發(fā)布時間：2018-07-09 23:03

  本文選題：撥法 + CCI　； 參考：《北京中醫(yī)藥大學(xué)》2017年碩士論文

【摘要】：[目的]通過行為學(xué)和形態(tài)學(xué)指標(biāo),研究撥法改善神經(jīng)病理性疼痛大鼠痛覺過敏,促進(jìn)神經(jīng)修復(fù)的可能依據(jù),通過研究促炎性因子IL-6及抗炎保護(hù)因子SOCS3在血清及脊髓中的表達(dá)情況,探究撥法治療神經(jīng)病理性疼痛,起到"消炎止痛"作用的可能起效機制。[方法]以SD雄性大鼠為實驗動物,用隨機數(shù)字表,隨機分為正常組、假手術(shù)組、模型組和撥法組,選用坐骨神經(jīng)慢性壓迫性損傷(chronic constriction injury,CCI)為模型,采用"按摩推拿手法模擬儀"(中華人民共和國發(fā)明專利號:ZL200710187403.1.)模擬撥法操作對大鼠進(jìn)行干預(yù),穴位選用殷門、承山、陽陵泉,每穴3min,每只總計9min,撥法干預(yù)每日1次,每干預(yù)10日休息1天,總共干預(yù)20天,選取造模7天后和撥法干預(yù)20天后兩個取材時間點,取材前對大鼠進(jìn)行光熱耐痛閾檢測,評價其痛覺過敏的恢復(fù)情況。對大鼠術(shù)側(cè)坐骨神經(jīng)進(jìn)行HE染色,并對結(jié)果進(jìn)行觀察分析,尋找撥法促進(jìn)神經(jīng)損傷修復(fù)的形態(tài)學(xué)證據(jù);對大鼠血清中IL-6及脊髓背角中IL-6和SOCS3的表達(dá)進(jìn)行檢測,定性定量分析指標(biāo)表達(dá)的變化,闡述撥法促進(jìn)神經(jīng)病理性疼痛緩解,起到"消炎止痛"作用的可能起效機制。[結(jié)果]1行為學(xué)結(jié)果造模7天后:正常組大鼠活動均正常;假手術(shù)組大鼠術(shù)側(cè)肢體輕微跛行,無爪攣縮,耐痛閾與正常組大鼠耐痛閾無顯著性差異(P0.05);模型組和撥法組大鼠逐漸出現(xiàn)行走時術(shù)側(cè)肢體跛行狀況,踝部下垂,且腳掌不能平放于地面,足背著地現(xiàn)象,耐痛閾與正常組相比顯著下降(P0.05)。撥法干預(yù)20天后:正常組大鼠活動均正常;假手術(shù)組大鼠輕微跛行現(xiàn)象消失,基本與正常組大鼠行動狀態(tài)無差別(P0.05);模型組大鼠疼痛癥狀未見緩解,術(shù)側(cè)持續(xù)出現(xiàn)爪攣縮,懸空不與地面接觸,關(guān)節(jié)僵硬;撥法組大鼠術(shù)側(cè)肢體仍有跋行現(xiàn)象,腳掌仍不能平放于地面,但爪攣縮現(xiàn)象較模型組有好轉(zhuǎn),光熱耐痛閾結(jié)果雖與同期正常組仍有差距(P0.05),但與同期模型組相比已有顯著性升高(P0.05)。2形態(tài)學(xué)結(jié)果造模7天后,正常組、假手術(shù)組大鼠術(shù)側(cè)坐骨神經(jīng)切片HE染色在400倍光鏡下可見坐骨神經(jīng)纖維排列整齊、緊密;模型組和撥法組則可見形態(tài)較不規(guī)則,神經(jīng)纖維排列不規(guī)則,間質(zhì)疏松水腫。撥法干預(yù)20天后,正常組、假手術(shù)組大鼠神經(jīng)切片染色狀況正常,模型組光鏡下可見軸索水腫嚴(yán)重,排列紊亂,結(jié)構(gòu)不清。撥法組可見軸索排列整齊度有所恢復(fù),神經(jīng)纖維排列有序性提高。3酶聯(lián)免疫吸附試驗(ELISA)檢測結(jié)果造模7天后:經(jīng)ELISA檢測,假手術(shù)組大鼠血清中IL-6的含量與正常組相比無顯著性差異(p0.05);模型組與同期正常組相比有顯著升高(P0.05)。撥法干預(yù)20天后:撥法組大鼠血清中IL-6的表達(dá)較同期正常組仍有顯著性差異(P0.05),但含量明顯降低,與造模7天后的同組比較,其含量約下降了 30%,逐漸接近于正常組;撥法組跟同期模型組大鼠血清中IL-6的含量相比明顯下降,有顯著性差異(P0.05)。4免疫組化染色檢測結(jié)果(1)IL-6檢測結(jié)果:造模7天后,假手術(shù)組大鼠脊髓背角中IL-6免疫陽性細(xì)胞零星分布,與正常組無顯著性差異(P0.05),模型組與同期正常組相比顯著升高(P0.05);撥法干預(yù)20天后,撥法組大鼠脊髓中IL-6的表達(dá)與同期正常組相比仍較高(P0.05),但與造模7天后的同組比較,表達(dá)量約下降了 30%;撥法組大鼠脊髓背角中IL-6的表達(dá)量約為同期模型組的70%,與同期模型組相比有顯著性差異(P0.05)。(2)SOCS3檢測結(jié)果:造模7天后,假手術(shù)組大鼠脊髓背角中SOCS3的表達(dá)與正常組相比,無顯著性差異(P0.05),模型組與同期正常組相比SOCS3的表達(dá)顯著升高(P0,05)。撥法干預(yù)20天后,撥法組大鼠脊髓背角中SOCS3的表達(dá)持續(xù)升高,約為造模7天后同組表達(dá)量的1.4倍,與同期模型組相比有顯著性差異(P0.05),約為同期模型組表達(dá)量的1.3倍。5蛋白免疫印跡檢測結(jié)果造模7天后,假手術(shù)組大鼠脊髓中IL-6的相對蛋白含量與同期正常組相比無顯著性差異(P0.05);模型組IL-6相對蛋白含量與同期正常組相比有差異性升高(P0.05)。撥法干預(yù)20天后,撥法組大鼠脊髓中IL-6的相對蛋白含量與正常組相比仍有顯著差異(P0.05),但其含量己有明顯下降;同期模型組大鼠脊髓中IL-6的相對蛋白含量雖也有下降,但比之于同期撥法組的相對蛋白含量下降程度有較大差異(P0.05)。[結(jié)論]1.撥法干預(yù)能夠提高CCI大鼠光熱耐痛閾,緩解CCI大鼠痛覺過敏的癥狀。2.撥法干預(yù)能夠促進(jìn)CCI大鼠受損神經(jīng)的再生,有利于減輕軸突腫脹,促進(jìn)軸索及神經(jīng)纖維的有序性排列。3.撥法干預(yù)能夠顯著降低血清及脊髓中促炎性因子IL-6的表達(dá),抑制外周及中樞炎性反應(yīng),起到"消炎止痛"的作用。4.撥法能夠顯著提高抗炎保護(hù)因子SOCS3在脊髓背角的表達(dá),抑制神經(jīng)炎癥反應(yīng)、中樞敏化和痛覺轉(zhuǎn)導(dǎo),緩解疼痛。本次實驗結(jié)果表明,撥法干預(yù)能夠明顯改善痛覺異常,緩解痛覺過敏,促進(jìn)受損神經(jīng)細(xì)胞的恢復(fù);撥法能夠通過下調(diào)血清及脊髓中的促炎性因子IL-6的表達(dá),以及上調(diào)脊髓中抗炎保護(hù)因子SOCS3的表達(dá)消除炎癥、抑制疼痛傳導(dǎo)、改善神經(jīng)病理性疼痛,從而起到"消炎止痛"的作用。
[Abstract]:[Objective] to study the effect of dialing method to improve the hyperalgesia of neuropathic pain rats and to promote the possibility of nerve repair through the study of behavioral and morphological indexes. Through the study of the expression of proinflammatory factor IL-6 and anti-inflammatory protective factor SOCS3 in the serum and spinal cord, we explore the effect of dialing on neuropathic pain and play the role of "anti-inflammatory and analgesic". The possible mechanism was possible. [Methods] SD male rats were used as experimental animals and randomly divided into normal group, sham operation group, model group and dialing group, chronic constriction injury (CCI) was selected as model, and "massage massage instrument Simulator" (People's Republic of China invention patent number: ZL200710) was used. 187403.1.) the rats were intervened with the simulated operation. The acupoints were selected by Yin gate, Chengshan and Yang Ling spring, each point 3min, each total of 9min, 1 times a day, each intervention was intervened for 1 days for 20 days, and two time points were selected for 7 days and 20 days after the intervention, and the photothermal pain threshold test of the rats before taking the material was evaluated and evaluated. The recovery of hyperalgesia. HE staining of the sciatic nerve in rats was carried out and the results were observed and analyzed to find morphological evidence for the repair of nerve injury by dialing method. The expression of IL-6 and SOCS3 in IL-6 and spinal dorsal horn in the rat serum was detected, the changes of the expression of qualitative and quantitative analysis index were observed, and the dialing method promoted neuropathy. The mechanism of rational pain relief played a possible effect mechanism of "anti-inflammatory and analgesic" effect. [results]1 behavior results of 7 days: normal group rats were normal, the sham operation group had slight limp, no claw contracture, and no significant difference in pain threshold between the normal group and the normal group (P0.05); the model group and the dialing group gradually appeared to walk. At the time of lame, the ankle was drooping, and the ankle was not flat on the ground, the foot was back to the ground, the pain threshold was significantly lower than the normal group (P0.05). 20 days after the intervention, the normal group rats were normal; the sham group's slight limp disappeared, and the basic group was not different from the normal group (P0.05); model group was large. The pain symptoms of the rats were not relieved, the side of the operation continued to appear claw contracture, the suspension was not exposed to the ground, and the joint was stiff. The lateral limbs of the rats in the dialing group still had a postscript, but the paws were still unable to be placed on the ground, but the contracture of the paws was better than the model group. The result of the light and heat tolerance threshold still had a gap with the normal group (P0.05), but it was with the same period model group. 7 days later than the existing significant increase (P0.05).2 morphological results, the normal group and the sciatic nerve section of the sham operation group were stained with the sciatic nerve under the 400 times light microscope, and the sciatic nerve fibers were arranged neatly and tightly under the microscope. The model group and the dialing group showed irregular shape, irregular arrangement of nerve fibers, interstitial edema and edema. The dialing method intervened for 20 days. After the normal group, the nerve section of the rats in the sham operation group was normal. The model group showed that the axon edema was serious, the arrangement was disorderly and the structure was not clear. The arrangement of axonal arrangement was recovered and the arrangement of nerve fibers was improved by the.3 enzyme linked immunosorbent assay (ELISA) for 7 days: ELISA test and sham operation There was no significant difference in the content of IL-6 in the serum of the rats compared with the normal group (P0.05); the model group was significantly higher than the normal group (P0.05). 20 days after the intervention, the expression of IL-6 in the serum of the dialing group was significantly different from that of the normal group (P0.05), but the content of the serum was significantly lower than that in the same group after the 7 days of the model. The decrease of about 30%, gradually approaching to the normal group; the dialing group was significantly lower than the IL-6 content in the serum of the model rats. There was a significant difference (P0.05).4 immunohistochemical staining results (1) IL-6 detection results: 7 days after the model, the IL-6 immunoreactive cells in the spinal dorsal horn of the sham operation group were scattered, and there was no significant difference between the normal group and the normal group. The difference (P0.05), the model group was significantly higher than the normal group in the same period (P0.05). The expression of IL-6 in the spinal cord of the dialing group was still higher than that of the normal group (P0.05) after 20 days of dialing intervention, but compared with the same group for the 7 days after the model, the expression amount decreased by 30%, and the expression of IL-6 in the spinal dorsal horn of the dialing group was about 70 in the same period model group. Compared with the model group in the same period (P0.05). (2) SOCS3 detection results: the expression of SOCS3 in the dorsal horn of the spinal cord in the sham operation group had no significant difference compared with the normal group (P0.05), and the expression of SOCS3 in the model group was significantly higher than that of the normal group (P0,05). The expression of the SOCS3 in the spinal dorsal horn of the rats in the dialing group was SO after 20 days. The expression of CS3 continued to rise, which was about 1.4 times that of the same group in the same group after 7 days of modeling (P0.05). There was no significant difference in the protein content of IL-6 in the spinal cord of the sham operation group compared with the normal group at the same time (1.3 times of the expression of.5 protein in the model group). The relative protein content of IL-6 in the model group was significantly higher than that in the normal group (P0.05). The relative protein content of the IL-6 in the spinal cord of the dialing group was significantly different from that of the normal group (P0.05) after 20 days of dialing intervention (P0.05), but the content of the protein in the spinal cord of the group was significantly decreased, although the relative protein content of the IL-6 in the spinal cord of the model group of the same stage model group also had the relative protein content. There is a significant difference in the decrease of the relative protein content in the same period group (P0.05). [conclusion]1. dial intervention can improve the light and heat tolerance threshold of CCI rats, and alleviate the symptoms of CCI rats' hyperalgesia by.2. dialing intervention can promote the regeneration of damaged nerve in CCI rats, reduce axonal swelling and promote axon and nerve fiber. The orderly arrangement of.3. dialing can significantly reduce the expression of proinflammatory factor IL-6 in the serum and spinal cord, inhibit the peripheral and central inflammatory reactions, and play the role of "anti-inflammatory and analgesic",.4. dialing can significantly improve the expression of anti-inflammatory protective factor SOCS3 in the dorsal horn of the spinal cord, inhibit the inflammatory reaction of the deity, central sensitization and pain transduction, and relieve pain. The results of this experiment showed that the dialing method can obviously improve the abnormal pain, relieve the hyperalgesia and promote the recovery of damaged nerve cells. The dialing can reduce the expression of IL-6 in serum and spinal cord, and up the expression of anti inflammatory protective factor SOCS3 in the spinal cord to eliminate inflammation, inhibit the conduction of pain and improve neuropathy. Rational pain, so as to play the role of "anti-inflammatory and analgesic".
【學(xué)位授予單位】：北京中醫(yī)藥大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R244.1;R-332

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