摘要
目的观察鞘内注射HPPE基因修饰的HEK293重组细胞(pLV-UbC-HPPE/HEK293),对骨癌痛大鼠行为学和脊髓背角pCREB表达的影响,探讨该重组细胞对大鼠骨癌痛的镇痛作用。方法雌性SD大鼠48只,随机分成两组,假手术(SHAM,12只)组和骨癌痛(CIBP,36只)组。大鼠左后肢胫骨打孔,SHA组大鼠注射生理盐水5ul,CIBP组大鼠注射MADB-106癌细胞5ul。于术前1d,术后7、14、21d分别测缩足反射潜伏期(PWL)X线下观察造模部位。将CIBP组随机分为CIBP′,CIBP′+GH,CIBP′+HH组,均12只。行鞘内穿刺并留置导管,分别注射PBS,GH和HH细胞悬液各20ul,隔天,共3次,测定PWL。CIBP′+HH组腹腔注射纳洛酮,再测PWL。实验结束后处死大鼠,取L4～L6脊髓,采用免疫组化法计数pCREB免疫反应阳性细胞数量。结果 (1)CIBP组术后7、14、21dPWL进行性下降(P<0.05),X线显示逐渐骨破坏;SHAM组差异无统计学意义(。2)CIBP′+HH组鞘内注射HH细胞后,PWL增高至(11.68±1.04)s,(P<0.05)(。3)CIBP′+HH组腹腔注射纳洛酮后PWL值降至(9.37±1.14)s,具有统计学意义(P<0.05),镇痛效应可被拮抗(。4)pCREB表达量:与CIBP′组相比,CIBP′+HH组降至(38.73±3.92),具有统计学意义(P<0.05)。结论鞘内注射HPPE基因修饰的HEK293细胞后,大鼠行为学变化和脊髓背角pCREB表达降低,表明该重组细胞对大鼠骨癌痛有镇痛效果。
Objective To observe the effect of intrathecal injection with HEK293 expressing human preproenkephalin gene steadily on the behavior and the expression of phosphorylated cAMP response element binding protein（pCREB） of the spinal dorsal in a ratmodel of bone cancer pain.Methods 48 female SD rats were randomly divided into 2 groups：SHAM（12,intra-tibial injection with normal saline in 5ul）.CIBP（36,intra-tibial injection with 5×106 MADB-106 mammary gland carcinoma cell in 5ul）.Changes of pain behavior and damages of the bone were asessed by paw-withdrawal latency and x-ray before injection as well as 7,14,21 day after injection.Then CIBP was randomly divided into 3 groups（n=12 each）：PBS,GH and HH cell at the dose of 20ul were administrated to rats in CIBP＇,CIBP＇＋GH and CIBP＇＋HH group respectively three times（at one day interval）.PWL assessment was finished.After intraperitoneal naloxone and another PWL assessment was finished,the rats were sacrificed by perfusion with 4% paraform.The lumbar 4-6 spinal cord was removed and sectioned on a freezing microtome,and then detected for the expression of pCREB through immunohistochemistry by image analysis method.Results（1）The PWL in CIBP group was significantly decreased on the 7,14,21 days after operation.But SHAM group remained unchanged.The x-ray of the tibia showed clear bone destruction.（2） After intrathecal injection,the PWL in CIBP＇＋HH group was increased to（11.68±1.04）s（P〈0.05）.（3） The PWL in CIBP＇＋HH group was decreased to（9.37±1.14）s,and it＇s efficacy was reversed by intraperitoneal naloxone.（4） Compared with the CIBP＇group,the expression of pCREB of CIBP＇＋HH group was decreased to（38.73±3.92）（P〈0.05）.Conclusion Intrathecal injection with the cell of plv-UbC-HPPE/HEK293 had significant analgesic effects on rats with bone cancer pain via pain behavior and the expression of pCREB of the spinal dorsal.
出处
《当代医学》
2012年第17期1-4,共4页
Contemporary Medicine
基金
山西省科技攻关项目(20090311058-2)
