愈痫灵方对难治性癫痫大鼠多药耐药相关蛋白MRP1、P-gp、LAP、MCP-1表达的作用

目的：探讨“愈痫灵”方对红藻氨酸（KA）致痫难治性癫痫模型大鼠海马及颞叶皮质区多药耐药相关蛋白1（MRP1）、P糖蛋白（P-gp）、层黏连蛋白（LAP）以及单核细胞趋化蛋白1(MCP-1)表达的影响。方法（1）造模：在脑立体定位仪引导下，海马区微量进样器注入KA1.0μL（即1.0μg）点燃发作，选取发作行为级别在Racines标准Ⅳ级以上者，以丙戊酸钠及卡马西平灌胃干预14 d后，再次以亚惊厥剂量KA0.5μL复燃，筛选再次发作级别在Ⅳ级以上或持续状态大鼠，且脑电图检测有癫痫样波放电者为造模成功的耐药难治性癫痫模型鼠。（2）分组与处理：造模成功鼠随机分为愈痫灵方干预组、拉莫三嗪对照组、模型组，另设假手术对照组、空白对照组共5组，分别给予“愈痫灵”汤剂、拉莫三嗪及同等体积的蒸馏水(2 mL/d)灌胃30 d。（3）标本处置与检测：采用免疫组织化学技术，检测多药耐药相关蛋白MRP1、P-gp、LAP、MCP-1在海马与颞叶皮层的表达。结果与假手术对照组、空白对照组间比较，造模成功组海马区及颞叶皮质区MRP1、P-gp、LAP、MCP-1的表达均明显升高，差异有统计学意义(P<0.01)；与模型组比较，愈痫灵、拉莫三嗪干预处理后能显著降低模型鼠海马区MRP1、P-gp表达水平（P<0.05），海马区及颞叶皮质区LAP、MCP-1的表达差异均无统计学意义(P>0.05)；愈痫灵方组与拉莫三嗪组比较，差异均无统计学意义(P>0.05)；各组间颞叶皮质区LAP、MCP-1表达差异无统计学意义(P>0.05)。结论耐药性癫痫大鼠模型海马及颞叶皮质区多药耐药相关蛋白MRP1、P-gp、LAP、MCP-1表达均明显升高，逆转与降低海马区MRP1、P-gp的表达可能是“愈痫灵”方抗耐药性癫痫作用机制之一。

Effects of YuXianLing Decoction on the Expressions of Multiple Drug Resistant Related Proteins MRP1, P-gp,LAP MCP-1 in Refractory Epileptic Rats Induced by KA

〔Abstract〕 Objective To investigate the effects of YuXianLing Decoction (YXLD) on the expressions of multi-drug resistance related protein 1 (MRP1), P-glycoprotein(P-gp), layer ankylosis protein(LAP) and Monocyte chemo attractant protein-1 (MCP-1) in hippocampus and temporal lobe cortex of the refractory epileptic rats induced by kaicic acid (KA). Methods (1) Rats models: Hippocampus located by brain stereotactic apparatus according to Paxions Watras map was micro injected 1.0 μL KA to kindle seizures. The rats with above seizure behavior surpass Ⅳ level were intragastric administration interfered by sodium valproate and carbamazepine for 14 days and rekindled with 0.5 μL KA. The rats with above seizure behavior surpass Ⅳ level and epileptiform discharge waves were selected to be successful resistant refractory epilepsy model rats. (2) Grouping and handling: The model rats were randomLy divided into YXLD group, Lamotrigine (LTG)control group and model group, and the sham-operation group and blank control group were also setted. The rats of all groups were intragastric administration interfered by the same volume gavage (2 mL/d) of distilled water, LTG and YXLD for 30 days, respectively. (3) Disposition and detection of specimen: the expressions of MRP1, P-gp, LAP, MCP-1 were detected by immunohistochemical technique. Results Compared with the blank control group and sham-operation group, the expression of successful model rats of MRP1, P-gp, LAP and MCP-1 in hippocampus and temporal cortex were significantly increased,the difference was statistically significant (P<0.01). Compared with the model group, YXLD and LTG intervention can significantly reduce the expression level of MRP1 and P-gp in hippocampus area (P<0.05). The expression of LAP and MCP-1 in hippocampus and temporal lobe cortex were not statistically significant (P>0.05). There were no significant differeces between YXLD group and LAP group (P>0.05). The expressions of LAP, MCP-1 in temporal cortex among groups showed no significant difference (P>0.05). Conclusion The expression of MRP1, P-gp, LAP, MCP-1 of multiply drug resistance in hippocampus and temporal lobe cortex areas of model rats were significantly increased. Reverse and decrease the expression of MRP1 and P-gp in the hippocampal area may be one of the mechanisms of anti-drug resistant epilepsy of YXLD.