CAJ | 학술논문

目的评价皮下转染重组腺病毒介导的人神经生长因子β(Ad-hNGFβ)基因对糖尿病神经病理性痛大鼠感觉神经功能的影响.方法健康雄性SD大鼠,体重180 ～ 220 g,采用腹腔注射链脲霉素(STZ)75 mg/kg的方法制备大鼠糖尿病模型.取10只大鼠作为对照组(C组),不制备糖尿病模型；取75只糖尿病模型制备成功的大鼠,采用随机数字表法,将其分为3组(n=25):糖尿病神经病理性痛组(DNP组)、Ad-hNGFβ基因治疗组(Ad-NGF组)和LacZ重组复制缺陷性腺病毒基因治疗组(Ad-LacZ组).Ad-NGF组和Ad-LacZ组分别于STZ注射后21 d痛阈测定结束后双侧腹股沟皮下脂肪接种1.12× 1011PFU Ad-hNGFβ 10ul和1.12× 1011PFU Ad-LacZ 10ul.分别于STZ注射前(基础状态)、注射后21、35和49 d时测定机械痛阈和热痛阈.于STZ注射后49 d痛阈测定结束后,测定感觉神经传导速度和潜伏期,采用免疫组化法测定背根神经节P物质的表达.结果与C组比较,DNP组、Ad-NGF组和Ad-LacZ组STZ注射后各时点机械痛阈和热痛阈降低,DNP组和Ad-LacZ组感觉神经传导速度减慢,潜伏期延长,背根神经节P物质表达下调,Ad-NGF组感觉神经传导速度减慢(P＜0.05),潜伏期和背根神经节P物质表达差异无统计学意义(P＞0.05).与DNP组比较,Ad-NGF组STZ注射后49d时热痛阈升高,感觉神经传导速度加快,潜伏期缩短,背根神经节P物质表达上调(P＜ 0.05或0.01),机械痛阈差异无统计学意义,Ad-LacZ组机械痛阈、热痛阈、感觉神经传导速度、潜伏期和背根神经节P物质表达差异无统计学意义(P＞0.05).结论 Ad-hNGFβ基因转染可改善糖尿病神经病理性痛大鼠的感觉神经功能,其机制可能与上调背根神经节P物质的表达有关.
목적 평개피하전염중조선병독개도적인신경생장인자β(Ad-hNGFβ)기인대당뇨병신경병이성통대서감각신경공능적영향.방법 건강웅성SD대서,체중180 ～ 220 g,채용복강주사련뇨매소(STZ)75 mg/kg적방법제비대서당뇨병모형.취10지대서작위대조조(C조),불제비당뇨병모형；취75지당뇨병모형제비성공적대서,채용수궤수자표법,장기분위3조(n=25):당뇨병신경병이성통조(DNP조)、Ad-hNGFβ기인치료조(Ad-NGF조)화LacZ중조복제결함성선병독기인치료조(Ad-LacZ조).Ad-NGF조화Ad-LacZ조분별우STZ주사후21 d통역측정결속후쌍측복고구피하지방접충1.12× 1011PFU Ad-hNGFβ 10ul화1.12× 1011PFU Ad-LacZ 10ul.분별우STZ주사전(기출상태)、주사후21、35화49 d시측정궤계통역화열통역.우STZ주사후49 d통역측정결속후,측정감각신경전도속도화잠복기,채용면역조화법측정배근신경절P물질적표체.결과 여C조비교,DNP조、Ad-NGF조화Ad-LacZ조STZ주사후각시점궤계통역화열통역강저,DNP조화Ad-LacZ조감각신경전도속도감만,잠복기연장,배근신경절P물질표체하조,Ad-NGF조감각신경전도속도감만(P＜0.05),잠복기화배근신경절P물질표체차이무통계학의의(P＞0.05).여DNP조비교,Ad-NGF조STZ주사후49d시열통역승고,감각신경전도속도가쾌,잠복기축단,배근신경절P물질표체상조(P＜ 0.05혹0.01),궤계통역차이무통계학의의,Ad-LacZ조궤계통역、열통역、감각신경전도속도、잠복기화배근신경절P물질표체차이무통계학의의(P＞0.05).결론 Ad-hNGFβ기인전염가개선당뇨병신경병이성통대서적감각신경공능,기궤제가능여상조배근신경절P물질적표체유관.
Objective To evalute the effects of subcutaneous transfection of human beta-nerve growth factor (Ad-hNGFβ) gene on the function of sensory nerve in a rat model of diabetic neuropathic pain (DNP).Methods Healthy male Sprague-Dawley rats,weighing 180-220 g,were used in this study.Ten rats were randomly collected as normal control without DNP (group C).DNP model was established by intraperitoneal injection of streptozocin (STZ) 75 mg/kg.Seventy-five rats with DNP were randomly divided into 3 groups (n =25 each):DNP group; Ad-hNGF group and Ad-LacZ group.Groups Ad-NGF and Ad-LacZ received subcutaneous transfection of 1.12 × 1011 PFU Ad-hNGFβ 10 ul and 1.12 × 1011 PFU Ad-LacZ 10 ul respectively after pain thresholds were measured on 21 days after STZ injection.The mechanical and thermal pain thresholds were measured before STZ injection (baseline) and at 21,35 and 49 days after STZ injection.The conduction velocity and latency of the sensory nerve and the expression of substance P (SP) in dorsal root ganglion (by immmuno-histpchemistry) were determined after measurement of pain threshold on 49 days after STZ injection.Results Compared with group C,the mechanical and thermal pain thresholds were significantly increased at each time point after STZ injection in groups DNP,Ad-NGF and Ad-Lacz,the conduction velocity of the sensory nerve was significantly decreased,the latency of the sensory nerve was prolonged,and the expression of SP in dorsal root ganglion was down-regulated in groups DNP and Ad-LacZ,the conduction velocity of the sensory nerve was significantly decreased (P ＜ 0.05),and no significant change was found in the latency of the sensory nerve and expression of SP in dorsal root ganglion in group Ad-NGF (P ＞ 0.05).Compared with group DNP,the thermal pain threshold was significantly increased at 49 days after STZ injection,the conduction velocity of the sensory nerve was decreased,the latency of the sensory nerve was shortened,and the expression of SP in dorsal root ganglion was down-regulated (P ＜ 0.05 or 0.01),while no significant change was found in the mechanical pain threshold in group Ad-NGF,and no significant change was found in the parameters mentioned above in group Ad-LacZ (P ＞ 0.05).Conclusion Subcutaneous transfection of Ad-hNGFβ can improve the function of sensory nerve in a rat model of DNP through up-regulat-ing of the SP expression in dorsal root ganglion.