CAJ | 학술논문

目的评价脊髓背角神经元磷脂酰肌醇-3激酶(PI3K) p110β在大鼠关节炎性痛形成中的作用及与辣椒素受体(TRPV1)及酸敏感离子通道1a(ASIC1a)的关系.方法鞘内置管成功的成年雌性SD大鼠40只,3月龄,体重250 ～ 300 g,采用随机数字表法,将其分为4组(n=10)∶对照组(C组)、关节炎性痛组(AP组)、AP+ PI3K p110β错义寡核苷酸组(MS组)和AP+ PI3K p110β反义寡核苷酸组(AS组).采用右踝关节腔内注射完全弗氏佐剂建立关节炎性痛模型,模型制备后即刻AP组、MS组、AS组分别经鞘内注射生理盐水、错义寡核苷酸15 μg和反义寡核苷酸15 μg,容量20μl,1次/d,连续6d.于术前1d、术后4、7、10d时测定机械缩足反应阈(MWT)和热缩足反应潜伏期(TWL),术后10 d处死大鼠,取腰段脊髓,采用Western blot法检测脊髓背角神经元PI3K p110β表达,采用免疫组化法检测脊髓背角神经元TRPV1和ASICla的表达.结果与C组比较,AP组、MS组和AS组术后各时点MWT降低,TWL缩短,脊髓背角神经元PI3K p110β、TRPV1和ASICla表达上调(P＜0.01)；与AP组和MS组比较,AS组术后各时点MWT升高,TWL延长,脊髓背角神经元PI3K p110β、TRPV1和ASIC1a表达下调(P＜0.01).结论脊髓背角神经元PI3K p110β参与大鼠关节炎性痛的形成,其机制与上调脊髓背角神经元TRPVl和ASICla的表达有关.
목적 평개척수배각신경원린지선기순-3격매(PI3K) p110β재대서관절염성통형성중적작용급여랄초소수체(TRPV1)급산민감리자통도1a(ASIC1a)적관계.방법 초내치관성공적성년자성SD대서40지,3월령,체중250 ～ 300 g,채용수궤수자표법,장기분위4조(n=10)∶대조조(C조)、관절염성통조(AP조)、AP+ PI3K p110β착의과핵감산조(MS조)화AP+ PI3K p110β반의과핵감산조(AS조).채용우과관절강내주사완전불씨좌제건립관절염성통모형,모형제비후즉각AP조、MS조、AS조분별경초내주사생리염수、착의과핵감산15 μg화반의과핵감산15 μg,용량20μl,1차/d,련속6d.우술전1d、술후4、7、10d시측정궤계축족반응역(MWT)화열축족반응잠복기(TWL),술후10 d처사대서,취요단척수,채용Western blot법검측척수배각신경원PI3K p110β표체,채용면역조화법검측척수배각신경원TRPV1화ASICla적표체.결과 여C조비교,AP조、MS조화AS조술후각시점MWT강저,TWL축단,척수배각신경원PI3K p110β、TRPV1화ASICla표체상조(P＜0.01)；여AP조화MS조비교,AS조술후각시점MWT승고,TWL연장,척수배각신경원PI3K p110β、TRPV1화ASIC1a표체하조(P＜0.01).결론 척수배각신경원PI3K p110β삼여대서관절염성통적형성,기궤제여상조척수배각신경원TRPVl화ASICla적표체유관.
Objective To evaluate the role of phosphatidylinositol 3-kinase (PI3K) p110β in spinal dorsal horn neurons in the development of arthritic pain (AP) in rats and the relationship with transient receptor potential vanilloid 1 (TRPV1) and acid-sensing ion channel (ASIC)1 a.Methods Forty adult female Sprague-Dawley rats in which intrathecal catheters were successfully placed,aged 3 months,weighing 250-300 g,were randomly divided into 4 groups (n =10 each):control group (group C),group AP,AP + PI3K p110β missense oligo-deoxynucleotide group (group MS) and AP + PI3K p110β antisense oligo-deoxynucleotide group (group AS).AP was induced by injecting complete Freund's adjuvant into the ankle joint cavity of right hindpaw.Normal saline 20 μl,missense oligo-deoxynucleotide 15 μg (20μl) and antisense oligo-deoxynucleotide 15 μg (20 μl) were administered intrathecally once a day for 6 consecutive days starting from the time immediately after arthritis was induced in groups AP,MS and AS,respectively.Mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured 1 day before operation (T0) and on days 4,7,10 after operation (T1-3).The rats were then sacrificed after the last measurement of pain threshold at T3.L4-6 segment of the spinal cord was removed for detection of expression of PI3K p110β (by Western blot),and TRPV1 and ASICla (by immunohistochemistry)in spinal dorsal horn neurons.Results Compared with group C,MWT and TWL were significantly decreased atT1-3,and the expression of PI3K p110β,TRPV1 and ASIC1a was up-regulated in the other 3 groups(P＜ 0.01).MWT and TWL were significantly higher at T1-3,and the expression of PI3K p110β,TRPV1 and ASIC1a was lower in group AS than in groups AP and MS (P ＜ 0.01).Conclusion PI3K p110β in spinal dorsal horn neurons is involved in the development of AP in rats,and the mechanism is related to up-regulation of TRPV1 and ASIC1a expression in spinal dorsal horn neurons.