论文部分内容阅读
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 construct an immortalized rat astrocyte cell line induced by doxycycline and to evaluate the analgesic effect of intrathecal transplantation of the cells on chronic constriction injury (CCI) in rats. Methods The retroviral transfection method was used to establish the immortalized rat astrocyte line (IAST / Tet-On / hPPE) regulated by doxycycline (hPPE) Radioimmunoassay was used to detect the quantitative regulation of enkephalin expression in this cell line. IAST / Tet-On / hPPE cells were implanted into the subarachnoid space of CCI rats to observe the effect of intraperitoneal injection of doxycycline on the analgesic effect and the expression of Fos protein in spinal dorsal horn by immunohistochemistry. Results Real-time quantitative PCR and radioimmunoassay results showed that doxycycline could quantitatively regulate the expression of enkephalin in IAST / Tet-On / hPPE cell lines. IAST / Tet-On / hPPE was implanted into the subarachnoid space of CCI rats (P <0.05). The expression of Fos protein in the spinal dorsal horn was significantly decreased (P <0.05), and the analgesic effect was controlled by doxycycline. Conclusion An immortalized astrocyte cell line that can regulate the expression of enkephalin was successfully constructed, which is expected to become a new method for the treatment of chronic pain.