摘要
目的探讨推拿康复治疗痉挛型小儿脑性瘫痪(cerebral palsy,CP)的机制。方法选择30例痉挛型小儿CP患者采用选择性脊柱推拿康复疗法进行治疗,同时设立10名性别和年龄匹配的健康志愿者作为对照组。使用统一的观察量表观察病人的肌痉挛程度、关节活动度和7项疗效评估情况,疗程结束后进行统计分析。分别采集治疗前、治疗3m后各CP患者和健康者的外周血3mL,用密度梯度离心法分离外周血单个核细胞,应用炎症细胞因子与受体PCR芯片进行基因表达检测,绘制基因表达谱,筛选有表达差异的基因。外周血离心取血清,运用酶联免疫吸附法(ELISA)检测血清中炎性细胞因子TNF-α、IL-6和IL-10蛋白表达。结果治疗后患儿肢体的肌痉挛程度和关节活动度明显改善,与治疗前相比差异具有统计学意义(P<0.05),7项疗效评估有效率为90.00%,表明推拿康复疗法对CP取得较好的疗效。基因芯片检测结果表明,与健康对照组相比,治疗前CP患者组表达上调的促炎性反应细胞因子与受体基因有7个,表达下调的抗炎细胞因子与受体基因有6个。与治疗前比较,治疗3m后CP患者组中表达上调的抗炎细胞因子与受体基因有4个,下调的促炎细胞因子与受体基因有6个。ELISA检测结果发现,治疗前CP患者组外周血血清中TNF-α和IL-6蛋白水平都显著高于健康对照组(P<0.01),而IL-10蛋白水平都显著低于健康对照组(P<0.05);与治疗前比较,治疗3m后CP患者组血清中TNF-α和IL-6蛋白水平都明显降低,IL-10蛋白水平显著升高(P<0.05)。与健康对照组相比,治疗前CP患者组血清中TNF-α/IL-10和IL-6/IL-10比值均显著升高(P<0.01);与治疗前比较,治疗3m后CP患者组血清中TNF-α/IL-10和IL-6/IL-10比值均显著降低(P<0.05)。结论 CP患者存在促炎性细胞因子基因表达上调和抗炎症细胞因子基因表达下调,这可能是CP患者机体中出现高促炎性反应状态的原因和发病的新机制;推拿康复疗法可能通过下调促炎性细胞因子基因表达和上调抗炎性细胞因子基因表达,重塑促-抗炎性细胞因子网络和促-抗炎性反应体系新的平衡来治疗痉挛型小儿脑瘫。
Objective To investigate the mechanisms of massage rehabilitation therapy for cerebral palsy（CP）. Methods 30 cases with CP were treated with the selected spinal manipulative therapy（SSMT）for 3 months,while 10 healthy age-and sex-matched children were regarded as controls. Physical extent of muscle cramps,joint activity, and 7 items of therapeutic effective evaluation were measured by the same measuring scale. Peripheral blood（3 mL） was obtained from every CP patients and controls. Gene expression profiles of the peripheral blood mononuclear cells （PBMCs） ,which were isolated from the peripheral by Flcoll density gradient centrifugation from the CP cases before and after treatment,as well as the controls, were analyzed by using the human inflammatory cytokines ＆ receptors PCR array,respectively. Differentially expressed genes were screened. At the same time, the levels of TNF-a, IL-6 and IL-10 in the peripheral blood were evaluated by Enzyme Linked Immunosorbent Assay（ELISA）. Results Compared with pre treatment, the physical extent of muscle cramps and joint activity were improved（P〈0.05）. The effective rate was 90.00% in post-treatment,indicating that the SSMT was a new effective therapeutic method for CP. Compared with the healthy control group, there were 7 differentially up regulated expressed pro-inflammatory genes（P〈0.05） ,while 6 down-regulated expressed anti-inflammatory genes in in the pre-treatment CP group（P〈 0.05）. Compared with the pre-treatment CP group, there were 4 differentially up-regulated expressed anti inflammatory genes, while 6 down-regulated expressed pro-inflammatory genes in the post-treatment CP group. Compared with the healthy control group,the protein levels of TNF-a and IL-6 in the PBMCs in the pre-treatment CP group increased significantly（P〈0.01 ）, while the level of IL-10 decreased （P〈0.05）. Compared with the pre treatment CP,the levels of proteins of TNF-a and IL-6 in the PBMCs in the post-treatment CP group were decreased significantly（P〈0.01） ,while the level of IL-10 was increased（P〈0.05）. The ratios of TNF-a to IL-10 and IL 6 to IL-10 in the pre-treatment CP group increased significantly compared with those in the healthy control group（P〈0. 01）,which were reversed by SSMT in the post treatment CP group（P〈0.05）. Conclusions Gene expressions of pro-inflammatory cytokines and receptors were up regulated in the PBMCs of the CP cases, which may eventually contribute to disequilibrium of the network of pro-and anti-inflammatory cytokines and receptors and disequilibrium of response of pro-and anti inflammatory system and high inflammatory response state, thus providing new mechanistic insight into CP. The SSMT is an effective therapeutic method for CP and its mechanism involves to rebuild the equilibrium of inflammatory response state.
出处
《成都医学院学报》
CAS
2012年第4期527-532,共6页
Journal of Chengdu Medical College
基金
国家自然科学基金(81102541)
国家十二五支撑课题(2013BAI01B003)
国家中医药管理局"十二五"重点学科中医儿科学项目
关键词
脑性瘫痪
选择性脊柱推拿
康复
单个核细胞
基因芯片
炎症细胞因子与受体
酶联免疫吸附
Cerebral palsy
Selected spinal manipulative therapy
Rehabilitation
Mononuclear cell
Gene chip
Inflammatory cytokins and receptors
Enzyme Linked Immunosorbent Assay
