)學等改變能夠說明,椎間盤纖維環(huán)針刺損傷大鼠模型的腰大肌彈性模量會發(fā)生一定的改變。 2干預(yù)治療后,針刺損傷大鼠模型的腰大肌彈性模量有所恢復(fù)。推拿手法對椎間盤纖維環(huán)、髓核組織形態(tài)及炎癥因子IL-1p表達可能具有良性調(diào)節(jié)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.
【學位授予單位】：北京中醫(yī)藥大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R245
本文編號：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.
【學位授予單位】：北京中醫(yī)藥大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R245
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