Abstract As stimulation points in acupuncture,acupoints have unique local pathological characteristics. Acupoint sensitization is the transformation of an acupoint from a ‘silent state’ to an‘activ ated state’ and the specificity of sensitization is important for disease investigation, and its dynamics vary significantly with the degree of sensitization.The mechanism of action of l ocal acupoints on the skin needs to be further clarified to provide reference value for sub sequent acupuncture treatment.To explore the degree of acupoint sensitization relatedto the and the underlying mechanism of local content of small biomolecules, mainly mastcells.We established a knee osteoarthritis (KOA) model by intra-articular injection of sodiumiodoac etate (MIA) into the left knee of rats.The distal points of the extremities (Left Tianjing,Ri ght Tianjing,Left Heding,Right Heding)were selected for the study.The mechanical pain thre shold at local acupoints was detected by Electric Von Frey, and the aggregation andthreshi ng status of mast cells(MCs) were detected by toluidine blue staining.The MCs aggregatio n and degranulation status of each group were detected by toluidine blue staining;the chan ges in the content of trypsin, 5-HT and HA indexes at different activation points weredete cted by immunofluorescence double-labeling method.It showed that the content of trypsin, 5-HT and HA at different activated acupuncture points varied more than the resting state of the local acupuncture points, with the right Tianjing acupoint having a higher content t han the other points, indicating that the number of MCs and the content of trypsin, 5-HT and HA produced by degranulation may be one of the cellular mechanisms occurring dur ing acupuncture point sensitization. The pain thresholds of different acupoints were signific antly different, and the sensitization degree was in the order of right Tianjing acupoint（SJ 10）>right Heding acupoint（EX-LE2）>left Heding acupoint（EX-LE2）>left Tianjing acu point（SJ10）.It indicated that the sensitization differences clarify the degrees of variationw ith the differences in point selection,which helped to broaden the understanding ofthe char acteristics of local points and explore the basic pathology of localpoints and provided som e theoretical basis for clinical intervention selection.

Objective To elucidate the differences in manual acupuncture effectiveness at sensitized points by investigating the mechanisms of local skin action at different sensitization points in rats with knee osteoarthritis (KOA). Methods Forty Sprague–Dawley rats were equally divided into control, model (1 mg of monoiodoacetate into the right knee joint cavity), sham operation, manual acupuncture at right Tianjing acupoint (MAR-SJ 10), and left SJ 10 groups. Safranine-O and fast green staining were used to assess the modeling. The morphological and functional changes in mast cells (MCs) were assessed during acupoint sensitization using toluidine blue and immunofluorescence staining. The levels of serotonin, histamine, substance P (SP), and tryptase at skin acupoints and serum levels of IL-β, IL-6, and TNF-α were detected using ELISA. Results After 14 days of treatment, the number of MCs and their degranulation rates were statistically higher in the model group than in the control group (both P < .001). After applying acupuncture, the levels of 5-HT, HA, and SP at skin acupoints were lower than those in the model group (all P < .05), and tryptase level was higher (both P < .05). Tryptase level was higher on the skin at the MAL-SJ 10 acupoint than that on the MAR-SJ 10 acupoint (P = .004). Compared with the model group, the serum levels of IL-1β, IL-6, and TNF-α in the MAR-SJ 10 and MAL-SJ 10 groups were lower (all P < .05). Conclusion Acupuncture at KOA-sensitized acupoints mitigates joint injury in KOA rats and may bidirectionally regulate local MCs of these acupoints. This finding not only enhances the reference value of sensitizing points in clinical diagnosis and treatment, but also contributes to the understanding of the biological mechanisms underlying acupuncture intervention at sensitizing points.

电针治疗脊髓损伤可以显著提高脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)、原肌球蛋白受体激酶B(tropomyosin-receptor kinase,TrkB)蛋白的表达,下调 p75 神经营养素受体(p75 neurotrophin receptor,p75 NTR)蛋白的表达.电针可以通过提高BDNF促进神经元的存活、促进受损纤维的再生、促进神经可塑性、促进髓鞘形成BDNF基因修饰的成纤维细胞.BDNF诱导的TrkB信号可能通过四种主要的细胞内途径影响突触可塑性、轴突生长、存活和细胞内阳离子平衡:(1)经磷脂酰-3激酶(P1-3K)/Akt通路促进神经元树突分枝和树突棘生成,影响突触生长和结构形成.(2)受体诱导小分子GTP酶Ras的激活,经过一系列复杂的磷酸化过程,最终激活许多细胞外信号调节激酶(extracellular signal-regulated Kinase,ERK)途径并对环腺苷3',5'-单磷酸含量进行调节促进轴突生长.(3)经磷脂酶C-γ(PhospholipaseC-y,PLC-γ)/肌醇3-磷酸(inositol triphosphate,IP-3)通路参与Ca2+的调控,促进Ca2+从细胞内储存释放,促进突触可塑性等.(4)此外通过N-甲基-D-天冬氨酸(N-Methyl-D-aspartic acid,NMDA)受体的磷酸化,TrkB信号可能导致钙和钠内流的增加.故电针治疗脊髓损伤,可使BDNF与TrkB高亲和力结合,促进脊髓损伤的修复.其可能是通过激活BDNF-TrkB信号通路,抑制BDNF前体(brain-derived neurotrophic factor precursor,proBDNF)-p75NTR信号通路来实现对神经的再生修复.

OBJECTIVE:To observe the effect of electroacupuncture(EA) on activities of A2 type astrocytes(A2s)and A1 type astrocytes (A1s) , expressions of neurofilament protein 200 (NF-200, a marker of axon regeneration), nexin 1(NL1, a marker of synaptic regeneration), and regeneration of Nissl bodies in rats with spinal cord injury (SCI), so as to explore its mechanisms underlying improvement of SCI.METHODS:A total of 75 male SD rats were rando-mized into sham operation, model, antibody neutralizing (AN), EA and EA+AN groups, with 15 rats in each group. The SCI model was established by using an infinite field impactor to deliver an about 200 k dyne weight onto the exposed spinal cord after making a dorsal laminectomy at vertebral level T10. EA (2 Hz, 1 mA) was applied to"Dazhui"(GV14) and "Mingmen"(GV4) for 20 min, once daily for 28 days. After modeling, intraspinal injection of neutralizing antibodies IL-1α, TNF-α and complement 1q (C1q, 2 μL) to the injured spinal locus for inhibition of A1 type astrocytes (A1s) was conducted on the 1st, 7th , 14th and 21st day for rats of AN and EA+AN groups. BBB rating scale was used to evaluate hindlimb locomotor function on day 1, 7, 14, 21 and 28 after modeling. The activation of A2s (its specific marker S100a10), astrocyte (its specific marker glial fibrillary acidic protein, GFAP), and A1s (its specific marker C3) in the spinal cord was detected by immunofluorescence, and the protein expressions of NF-200 and NL1 in the spinal cord detected by Western blot and immunohistochemistry, separately, and the neuronal regeneration was observed after Nissl staining.RESULTS:After SCI, the BBB scores at 1 , 7, 14, 21 and 28 day, and the immunoactivity of NL1 and NF-200 were significantly decreased (P<0.01), and the fluorescence intensity of double labelled S100a10 (A2s)/GFAP and C3, and the expression of NF-200 were considerably increased in the model group (P<0.05, P<0.01). In contrast to the model group, the BBB scores at 7, 14, 21 and 28 day, and the immunoactivity of NL1 and NF-200, and the fluorescence intensity of A2s/GFAP in the AN, EA and AN+EA groups, and the expressions of NL1 in the EA and AN+EA groups, and expression of NF-200 protein in the AN+EA group were evidently increased (P<0.05, P<0.01), and the fluorescence intensity of C3 was strikingly decreased in the EA group (P<0.01). The effect of AN+EA was significantly superior to that of single AN and EA in increasing BBB scores at 14, 21 and 28 day, and in up-regulating the immunoactivity of NF-200(P<0.01, P<0.05). Nissl staining showed damaged structure of the gray matter of the spinal cord, atrophy of the Nissl body, and pyknosis of neurons, which was milder in the AN and EA groups, particularly in the AN+EA group.CONCLUSION:EA at GV14 and GV4 may promote activation of A2s and promote regeneration of axons and synapses in SCI model rats.

目的:观察电针联合rTMS对脊髓损伤(SCI)大鼠损伤脊髓NGF、NT-3蛋白表达的影响.方法:SD大鼠120只随机分为空白组、假手术组、模型组、电针组、rTMS组与联合组(电针+rTMS),其中模型组、电针组、rTMS组和联合组再随机分为1 d组与7 d组,各12只.电针组给予电针治疗,rTMS组给予rTMS治疗,联合组则先进行rTMS治疗后再进行电针治疗.采用BBB评分观察治疗前后运动功能的改善程度,HE、尼氏染色观察神经元恢复情况,Westernblot检测NGF、NT-3蛋白表达的情况.结果:造模后模型和各干预组NGF、NT-3含量下降,治疗7 d后,各干预组与模型组比较NGF、NT-3增加(P＜0.05或P＜0.01),电针组与rTMS组比较NGF含量增加(P＜0.01),联合组与rTMS组比较NGF含量增加(P＜0.01),联合组与电针组比较NGF含量增加(P＜0.05).rTMS组与模型组比较NT-3含量增加,差异无统计学意义(P＞0.05),电针组、联合组与模型组比较NT-3含量增加(P＜0.01),电针组与rTMS组比较NT-3含量增加,差异无统计学意义(P＞0.05),联合组与rTMS组比较NT-3含量,差异无统计学意义增加(P＜0.01),联合组与电针组相比NT-3含量增加(P＜0.05).结论:电针联合rTMS能促进NGF、NT-3蛋白的表达,促进SCI的恢复.

目的:观察电针联合重复经颅磁刺激对脊髓损伤(SCI)大鼠行为学、神经营养因子BDNF及其受体TrkB和p75NTR表达的影响,探索电针联合重复经颅磁刺激治疗SCI的可能机制.方法:将120只清洁级SD大鼠随机分为空白组(12只)、假手术组(12只)、模型组(24只)、电针组(24只)、经颅磁刺激组(24只)和联合组(24只).模型组、电针组、经颅磁刺激组及联合组再按干预1 d、7 d分为2个亚组,每个亚组12只.并采用改良式Allen's打击装置制备大鼠SCI模型.空白组不做任何处理;假手术组只暴露脊髓,不打击;电针组穴取"大椎"和"命门";经颅磁刺激组应用90％静息运动阈值;联合组用电针加经颅磁刺激治疗.1 d组于造模清醒后治疗1次,7 d组于1 d组同一时间每天重复治疗1次.采用BBB评分评定大鼠脊髓损伤后的功能;HE染色法观察脊髓组织病理形态改变;免疫组织化学法、Western blot法检测脊髓组织BDNF、TrkB及p75NTR蛋白表达.结果:BBB评分结果显示,与空白组比较,1 d、7 d模型组BBB评分均显著下降,差异具有统计学意义(P＜0.05);7 d时:与模型组和经颅磁刺激组比较,电针组和联合组BBB评分显著升高,差异具有统计学意义(P＜0.05);与电针组比较,联合组BBB评分显著升高,差异具有统计学意义(P＜0.05).在HE染色中,干预1 d,可见模型组及各干预组脊髓组织结构被破坏,组织坏死,分界不清,中央管结构紊乱,红细胞大量聚集明显的空泡等;干预7 d,各组出血减少,电针组、联合组较模型组组织结构更加完整、空洞减少.免疫组织化学法及Western blot法结果显示,与空白组比较,1 d、7 d模型组BDNF、TrkB蛋白表达均显著下降,差异具有统计学意义(P＜0.05);7 d时:与模型组和经颅磁刺激组比较,电针组和联合组大鼠脊髓组织中BDNF、TrkB蛋白表达显著升高,差异具有统计学意义(P＜0.05);与电针组比较,联合组大鼠脊髓组织中BDNF、TrkB蛋白表达显著升高,差异具有统计学意义(P＜0.05).与空白组比较,1 d、7 d模型组p75NTR蛋白表达量均显著上升,差异具有统计学意义(P＜0.05);7 d时:与模型组和经颅磁刺激组比较,电针组和联合组p75NTR蛋白表达显著下降,差异具有统计学意义(P＜0.05);与电针组比较,联合组p75NTR蛋白表达显著下降,差异具有统计学意义(P＜0.05).结论:督脉电针联合重复经颅磁刺激可改善脊髓损伤大鼠神经功能损伤症状,其机制可能与上调脊髓中BDNF、TrkB以及下调了 p75NTR的表达有关.电针相对单纯经颅磁刺激在大鼠早期干预中效果更好,联合治疗对比单纯电针的效果更显著.

痛情绪是指因疼痛引发的情绪和情感体验,是疾病过程中最常见的一种情绪.痛情绪相关神经机制非常复杂,但主要与单胺类神经递质、神经肽和某些神经环路有关,笔者将结合目前研究现状分别从以上两方面展开,就痛情绪相关单胺类神经递质和神经肽在受体分类、脑区通路、共疾病以及各神经递质之间的联系和痛情绪相关神经环路中各个蛋白的作用机制等方面进行探讨.

随着近年来交通和工业的快速发展,脊髓损伤患者也不断增加,现已成为严重的社会负担,但却一直没有理想的治疗方案.继发性损伤是脊髓损伤后分子和细胞发生的一系列复杂的级联反应.它是SCI的重要组成部分,阐明继发性损伤的机制,针对不同的机制采取不同的干预措施是主要的治疗策略.而星形胶质细胞作为中枢神经系统中数量最多的细胞,在脊髓损伤后发挥了重要作用,受到研究人员的广泛关注,许多研究证明针刺可通过调节星形胶质细胞而发挥治疗作用,故本文综述了星形胶质细胞在脊髓损伤后的作用,以及针刺对其的干预影响,并提出日后研究的展望.