脉冲射频对坐骨神经分支损伤模型大鼠脊髓神经元凋亡的影响
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摘要
目的:探讨脉冲射频(pulsed radiofrequency,PRF)对坐骨神经分支损伤(spared nerve injury,SNI)模型大鼠脊髓神经元凋亡的影响。方法:将32只雄性SD大鼠随机分为4组(n=8):空白对照组(C组)、模型组(SNI组)、假治疗组(Sham组)和治疗组(PRF组)。于制模前0天,制模后1、4、7、10天,PRF治疗后6、12、24、36、48天分别测定4组大鼠机械缩足阈值(pain mechanical withdrawal threshold,PMWT);在PRF治疗前0天、治疗48天后分别处死每组大鼠4只,提取L4—L6节段脊髓,采用蛋白印记(Western Blot)技术检测bcl-2、caspase-3表达情况,TUNEL法检测脊髓组织凋亡细胞数。结果:(1)坐骨神经分支损伤术后,SNI组、Sham组和PRF组大鼠的PMWT较C组显著降低(P<0.01);(2)经PRF治疗后,PRF组大鼠的PMWT可显著提高(P<0.05);(3)随着PMWT提高,PRF组bcl-2蛋白表达量显著增加,caspase-3蛋白表达量显著降低(P<0.05),而脊髓组织细胞凋亡数量显著下降(P<0.05)。结论:PRF可能通过抑制脊髓神经细胞凋亡对SNI模型大鼠产生良好的镇痛作用。
Objective:To explore the effect of pulsed radiofrequency(PRF) for spinal cord neuron apoptosis in rat withspared nerve injury(SNI).Method:Thirty-two male SD rats were randomly divided into four groups,(1)control group(CON),(2)SNI model group(SNI),(3) sham treatment group(sham),(4) PRF treatment group(PRF). Pain mechanical withdrawal threshold(PMWT) was assessed before establishment of SNI model, 1,4,7 and 10 d after establishment of SNI model, and 6,12,24,36,48 d after PRF treatment. Four rats in each group were euthanized before PRF treatment and 48 days after treatment to collect L4—L6 spinal cord. Western blotting was conducted to assess the expression of bcl-2 and caspase-3 protein, and TUNEL method was used for apoptosis rate of spinal cord.Result:(1)The PMWT of SNI, Sham and PRF group were significantly decreased compared with CON group after spared nerve injury(P<0.01).(2)The PMWT of PRF group was obviously enhanced after PRF treatment(P<0.05).(3)The expression of bcl-2 was significantly up-regulated with the increase of PMWT while the expression of caspase-3 protein decreased(P<0.05), and the apoptosis rate was simultaneous decreased(P<0.05).Conclusion:PRF may produce good analgesic effect on SNI model rats by inhibiting the apoptosis of spinal cord neurons.
Objective:To explore the effect of pulsed radiofrequency(PRF) for spinal cord neuron apoptosis in rat withspared nerve injury(SNI).Method:Thirty-two male SD rats were randomly divided into four groups,(1)control group(CON),(2)SNI model group(SNI),(3) sham treatment group(sham),(4) PRF treatment group(PRF). Pain mechanical withdrawal threshold(PMWT) was assessed before establishment of SNI model, 1,4,7 and 10 d after establishment of SNI model, and 6,12,24,36,48 d after PRF treatment. Four rats in each group were euthanized before PRF treatment and 48 days after treatment to collect L4—L6 spinal cord. Western blotting was conducted to assess the expression of bcl-2 and caspase-3 protein, and TUNEL method was used for apoptosis rate of spinal cord.Result:(1)The PMWT of SNI, Sham and PRF group were significantly decreased compared with CON group after spared nerve injury(P<0.01).(2)The PMWT of PRF group was obviously enhanced after PRF treatment(P<0.05).(3)The expression of bcl-2 was significantly up-regulated with the increase of PMWT while the expression of caspase-3 protein decreased(P<0.05), and the apoptosis rate was simultaneous decreased(P<0.05).Conclusion:PRF may produce good analgesic effect on SNI model rats by inhibiting the apoptosis of spinal cord neurons.
引文
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[13]Chen NF, Chen WF, Sung CS, et al. Contributions of p38and ERK to the antinociceptive effects of TGF-beta1 in chronic constriction injury-induced neuropathic rats[J]. J Headache Pain,2016,17(1):72.
[14]Moore KA, Kohno T, Karchewski LA, et al. Partial peripheral nerve injury promotes a selective loss of GABAergic inhibition in the superficial dorsal horn of the spinal cord[J]. The Journal of Neuroscience,2002,22(15):6724—6731.
[15]Arons M, Pilmane M, Vasilevskis E, et al. Morphological changes in the lumbar dorsal root ganglion of the domestic porcine after pulsed radiofrequency stimulation[J]. Anesteziologiia I Reanimatologiia,2013,(4):26—30.
[16]Protasoni M, Reguzzoni M, Sangiorgi S, et al. Pulsed radiofrequency effects on the lumbar ganglion of the rat dorsal root:a morphological light and transmission electron microscopy study at acute stage[J].European Spine Journal,2009,18(4):473—478.
[17]Erdine S, Bilir A, Cosman ER, et al. Ultrastructural changes in axons following exposure to pulsed radiofrequency fields[J].Pain Practice,2009,9(6):407—417.
[18]Park CH, Lee YW, Kim YC, et al. Treatment experience of pulsed radiofrequency under ultrasound guided to the trapezius muscle at myofascial pain syndrome-a case report[J].The Korean Journal of Pain,2012,25(1):52—54.
[19]Chen KH, Yang CH, Juang SE, et al. Pulsed radiofrequency reduced complete Freund's adjuvant-induced mechanical hyperalgesia via the spinal c-Jun N-terminal kinase pathway[J].Cellular and Molecular Neurobiology,2014,34(2):195—203.
[2]吴飞翔,葛彦虎,缪雪蓉,等. N-乙酰半胱氨酸对大鼠神经元凋亡及神经病理性疼痛的影响[J].上海医学,2009,(6):494—497.
[3]吕时甲,窦智,蒋宗滨,等.脉冲射频对坐骨神经分支损伤模型大鼠机械痛阈的影响[J].实用疼痛学杂志,2014,10(5):329—333.
[4]李燕尧,张爱民,蒋宗滨,等.脉冲射频对保留性神经损伤大鼠Nav1.8 mRNA表达的影响[J].中国疼痛医学杂志,2015,12:903—907.
[5]Decosterd I, Woolf CJ. Spared nerve injury:an animal model of persistent peripheral neuropathic pain[J].Pain,2000,87(2):149—158.
[6]Lewis A, Hayashi T, Su TP, et al. Bcl-2 family in inter-organelle modulation of calcium signaling; roles in bioenergetics and cell survival[J]. Journal of Bioenergetics&Biomembranes, 2014, 46(1):1.
[7]Fujita T, Yoshimoto T, Matsuda S, et al. Interleukin-8 induces DNA synthesis, migration and down-regulation of cleaved caspase-3 in cultured human gingival epithelial cells[J]. Journal of Periodontal Research, 2015, 50(4):479—485.
[8]Chen SR, Pan HL. Effect of systemic and intrathecal gabapentin on allodynia in a new rat model of postherpetic neuralgia[J]. Brain Research,2005,1042(1):108—113.
[9]金小高,罗爱林,张广雄.三种大鼠神经病理性疼痛模型的制备和效果比较[J].临床麻醉学杂志,2005,05:338—340.
[10]Wu SX, Wang W, Li H, et al. The synaptic connectivity that underlies the noxious transmission and modulation within the superficial dorsal horn of the spinal cord[J]. Progress in Neurobiology,2010,91(1):38—54.
[11]Garrido-Suarez BB, Garrido G, Delgado R, et al. A Mangifera indica L. extract could be used to treat neuropathic pain and implication of mangiferin[J]. Molecules,2010,15(12):9035—9045.
[12]Basbaum AI, Bautista DM, Scherrer G, et al. Cellular and molecular mechanisms of pain[J]. Cell,2009,139(2):267—284.
[13]Chen NF, Chen WF, Sung CS, et al. Contributions of p38and ERK to the antinociceptive effects of TGF-beta1 in chronic constriction injury-induced neuropathic rats[J]. J Headache Pain,2016,17(1):72.
[14]Moore KA, Kohno T, Karchewski LA, et al. Partial peripheral nerve injury promotes a selective loss of GABAergic inhibition in the superficial dorsal horn of the spinal cord[J]. The Journal of Neuroscience,2002,22(15):6724—6731.
[15]Arons M, Pilmane M, Vasilevskis E, et al. Morphological changes in the lumbar dorsal root ganglion of the domestic porcine after pulsed radiofrequency stimulation[J]. Anesteziologiia I Reanimatologiia,2013,(4):26—30.
[16]Protasoni M, Reguzzoni M, Sangiorgi S, et al. Pulsed radiofrequency effects on the lumbar ganglion of the rat dorsal root:a morphological light and transmission electron microscopy study at acute stage[J].European Spine Journal,2009,18(4):473—478.
[17]Erdine S, Bilir A, Cosman ER, et al. Ultrastructural changes in axons following exposure to pulsed radiofrequency fields[J].Pain Practice,2009,9(6):407—417.
[18]Park CH, Lee YW, Kim YC, et al. Treatment experience of pulsed radiofrequency under ultrasound guided to the trapezius muscle at myofascial pain syndrome-a case report[J].The Korean Journal of Pain,2012,25(1):52—54.
[19]Chen KH, Yang CH, Juang SE, et al. Pulsed radiofrequency reduced complete Freund's adjuvant-induced mechanical hyperalgesia via the spinal c-Jun N-terminal kinase pathway[J].Cellular and Molecular Neurobiology,2014,34(2):195—203.
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