电针对甲状腺区切口痛大鼠颈段脊髓胶质细胞活性的影响

目的:观察电针(electroacupuncture,EA)不同穴位对甲状腺区切口痛大鼠颈段脊髓小胶质细胞和星形胶质细胞活性的影响,探讨针刺镇痛行甲状腺手术的机制.方法:Wistar雄性大鼠60只随机分为正常组、模型组、扶突组、合谷-内关组、足三里-阳陵泉组,每组12只.除正常组外,余组沿大鼠颈中线做长约1.5 cm的纵行切口,制作甲状腺区切口痛模型.扶突组、合谷-内关组、足三里-阳陵泉组于造模后4h、24 h、48 h分别电针双侧“扶突”、“合谷”“内关”穴、“足三里”“阳陵泉”穴,1次/d,连续3 d;正常组、模型组不予其他处理.采用热辐射测痛仪测量大鼠切口部位热痛阈;用荧光定量RT-PCR、蛋白免疫印迹法(WB)分别检测各组大鼠干预后颈段(C2-C6)脊髓小胶质细胞活化标记物(Iba1、CD11b)及星形胶质细胞特异性标记物胶质纤维酸性蛋白(glial fibrillary acid protein,GFAP)基因及蛋白的表达.结果:干预结束后,与正常组相比,模型组大鼠颈部的热痛阈值明显降低(P＜0.05),C2-C6脊髓内Iba1、CD11b及GFAP mRNA与蛋白表达均明显上调(P＜0.05,P＜0.01);与模型组相比,扶突组和合谷-内关组大鼠的热痛阈值显著升高(均P＜0.05),足三里-阳陵泉组大鼠的热痛阈值变化不明显(P＞0.05),扶突组Iba1、CD11b、GFAP基因和蛋白的表达较模型组降低(均P＜0.05),合谷-内关组大鼠脊髓中Iba1 mRNA、CD11b蛋白、GFAP mRNA和蛋白表达明显低于模型组(均P＜0.05),足三里-阳陵泉组脊髓Iba1、CD11b和GFAP蛋白与模型组相比差异无统计学意义(均P＞0.05);足三里-阳陵泉组大鼠C2-C6颈段脊髓内Iba1 mRNA、CD11b mRNA与蛋白表达均明显高于扶突组(P＜0.01,P＜0.05),Ibal mRNA、CD11b蛋白表达水平高于合谷-内关纽(P＜0.05),GFAP mRNA与蛋白表达均明显高于扶突穴组与合谷-内关组(均P＜0.05).结论:电针“扶突”或“合谷”“内关”可缓解大鼠颈部急性切口痛,该作用可能与其下调脊髓内小胶质细胞和星形胶质细胞的活性密切相关.

Objective:To explore the relationship of peripheral nerve ultrastructure and its associated pro-tein expression in experimental autoimmune neuritis ( EAN) . Methods:EAN was established in Lewis rats using an emulsified mixture of P0 peptide 180-199,Mycobacterium tuberculosis,and incomplete Freund's adjuvant. Rats given saline solution were used as a control group. Sciatic nerve ultrastructure and immunofluorescence histopathol-ogy were measured at the neuromuscular severity peak on day 18 post-induction. Cell-specific protein markers were used for immunofluorescence histopathology staining to characterize sciatic nerve cells:CD3 ( T cell) , Iba-1 ( mi-croglia), S100(myelin), and neurofilament 200(axon). Results:The results showed that swelling of the myelin lamellae, vesicular disorganization,separation of the myelin lamellae, and an attenuation or disappearance of the axon were observed by transmission electron microscopy in the EAN group. CD3 and Iba-1 increased significantly in the structures characterized by separation or swelling of the myelin lamellae,and increased slightly in the struc-tures characterized by vesicular of the myelin lamellae, S100 decreased in the structures characterized by vesicular disorganization or separation of the myelin lamellae. And neurofilament 200 decreased in the structures character-ized by separation of the myelin lamellae. Moreover,we found that Iba1 were positive in the myelin sheath, and o-verlapped with S100 , which significantly indicated that Schwann cells may play as macrophage-like cells during the disease progression of ENA. Our findings may be a significant supplement for the knowledge of EAN model, and may offer a novel sight on the treatment of GBS.

脉冲射频背根神经节对CCI模型大鼠脊髓中 Iba1和TRPV1表达的影响

目的:对慢性坐骨神经缩窄损伤(chronic constriction injury,CCI)大鼠实施背根神经节(dorsal root ganglion,DRG)脉冲射频(pulsed radiofrequency,PRF),探讨对其疼痛行为学和脊髓中离子钙结合适配器分子1(ionized calcium binding adapter molecule 1,Iba1)、瞬时感受器电位香草酸受体1(transient receptor potential vanilloid type 1,TRPV1)表达的影响.方法:雄性SD大鼠100只随机分为4组:假手术组(Sham)、CCI组、CCI+PRF组、CCI+free-PRF组,每组25只.除Sham外,其余三组结扎右侧坐骨神经主干制作CCI模型.CCI后第7 d,CCI+PRF组于术侧L4-5 DRG行6 min的标准PRF治疗.CCI+free-PRF组仅在相同位置放置射频电极,无脉冲治疗.观察大鼠CCI前,CCI后1、3、5、7 d,PRF后1、7、14 d痛阈改变.术前、CCI后第7 d、PRF后14 d采用RT-PCR、免疫荧光法检测脊髓Iba1(小胶质细胞特异性抗体)和TRPV1表达.结果:CCI后,与Sham组比,CCI组、CCI+PRF组、CCI+free-PRF组大鼠痛阈均显著降低(P<0.05),脊髓Iba1、TRPV1表达均增强(P<0.05).脉冲射频后,CCI+PRF组痛阈较CCI组和CCI+free-PRF组显著升高(P<0.05),脊髓Iba1、TRPV1表达则明显减少(P<0.05).结论:脉冲射频可有效减轻CCI大鼠的神经病理性疼痛,可能与抑制大鼠脊髓中小胶质细胞活化、TRPV1表达有关.

金银花水提物对小鼠糖尿病视网膜病的改善作用

目的：观察金银花水提物（Lonicerae Japonicae Flos a-queous extract，FL）对糖尿病视网膜病（diabetic retinopathy， DR）的改善作用及其机制。方法采用链脲佐菌素（strep-tozotocin，STZ）腹腔注射制备 C57小鼠糖尿病模型，给药2个月后形成了 DR 早期病程阶段即非增殖性糖尿病视网膜（non-proliferative DR，NPDR）。使用伊文氏蓝实验检测血－视网膜屏障（blood-retinal barrier，BRB）的渗漏情况；运用Western blot 实验检测相关蛋白的表达；酶联免疫实验（En-zyme-linked immunosorbent assay，ELISA）检测血清中炎性因子的表达。结果伊文氏蓝实验发现 FL 可以剂量依赖性的抑制 STZ 诱导糖尿病小鼠中发生的 BRB 渗漏。Western blot结果显示 FL 可以抑制 STZ 诱导糖尿病小鼠视网膜组织中核因子κB （nuclear factor-κB，NF-κB）p65亚单位的转核激活，FL 还可以抑制视网膜组织中 kappa B 抑制剂（inhibitor of κB，IκB）、抑制性κB 激酶（inhibitor of nuclear factor κB kinase，IKK）、p65的磷酸化激活。进一步研究发现 FL 可以抑制 STZ 诱导糖尿病小鼠视网膜组织中升高的小胶质细胞／巨噬细胞特异性蛋白（ionized calcium binding adapter mole-cule 1，Iba1）的表达，提示 FL 可以抑制神经小胶质细胞的活化。ELISA 实验结果发现 FL 能降低 STZ 诱导糖尿病小鼠血清中升高的炎性细胞因子如白介素（interleukin，IL）-6，-1β。结论FL 可能通过抑制神经小胶质细胞的活化，抑制NF-κB 介导的炎性信号通路，缓解视网膜的炎性损伤和 BRB的渗漏，从而发挥改善 DR 的药效活性。

慢性肾衰竭大鼠小胶质细胞数量及形态变化

目的:观察慢性肾衰竭(CRF)大鼠脑内小胶质细胞数量及形态变化,探讨CRF大鼠的脑内炎症反应,为尿毒症脑病的发病机制提供新的思路.方法:应用5/6肾大部分切除术建立CRF模型,将40只雄性SD大鼠随机分为模型组(CRF组,n=20)和对照组(假手术组,n=20).术后10周,测定两大鼠血肌酐、尿素、24h尿蛋白及尿量;并采用免疫组化检测两大鼠小胶质细胞的数量及形态变化.结果:CRF组血肌酐、尿素氮及24 h尿蛋白定量均较对照组升高4～5倍,差异有统计学意义(P＜0.01)；CRF组大鼠小胶质细胞数量明显增多,胞体变大,突起粗短,与对照组比较,差异有显著性意义(P＜0.01).结论:CRF模型大鼠小胶质细胞被激活,存在脑内炎症反应,可能与UE发病机制相关.