【作者】 刘威；

【导师】 刘忠良；

【作者基本信息】 吉林大学 ， 康复医学与理疗学， 2021， 硕士

【摘要】 复杂性区域疼痛综合征Ⅰ型（complex regional pain syndrome Ⅰ,CRPSⅠ）常常发生于损伤之后,以异常性疼痛为主要的临床特点,可伴有不同程度的感觉障碍、运动障碍及自主神经功能障碍。急性期（温暖型）可表现为明显的红、肿、热、痛等炎症反应,在此期中,局部组织可表现血管扩张、炎性介质释放等。一部分人经过一段时间后可自愈,或经一段时间的治疗后可治愈。但也有一部分人病程迁延不愈,逐渐转为慢性期（寒冷型）,此期CRPSⅠ患者炎症反应逐渐减退,表现为难以忍受的疼痛症状,且该症状持续不缓解,治愈率低,严重影响人们的生活质量,甚至患者会出现心理问题。该病的诊断标准目前以临床症状为主,并没有明确的实验室及影像学等客观证据的支撑。治疗方法也比较局限,因此,我们寻求一种新的治疗方法缓解此征的症状和体征。目的:1.探究不同频率（1Hz、10Hz）的重复外周磁刺激（repetitive peripheral magnetic stimulation,rPMS）对胫骨骨折外固定引起的CRPS Ⅰ大鼠模型是否具有治疗作用。2.探究不同频率（1Hz、10Hz）的rPMS治疗胫骨骨折外固定引起的CRPSⅠ大鼠模型后血清中白介素6（interleukin 6,IL-6）、肿瘤坏死因子α（tumor necrosis factorα,TNFα）、神经元特异性烯醇化酶（neuron-specific enolase,NSE）的变化。3.探究不同频率（1Hz、10Hz）的rPMS治疗胫骨骨折外固定引起的CRPS Ⅰ大鼠患侧L_4-5背根神经节（dorsal root ganglia,DRG）内神经生长因子（nerve growth factors,NGF）的基因表达量及蛋白定量的变化。方法:本论文包括三部分内容。第一部分中,选用成年雄性Sprague-Dawley大鼠（1月龄）,随机分成未建模的A组（空白组）6只和建模组B组（模型对照组）6只、C组（CRPS Ⅰ+1Hz组）6只、D组（CRPS Ⅰ+10Hz组）6只,备用3只,共27只。其中21只建模组大鼠建立胫骨骨折外固定模型,建模成功后剔除掉建模失败的大鼠（死亡、患肢坏死者）,备用组大鼠随机分到B、C、D组中,最终A组6只,B组5只和C组6只,D组6只。建模前及建模后进行体重、机械性刺激缩足阈值（paw mechanical withdrawal threshold,PMWT）和热缩足反射潜伏期（paw thermal withdrawal latency,PTWL）评估大鼠胫骨骨折外固定模型的成功与否及骨折后大鼠热痛及机械痛情况。分别用1Hz和10Hz的rPMS治疗C组和D组的大鼠,治疗4周后再次进行PMWT和PTWL的测定,以评估CRPS Ⅰ大鼠的治疗效果。第二部分中,提取治疗前后的CRPS Ⅰ大鼠血清并用酶联免疫吸附法（enzyme-linked immunosorbent assays,ELⅠSA）进行IL-6、TNFα、NSE的浓度,以评估rPMS治疗后的大鼠血清中IL-6、TNFα、NSE的变化。第三部分中,提取治疗后的CRPS Ⅰ大鼠L_4-5 DRG,并用实时聚合酶链锁反应（real-time polymerase chain reaction,RT-PCR）和蛋白质印迹法（western-blot）测定DRG中NGF基因表达量及蛋白定量情况,以评估rPMS治疗后大鼠DRG中NGF的表达情况。结果:第一部分中,治疗后对所有建模大鼠患侧后足进行PTWL、PWMT的测定,统计结果显示治疗后B组、C组、D组大鼠患侧后足PTWL均较治疗前增加,且经过治疗的C、D组治疗效果优于未治疗的B组,差异有统计学意义（P<0.05）。治疗后C、D组大鼠患足PWMT较治疗前增加（P<0.05）,且D组治疗效果优于B组,差异有统计学意义（P<0.05）。第二部分中,治疗前及治疗后均对所有大鼠血清进行IL-6、TNFα、NSE的测定,发现治疗后B组、C组、D组大鼠血清中IL-6均较治疗前下降（P<0.05）,治疗后D组低于B组（P<0.05）,C、D组未见明显差异;治疗后C、D组大鼠血清中TNFα较治疗前下降（P<0.01）,且C、D组低于B组（P<0.05）,C、D组未见明显差异;治疗后B组、C组、D组大鼠血清中NSE均较治疗前下降（P<0.05）,但三组之间治疗后浓度无明显差异。第三部分中,治疗后取大鼠患侧L_4-5DRG进行NGF表达的测定,C组、D组大鼠DRG中NGF基因表达量及蛋白定量均明显少于B组（P<0.01）,C、D组之间未见明显差异。结论:1.高频与低频rPMS均可以提高CRPSⅠ大鼠热痛、机械痛阈值,缓解大鼠疼痛症状,高频与低频rPMS之间无差异。2.高频与低频rPMS可减少CRPS Ⅰ大鼠血清中的IL-6、TNFα浓度,减轻炎症反应,缓解CRPS Ⅰ疼痛,高频与低频rPMS之间无差异。3.高频与低频rPMS可以减少CRPS Ⅰ大鼠DRG中的NGF表达,修复DRG功能,缓解疼痛,高频与低频rPMS之间无差异。更多还原

【Abstract】 CRPS Ⅰ often occurs after injury,intractable pain as the main clinical features,may be accompanied by varying degrees of sensory abnormalities,movement disorders,and autonomic dysfunction.The acute phase（warm type）can be marked by inflammation signs such as redness,swelling,heat and pain.In the meantime,local tissues can present vascular dilation,inflammatory media exudation and so on.Some people can heal themselves over a period of time,or they can be cured after a period of treatment.But there are also some illnesses persistent cured,gradually into the chronic phase（cold type）,the period of CRPS Ⅰ inflammation in patients with gradually decreased,present obvious symptoms of unbearable pain and symptoms do not ease,cure rate is low,the serious influence on the quality of peoples’lives,even introduced mental symptoms.At present,the diagnostic criteria of this disease are rarely based on clinical practice,which are mainly based on clinical symptoms without clear support of objective evidence such as laboratory and imaging.Treatments are also limited.So we are looking for a new treatment to relieve the pain of these patients.Objective:1.To investigate whether rPMS with different frequencies（1Hz,10Hz）has therapeutic effect on tibial fracture rats.2.To investigate the changes of IL-6,TNFαand NSE in serum of CRPS Ⅰ rats after treatment of tibia fracture with rPMS of different frequencies（1Hz and 10Hz）.3.To investigate the changes of NGF gene expression and protein quantification in L_4-5DRG of the affected side of the rats after the treatment of tibia fracture with rPMS of different frequencies（1Hz,10Hz）in CRPS Ⅰ rats.Methods:This thesis includes three parts.In the first part,adult male Sprague-Dawley rats（1 month old）were selected and randomly divided into unmodulated group A（blank group）6 rats,modulated group B（model control group）6 rats,C（CRPS Ⅰ+1Hz group）6 rats,and D（CRPS Ⅰ+10Hz group）6 rats with 3 spare rats,with A total of 27 rats.Among them,21 rats of the model groups were established external fixation model of tibial fracture.After successful modeling,rats in the failure model group（dead,damaged limb）were excluded.Rats in the reserve group were randomly divided into group B,C and D,and finally 6 rats in group A,5 rats in group B,6 rats in group C and6 rats in group D.Paw Mechanical Withdrawal Threshold（PMWT）and Paw Thermal Withdrawal Latency（PTWL）were measured before and after modeling to evaluate the success of the external fixation model of tibial fracture in rats and the conditions of thermal pain and mechanical pain after fracture.Results:In the first part,PTWL and PWMT were measured for the affected posterior foot of all modeling rats after treatment,and the statistical results showed that PTWL of the affected posterior foot of rats in group B,C and D increased after treatment compared with before treatment,and the treatment effect of the treated group C and D was better than that of the untreated group B,with statistical significance（P<0.05）.After treatment,the PWMT of rats in group C and D were increased compared with that before treatment（P<0.05）,and the treatment effect of group D was better than that of group B,the difference was statistically significant（P<0.05）.In the second part,IL-6,TNFαand NSE of all rats were measured before and after treatment.Ⅰt was found that IL-6 of rats in group B,C and D decreased after treatment（P<0.05）,and there was a difference between group D and group B after treatment（P<0.05）,but no significant difference was found between group C and D.After treatment,the serum level of TNFαin group C and D decreased compared with that before treatment（P<0.01）,and there was a difference between group C and D and B（P<0.05）,but no significant difference was found between group C and D.After treatment,the NSE levels in group B,C and D were all decreased compared with before treatment（P<0.05）,but there was no significant difference between the three groups after treatment.In the third part,after treatment,DRG of L_4-5of the affected side of rats were taken for the determination of NGF expression.The NGF gene expression and protein quantity in the DRG of rats in group C and D were significantly lower than those in group B（P<0.01）,and there was no significant difference between group C and D.Conclusion:1.Both high-frequency and low-frequency rPMS can improve the heat pain and mechanical pain thresholds of CRPSⅠrats,and relieve the pain symptoms of rats.There is no difference between high-frequency and low-frequency rPMS.2.High-frequency and low-frequency rPMS can reduce IL-6 and TNFαof serum in CRPS Ⅰ rats,and reduce inflammatory response and alleviate allodynia,and there is no difference between high-frequency and low-frequency rPMS.3.High frequency RPMS and low frequency rPMS can reduce NGF expression in DRG of CRPS Ⅰ rats,repair DRG function and relieve allodynia,and there is no difference between high and low frequency rPMS.更多还原