CAJ | 학술논문

目的：评价鞘内注射TRESK基因重组腺病毒对神经病理性痛大鼠脊髓趋化因子介导炎性反应的影响。方法健康雄性SD大鼠36只，体重200～250 g，采用随机数字表法分为6组（ n＝6）：对照组（ C组）、假手术组（ S 组）、神经病理性痛组（ NP 组）、TRESK 过表达腺病毒组（ TRESK组）、阴性腺病毒组（ Virus组）和生理盐水组（ NS组）。采用坐骨神经分支选择性损伤法制备大鼠神经病理性痛模型。 TRESK组、NS组和Virus组于造模成功后即刻分别鞘内注射pAd∕CMV∕V5?DEST?TRESK 25μl（109IU∕ml）、阴性腺病毒25μl和生理盐水25μl。于造模前1 d（T0）和造模后1、3、7和14 d（ T1～4）时测定机械缩足反应阈（ MWT）和热缩足潜伏期（ TWL）。于T3时测定痛阈后处死大鼠取脊髓，采用PCR法检测单核细胞趋化因子?1（ MCP?1）、巨噬细胞炎性蛋白?2（ MIP?2）、TNF?α、IL?1β和IL?6的mRNA的表达。结果各组不同时点TWL比较差异无统计学意义（ P＞0．05）。与C组和S组比较，NP组和TRESK组T1～4时、Virus组和NS组T1～3时MWT降低，NP 组、TRESK组、Virus组和NS组脊髓MCP?1、MIP?2、IL?1β、IL?6和TNF?α的mRNA表达上调（P＜0．05）；与NP 组比较，TRESK组T1?4时MWT升高，脊髓MCP?1、MIP?2、IL?1β、IL?6和TNF?α的mRNA表达下调（P＜0．05）。结论鞘内注射TRESK基因重组腺病毒减轻大鼠神经病理性痛的机制与抑制脊髓趋化因子介导的炎性反应有关。
목적：평개초내주사TRESK기인중조선병독대신경병이성통대서척수추화인자개도염성반응적영향。방법건강웅성SD대서36지，체중200～250 g，채용수궤수자표법분위6조（ n＝6）：대조조（ C조）、가수술조（ S 조）、신경병이성통조（ NP 조）、TRESK 과표체선병독조（ TRESK조）、음성선병독조（ Virus조）화생리염수조（ NS조）。채용좌골신경분지선택성손상법제비대서신경병이성통모형。 TRESK조、NS조화Virus조우조모성공후즉각분별초내주사pAd∕CMV∕V5?DEST?TRESK 25μl（109IU∕ml）、음성선병독25μl화생리염수25μl。우조모전1 d（T0）화조모후1、3、7화14 d（ T1～4）시측정궤계축족반응역（ MWT）화열축족잠복기（ TWL）。우T3시측정통역후처사대서취척수，채용PCR법검측단핵세포추화인자?1（ MCP?1）、거서세포염성단백?2（ MIP?2）、TNF?α、IL?1β화IL?6적mRNA적표체。결과각조불동시점TWL비교차이무통계학의의（ P＞0．05）。여C조화S조비교，NP조화TRESK조T1～4시、Virus조화NS조T1～3시MWT강저，NP 조、TRESK조、Virus조화NS조척수MCP?1、MIP?2、IL?1β、IL?6화TNF?α적mRNA표체상조（P＜0．05）；여NP 조비교，TRESK조T1?4시MWT승고，척수MCP?1、MIP?2、IL?1β、IL?6화TNF?α적mRNA표체하조（P＜0．05）。결론초내주사TRESK기인중조선병독감경대서신경병이성통적궤제여억제척수추화인자개도적염성반응유관。
Objective To evaluate the effects of intrathecal TRESK gene recombinant adenovirus on inflammatory responses mediated by chemokine in the spinal cord of rats with neuropathic pain ( NP ) . Methods Thirty?six male Sprague?Dawley rats, weighing 200-250 g, were randomly divided into 6 groups (n=6 each) using a random number table: control group (group C); sham operation group (group S);NP group; TRESK?overexpressed adenovirus group ( group TRESK ); negative adenovirus group ( group Virus); normal saline group ( group NS) . Spinal nerve injury was produced by exposing the sciatic nerve and its branches and ligation and transection of tibial nerve and common fibular nerve in anesthetized rats. In TRESK, Virus and NS groups, pAd∕CMV∕V5?DEST?TRESK 25 μl (109IU∕ml), negative adenovirus 25 μl and normal saline 25 μl were intrathecally injected, respectively. At 1 day before operation ( base?line, T0 ) and 1, 3, 7 and 14 days after operation ( T1-4 ) , the mechanical paw withdrawal threshold ( MWT) and thermal paw withdrawal latency were measured. Six rats in each group were sacrificed after measurement of pain threshold at T3 . The L4,5 segments of the spinal cords were removed for determination of monocyte chemotactic protein?1 ( MCP?1) , MIP?2, tumor necrosis factor?alpha ( TNF?α) , interleukin?1 beta ( IL?1β) and IL?6 mRNA expression by real?time PCR. Results There was no significant difference in thermal paw withdrawal latency at each time point between groups. Compared with C and S groups, MWT at T1-4 in NP and TRESK groups and at T1-3 in Virus and NS groups were significantly decreased, and the expression of MCP?1, MIP?2, TNF?α, IL?1βand IL?6 mRNA was up?regulated in NP, TRESK, Virus and NS groups. Compared with group NP, MWT was significantly increased at T1-4, and the expres?sion of MCP?1, MIP?2, TNF?α, IL?1β and IL?6 mRNA was down?regulated in group TRESK. Conclusion The mechanism by which intrathecal TRESK gene recombinant adenovirus reduces NP is re?lated to inhibition of inflammatory responses mediated by chemokine in the spinal cord of rats.