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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 investigate the effect of intrathecal injection of HPPE gene-modified HEK293 recombinant cells (pLV-UbC-HPPE/HEK293) on the behavior of bone cancer pain rats and the expression of pCREB in dorsal horn of spinal cord, and to explore the effects of the recombinant cells on rat bone cancer pain. Analgesic effect. Methods Forty-eight female SD rats were randomly divided into two groups: sham-operated (SHAM, 12) and bone cancer pain (CIBP, 36) groups. The left hind limb of the rat was perforated, SHA rats were injected with 5 ul normal saline, and CIBP group rats were injected with 5 μl of MADB-106 cancer cells. At 1 day before surgery and 7, 14 and 21 days after surgery, the PWL was observed under the X-ray to observe the model site. The CIBP group was randomly divided into CIBP′, CIBP′+GH, and CIBP′+HH groups, with 12 rats in each group. Underwent intrathecal puncture and indwelling catheter, respectively injected PBS, GH and HH cell suspension 20ul each, the next day, a total of 3 times, determination of PWL. In the CIBP’+HH group, naloxone was injected intraperitoneally and PWL was measured. After the end of the experiment, the rats were sacrificed, L4 to L6 spinal cords were taken and the number of pCREB immunoreactive cells was counted by immunohistochemistry. Results (1) PWL was progressively decreased at 7, 14, and 21 d in CIBP group (P<0.05). X-ray showed gradual bone destruction. There was no significant difference in SHAM group.(2) CIBP′+HH group intrathecal HH cells Afterwards, the PWL increased to (11.68±1.04)s, (P<0.05).(3) The PWL value after intraperitoneal injection of naloxone in the CIBP′+HH group decreased to (9.37±1.14) s, which was statistically significant (P< 0.05), the analgesic effect can be antagonized (. 4) pCREB expression: Compared with CIBP′ group, CIBP′+HH group decreased to (38.73±3.92), with statistical significance (P<0.05). Conclusion The changes of behavior and the expression of pCREB in the dorsal horn of spinal cord were reduced after intrathecal injection of HPPE gene-modified HEK293 cells, indicating that the recombinant cells have analgesic effect on rat bone cancer pain.