CAJ | 학술논문

目的探讨补阳还五汤对脑缺血后大鼠蛋白激酶B1（AKT1）和c-Jun氨基末端激酶1/2（JNK1/2）的影响。方法将48只SD大鼠按随机数字表法分为正常对照组、假手术组、模型组、补阳还五汤组，每组12只。采用大脑中动脉闭塞法（MCAO）建立右侧局灶性脑缺血大鼠模型。补阳还五汤组大鼠于术后2h起灌服补阳还五汤（主要由生黄芪120g、当归6g、赤芍4.5g、川芎3g、西红花3g、桃仁3g、广地龙3g组成）14.2 g/kg，每日1次，连续7 d。其他各组动物均给予等体积生理盐水。于第7日处死大鼠，取脑组织，采用逆转录-聚合酶链反应（RT-PCR）测定AKT1 mRNA表达，采用免疫组化法检测JNK1/2蛋白表达。结果与正常对照组和假手术组比较，模型组AKT1 mRNA表达量〔吸光度（A）值〕明显降低（0.48±0.08比0.63±0.11、0.61±0.09，均P＜0.05），JNK1/2阳性细胞数（个/mm2）明显增多（34.13±4.57比16.15±1.09、16.23±2.05，均P＜0.05）；与模型组比较，补阳还五汤组脑组织中AKT1 mRNA表达（0.93±0.11）和JNK1/2阳性细胞数（45.04±5.68）明显增加，差异均有统计学意义（P＜0.05或P＜0.01）。结论补阳还五汤可上调缺血脑组织AKT1 mRNA和JNK1/2阳性细胞数表达，是其脑保护作用机制之一。
목적탐토보양환오탕대뇌결혈후대서단백격매B1（AKT1）화c-Jun안기말단격매1/2（JNK1/2）적영향。방법장48지SD대서안수궤수자표법분위정상대조조、가수술조、모형조、보양환오탕조，매조12지。채용대뇌중동맥폐새법（MCAO）건립우측국조성뇌결혈대서모형。보양환오탕조대서우술후2h기관복보양환오탕（주요유생황기120g、당귀6g、적작4.5g、천궁3g、서홍화3g、도인3g、엄지룡3g조성）14.2 g/kg，매일1차，련속7 d。기타각조동물균급여등체적생리염수。우제7일처사대서，취뇌조직，채용역전록-취합매련반응（RT-PCR）측정AKT1 mRNA표체，채용면역조화법검측JNK1/2단백표체。결과여정상대조조화가수술조비교，모형조AKT1 mRNA표체량〔흡광도（A）치〕명현강저（0.48±0.08비0.63±0.11、0.61±0.09，균P＜0.05），JNK1/2양성세포수（개/mm2）명현증다（34.13±4.57비16.15±1.09、16.23±2.05，균P＜0.05）；여모형조비교，보양환오탕조뇌조직중AKT1 mRNA표체（0.93±0.11）화JNK1/2양성세포수（45.04±5.68）명현증가，차이균유통계학의의（P＜0.05혹P＜0.01）。결론보양환오탕가상조결혈뇌조직AKT1 mRNA화JNK1/2양성세포수표체，시기뇌보호작용궤제지일。
Objective To explore the effect of Buyang Huanwu decoction(BYHWD)on protein kinase B1 (AKT1)and c-Jun amino terminal kinase 1/2(JNK1/2)in rats after focal cerebral ischemia. Methods According to the random number table method,48 Sprague-Dawley(SD)rats were randomly allocated to four groups:normal control group,sham-operated group,model group,traditional BYHWD group(each n=12). The rat model of right focal cerebral ischemia was established by the method of middle cerebral artery occlusion(MCAO). The rats in BYHWD group were ingested with the decoction of BYHWD 14.2 g/kg after 2 hours of the operation(the main ingredients of BYHWD including astragalus mongholicus 120 g,Chinese angelica 6 g,radix paeoniae rubra 4.5 g, rhizoma ligustici wallichii 3 g,safflower 3 g,peach kernel 3 g,earthworm 3 g),once a day for 7 days. Other groups of animals were given the same amount of normal saline orally. After operation,on the 7th day,the animals were killed,and their brains were taken out. The reverse transcription-polymerase chain reaction(RT-PCR)assay was used to detect AKT1 mRNA expression,and immunohistochemical method was applied to measure JNK1/2 protein expression. Results Compared with normal control and sham-operated groups,the level of AKT1 mRNA expression〔absorbance(A)〕was decreased obviously(0.48±0.08 vs. 0.63±0.11,0.61±0.09,both P<0.05),and the number of JNK1/2 positive cells(cell/mm2)was increased significantly(34.13±4.57 vs. 16.15±1.09,16.23±2.05,both P<0.05)in model group;compared with model group,the AKT1 mRNA expression in brain tissue(0.93±0.11)and the number of JNK1/2 positive cells(45.04±5.68)was increased significantly in BYHWD group,the differences being statistically significant(P<0.05 or P<0.01). Conclusion BYHWD can up-regulate expressions of AKT1 mRNA and JNK1/2 positive cells in ischemic brain tissue that is one of the mechanisms in the protection of brain.