摘要
目的构建受强力霉素诱导表达脑啡肽的永生化大鼠星形胶质细胞株,并评价鞘内移植该细胞对慢性坐骨神经压榨性损伤(CCI)大鼠的镇痛效应。方法应用逆转录病毒转染法,建立受强力霉素调控表达人前脑啡肽原基因(hPPE)的永生化大鼠星形胶质细胞株(IAST/Tet-On/hPPE),实时定量PCR及放射免疫分析法检测强力霉素对该细胞株脑啡肽表达的定量调节。将IAST/Tet-On/hPPE细胞植入CCI大鼠蛛网膜下腔,观察腹腔注射强力霉素对其镇痛效应的影响及免疫组织化学检测脊髓背角Fos蛋白的表达。结果实时定量PCR及放射免疫分析结果显示,强力霉素可定量调控IAST/Tet-On/hPPE细胞株中脑啡肽的表达;IAST/Tet-On/hPPE植入CCI大鼠的蛛网膜下腔后,大鼠机械缩爪阈值升高(P<0.05),脊髓背角浅层Fos蛋白表达明显降低(P<0.05),且该镇痛效应受强力霉素调控。结论成功构建了可调控的定量表达脑啡肽的永生化星形胶质细胞株,其有望成为慢性疼痛治疗的新方法。
Objective To establish an immortalized rat astrocyte strain （IAST） expressing enkephalin regulated by doxycycline （Dox） and observe its analysesic effect on rat chronic neuropathic pain. Methods Retrovirus infection method was employed to develop an immortalized rat astrocyte strain expression enkephalin regulated by doxycycline, hPPE gene expression level of IAST/Tet-On/hPPE strain was detected by Real time-PCR and radioimmunoassay. Its analgesic potential was investigated by mechanical paw withdrawal thresholds after these cells were implanted into the subarachnoid space of chronic constrictive injury （CCI） rats. The expression of Fos protein in the dorsal horn of spinal cord was determined by immunohistochemistry. Results An immortalized rat astrocyte strain secreting enkephalin under the control of doxycycline was established successfully. After transplantation of IAST/Tet-On/hPPE cell into the subarachnoid space of chronic constrictive injury （CCI） rats, the sensitivity of mechanical allodynia and the expression of Fos protein were significantly decreased （ P 〈 0.05）, so the transplantation of lAST/ Tet-On/hPPE cell alleviated significantly CCI-induced chronic neuropathic pain in rats and the analgesic effect was also able to be regulated by Dox. Conclusion An immortalized rat astrocyte strain expressing enkephalin regulated by Dox has been established, which may provide a new tool for regulatable gene therapy for chronic pain in the future.
出处
《解剖学报》
CAS
CSCD
北大核心
2006年第6期617-621,共5页
Acta Anatomica Sinica
基金
国家自然科学基金资助项目(30170905)
杨森基金资助
