鞘内注射shRNA慢病毒载体缓解慢性炎性疼痛的机制研究  

Mechanism of intrathecal injection of shRNA lentivirus vector in relieving chronic inflammatory pain

作　　者：姜晓阳[1] 冯艳茹[2] 刘亚华[1] 侯彦深[1] JIANG Xiao-yang;FENG Yan-ru;LIU Ya-hua;HOU Yan-shen(Anesthesia and Perioperative Medicine Center,Cancer Hospital Affiliated to Xinjiang Medical University,Urumqi Xinjiang 830011,China;Department of Nutrition,Cancer Hospital Affiliated to Xinjiang Medical University,Urumqi Xinjiang 830011,China)

机构地区：[1]新疆医科大学附属肿瘤医院麻醉与围术期医学中心,新疆乌鲁木齐830011 [2]新疆医科大学附属肿瘤医院营养科,新疆乌鲁木齐830011

出　　处：《局解手术学杂志》2023年第6期494-498,共5页Journal of Regional Anatomy and Operative Surgery

基　　金：新疆维吾尔自治区自然科学基金(2017D01C372)。

摘　　要：目的探究鞘内注射shRNA慢病毒载体对慢性炎性疼痛大鼠疼痛行为学及脊髓胶质纤维酸性蛋白(GFAP)、OX42蛋白表达的影响。方法将54只SD大鼠随机分为正常组、空病毒组、慢病毒载体组,采用足底注射完全弗氏佐剂(CFA)建立大鼠慢性炎性疼痛模型,正常组大鼠足底注射生理盐水,鞘内注射PBS缓冲液;空病毒组和慢病毒载体组大鼠足底注射CFA,鞘内分别注射空病毒和慢病毒液。于建模前1 d及建模后第1、3、6、9、13天测量机械缩足阈值(PMWT)及热缩足潜伏期(PTWL);于建模后第3、6、13天留取大鼠L4~L5背根神经节(DRG)组织和脊髓节段组织,检测Nav1.7和GFAP、OX42蛋白表达水平。结果与正常组大鼠比较,空病毒组及慢病毒载体组大鼠PMWT值以及慢病毒载体组大鼠PTWL值在建模后第1天明显下降,在建模后第3天下降达最低值,之后逐渐升高;空病毒组PTWL值在建模后第1天明显下降,在建模后第3天下降达最低值,之后较第3天升高,但仍低于基础值。与空病毒组大鼠比较,慢病毒载体组大鼠PMWT、PTWL值从建模后第3天开始逐渐升高,差异有统计学意义(P<0.05)。在建模后第3、6、13天,正常组大鼠DRG中Nav1.7及脊髓GFAP、OX42蛋白表达水平基本无变化;空病毒组和慢病毒载体组大鼠DRG中Nav1.7及脊髓GFAP、OX42蛋白表达水平较正常组增加,差异有统计学意义(P<0.05);慢病毒载体组大鼠DRG中Nav1.7及脊髓GFAP、OX42蛋白表达水平较空病毒组下降,差异有统计学意义(P<0.05)。结论鞘内注射shRNA慢病毒载体可抑制大鼠DRG中Nav1.7表达,抑制脊髓胶质细胞活化,下调GFAP、OX42蛋白表达,缓解大鼠慢性炎性疼痛。Objective To investigate the effects of intrathecal injection of shRNA lentivirus vector on pain behavior and expression of spinal cord glial fibrillary acidic protein(GFAP)and OX42 protein in rats with chronic inflammatory pain.Methods A total of 54 SD rats were randomly divided into the normal group,the empty virus group and the lentivirus vector group.The rat model of chronic inflammatory pain was established by plantar injection of complete Freund's adjuvant(CFA).Rats in the normal group were injected with normal saline at the sole and PBS buffered solution in the sheath,while rats in the empty virus group and the lentivirus vector group were injected with CFA at the sole,and the empty virus and lentivirus solutions were injected into the sheaths,respectively.The paw mechanical withdrawal threshold(PMWT)and paw thermal withdrawal latency(PTWL)were measured 1 day before modeling and 1,3,6,9 and 13 days after modeling.The L4 to L5 dorsal root ganglia(DRG)tissues and spinal cord segments tissues of rats were collected 3,6 and 13 days after modeling,and the expression levels of Nav1.7,GFAP and OX42 proteins were measured.Results Compared with the normal group,the values of PMWT in the empty virus group and the lentivirus vector group and the value of PTWL in the lentivirus vector group decreased significantly on the first day after modeling,and reached the lowest value on the third day after modeling,and then increased gradually;the value of PTWL in the empty virus group decreased significantly on the first day after modeling,and reached the lowest value on the third day after modeling,then increased gradually,but it was still lower than the basic value.Compared with the empty virus group,the values of PMWT and PTWL in the lentivirus vector group gradually increased from the third day after modeling,and the difference was statistically significant(P<0.05).On the 3rd,6th and 13th day after modeling,there was no significant difference in the expression levels of Nav1.7 in DRG,GFAP and OX42 proteins in spinal cord o

关 键 词：鞘内注射 慢病毒载体 慢性炎性 疼痛行为 

分 类 号：R363[医药卫生—病理学]