湖南中医药大学学报
湖南中醫藥大學學報
호남중의약대학학보
Journal of Traditional Chinese Medicine University of Hunan
2015年
11期
18-21
,共4页
李振光%宋祖丽%王净净%李智雄%谢静涛%左亚杰%张曦%肖瑶
李振光%宋祖麗%王淨淨%李智雄%謝靜濤%左亞傑%張晞%肖瑤
리진광%송조려%왕정정%리지웅%사정도%좌아걸%장희%초요
耐药性癫痫%愈痫灵方%多药耐药基因MDR1b%红藻氨酸
耐藥性癲癇%愈癇靈方%多藥耐藥基因MDR1b%紅藻氨痠
내약성전간%유간령방%다약내약기인MDR1b%홍조안산
drug resistant epilepsy%Yuxianling Decoction%multidrug resistance gene MDR1b%kainic acid
目的:探讨愈痫灵方对耐药性癫痫模型大鼠海马及颞叶皮质多药耐药基因MDR1b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红藻氨酸(KA)1.0μL(浓度为1.0μg/μL)点燃造模,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,再次亚惊厥剂量0.5μL KA复燃,选择再次发作级别在Ⅳ级以上或持续状态大鼠,且脑电图检测有癫痫样波发放者筛选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、空白对照组共计5组,每组12只。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:采用荧光实时定量聚合酶链反应(RT-PCR)的方法,分别检测海马区与颞叶皮层MDR1b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MDR1b的表达水平明显升高(P<0.01),颞叶皮质区差异无统计学意义(P>0.05);所有造模组的海马区MDR1b表达高于颞叶皮质,差异有统计学意义(P<0.01)。与模型组比较,愈痫灵、拉莫三嗪干预后能显著降低KA致痫鼠海马区MDR1b的表达水平(P<0.05);愈痫灵方组与拉莫三嗪组间比较,海马与颞叶皮质之间差异均无统计学意义(P>0.05);各组间颞叶皮质区MDR1b表达差异无统计学意义(P>0.05)。结论 KA造模耐药性癫痫大鼠模型海马区多药耐药基因MDR1b的表达较皮质区明显升高。愈痫灵方降低海马区MDR1b的表达,可能是其抗耐药性癫痫作用机制之一。
目的:探討愈癇靈方對耐藥性癲癇模型大鼠海馬及顳葉皮質多藥耐藥基因MDR1b錶達的影響。方法①造模:在腦立體定位儀引導下,海馬區微量進樣器註入紅藻氨痠(KA)1.0μL(濃度為1.0μg/μL)點燃造模,選取髮作行為級彆在Racines標準Ⅳ級以上者,以丙戊痠鈉及卡馬西平灌胃榦預14 d,再次亞驚厥劑量0.5μL KA複燃,選擇再次髮作級彆在Ⅳ級以上或持續狀態大鼠,且腦電圖檢測有癲癇樣波髮放者篩選為造模成功的耐藥難治性癲癇模型鼠。②分組與處理:造模成功鼠隨機分為愈癇靈榦預組、拉莫三嗪對照組、模型組,另設假手術對照組、空白對照組共計5組,每組12隻。分彆給予愈癇靈湯劑、拉莫三嗪及同等體積的蒸餾水(2 mL/d)灌胃30 d。③標本處置與檢測:採用熒光實時定量聚閤酶鏈反應(RT-PCR)的方法,分彆檢測海馬區與顳葉皮層MDR1b的錶達。結果與假手術對照組、空白對照組間比較,造模成功組海馬區MDR1b的錶達水平明顯升高(P<0.01),顳葉皮質區差異無統計學意義(P>0.05);所有造模組的海馬區MDR1b錶達高于顳葉皮質,差異有統計學意義(P<0.01)。與模型組比較,愈癇靈、拉莫三嗪榦預後能顯著降低KA緻癇鼠海馬區MDR1b的錶達水平(P<0.05);愈癇靈方組與拉莫三嗪組間比較,海馬與顳葉皮質之間差異均無統計學意義(P>0.05);各組間顳葉皮質區MDR1b錶達差異無統計學意義(P>0.05)。結論 KA造模耐藥性癲癇大鼠模型海馬區多藥耐藥基因MDR1b的錶達較皮質區明顯升高。愈癇靈方降低海馬區MDR1b的錶達,可能是其抗耐藥性癲癇作用機製之一。
목적:탐토유간령방대내약성전간모형대서해마급섭협피질다약내약기인MDR1b표체적영향。방법①조모:재뇌입체정위의인도하,해마구미량진양기주입홍조안산(KA)1.0μL(농도위1.0μg/μL)점연조모,선취발작행위급별재Racines표준Ⅳ급이상자,이병무산납급잡마서평관위간예14 d,재차아량궐제량0.5μL KA복연,선택재차발작급별재Ⅳ급이상혹지속상태대서,차뇌전도검측유전간양파발방자사선위조모성공적내약난치성전간모형서。②분조여처리:조모성공서수궤분위유간령간예조、랍막삼진대조조、모형조,령설가수술대조조、공백대조조공계5조,매조12지。분별급여유간령탕제、랍막삼진급동등체적적증류수(2 mL/d)관위30 d。③표본처치여검측:채용형광실시정량취합매련반응(RT-PCR)적방법,분별검측해마구여섭협피층MDR1b적표체。결과여가수술대조조、공백대조조간비교,조모성공조해마구MDR1b적표체수평명현승고(P<0.01),섭협피질구차이무통계학의의(P>0.05);소유조모조적해마구MDR1b표체고우섭협피질,차이유통계학의의(P<0.01)。여모형조비교,유간령、랍막삼진간예후능현저강저KA치간서해마구MDR1b적표체수평(P<0.05);유간령방조여랍막삼진조간비교,해마여섭협피질지간차이균무통계학의의(P>0.05);각조간섭협피질구MDR1b표체차이무통계학의의(P>0.05)。결론 KA조모내약성전간대서모형해마구다약내약기인MDR1b적표체교피질구명현승고。유간령방강저해마구MDR1b적표체,가능시기항내약성전간작용궤제지일。
〔Abstract〕 Objective To investigate the effects of Yuxianling Decoction (YXLD) on expression of multiple drug resistant gene MDR1b in hippocampus and temporal lobe cortex of epileptic rats induced by kaicic acid (KA). Methods (1) Making models:The Hippocampus in rats located by brain stereotactic apparatus was microinjected 1μg KA (1.0 μg/μL) to kindle seizures models. The rats at above seizure behavior surpass Ⅳ level were intragastric administration interfered by sodium valproate and carbamazepine for 14 days, and rekindled 0.5 μL. The rats second attacked seizure at above Ⅳ level and with persistent state were selected.Moreover, the rats with epileptiform discharge wave were selected to be successful resistant refractory epilepsy model rats. (2) Grouping and treatment: The Successful model rats were randomly divided into YXLD group, lamotrigine control group and model group, rats were also assigned into sham operation group and normal control group, 12 rats in each group. The rats were intragastric administration interfered by the same volume of distilled water, lamotrigine and YXLD respectively for 30 days (2 mL/d). (3) Selecting and detecting specimens: The expression of MDR 1b in hippocampus area and temporal lobe cortex was detected by real-time fluorescent quantitative polymerase chain reaction (RT-PCR) method. Results Compared with sham operation control group and normal blank control group, the expression level of MDR1b in hippocampus area was increased obviously (P<0.01), the differences in temporal lobe cortex have no statistical significance (P>0.05); The MDR1b expression in hippocampus area of all model groups was higher than that in temporal lobe cortex, the differences have statistical significance (P<0.05). Compared with the model group, lamotrigine and YXLD intervention could reduce MDR1b expression level in hippocampus area of KA epileptic rats (P<0.05); Between the YXLD group and lamotriginegroup, the differences in hippocampus and temporal lobe cortex have no statistical significance (P>0.05); MDR1b expression level intemporal lobe cortex between all groups have no statistical significance (P>0.05). Conclusion The expression of multiple drug resistant gene MDR1b in the hippocampus of KA epileptic rats was increased significantly than that in temporal lobe cortex. One of mechanisms of YXLD on anti-drug resistantepilepsy was for reversion and reducing the MDR1b expression in hippocampus area.