CAJ | 학술논문

目的：探讨黄芩苷对高原性脑缺血缺氧致小鼠脑损伤的影响及相关作用靶点蛋白表达。方法：用water maze水迷宫作业测试筛选出50只昆明种小鼠，随机分为模型组、对照组、黄芩苷低、中、高剂量组（0.05、0.20、0.60 mg·kg-1），每组10只，检测受试小鼠的空间记忆和学习能力。用低压氧动物舱（模拟海拔4000 m的高度）建立稳定的高原性脑缺血缺氧模型，检测小鼠海马组织中超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-PX）活性及丙二醛（MDA）含量的变化。应用蛋白印迹分析方法（western blot）分别检测小鼠脑干组织中各凋亡通路蛋白活化型半胱天冬酶-3（cleaved-caspase 3）、磷酸化蛋白激酶B（P-AKT）、神经胶质酸性蛋白（GFAP）、Bax、Bcl-2的表达。结果：模型组小鼠潜伏期明显高于对照组（P<0.05），黄芩苷高剂量组与模型组相比潜伏期缩短，差异显著（P<0.05），由此得出，黄芩苷最佳有效剂量为0.60 mg·kg-1。与对照组比较，模型组小鼠海马组织中MDA含量明显升高、SOD和GSH-Px活性明显降低（P<0.05）；与模型组比较，黄芩苷高剂量组小鼠脑组织中SOD和GSH-Px活性明显提高、MDA含量明显降低（P<0.05）。从蛋白变化水平来看，模型组的cleaved-caspase 3、P-AKT、GFAP蛋白表达水平与对照组相比，条带明显增强，Bax/Bcl-2的比值也明显增加（P<0.05）；黄芩苷组表达水平弱于模型组（P<0.05），其中黄芩苷高剂量组表达最弱，且呈一定的量效关系。结论：黄芩苷对高原性脑缺血损伤有保护作用，其作用机制可能与强的抗氧化能力及其抑制各个靶点蛋白cleaved-caspase 3、P-AKT、GFAP、Bax、Bcl-2在凋亡通路的表达有关。
목적：탐토황금감대고원성뇌결혈결양치소서뇌손상적영향급상관작용파점단백표체。방법：용water maze수미궁작업측시사선출50지곤명충소서，수궤분위모형조、대조조、황금감저、중、고제량조（0.05、0.20、0.60 mg·kg-1），매조10지，검측수시소서적공간기억화학습능력。용저압양동물창（모의해발4000 m적고도）건립은정적고원성뇌결혈결양모형，검측소서해마조직중초양화물기화매（SOD）、곡광감태과양화물매（GSH-PX）활성급병이철（MDA）함량적변화。응용단백인적분석방법（western blot）분별검측소서뇌간조직중각조망통로단백활화형반광천동매-3（cleaved-caspase 3）、린산화단백격매B（P-AKT）、신경효질산성단백（GFAP）、Bax、Bcl-2적표체。결과：모형조소서잠복기명현고우대조조（P<0.05），황금감고제량조여모형조상비잠복기축단，차이현저（P<0.05），유차득출，황금감최가유효제량위0.60 mg·kg-1。여대조조비교，모형조소서해마조직중MDA함량명현승고、SOD화GSH-Px활성명현강저（P<0.05）；여모형조비교，황금감고제량조소서뇌조직중SOD화GSH-Px활성명현제고、MDA함량명현강저（P<0.05）。종단백변화수평래간，모형조적cleaved-caspase 3、P-AKT、GFAP단백표체수평여대조조상비，조대명현증강，Bax/Bcl-2적비치야명현증가（P<0.05）；황금감조표체수평약우모형조（P<0.05），기중황금감고제량조표체최약，차정일정적량효관계。결론：황금감대고원성뇌결혈손상유보호작용，기작용궤제가능여강적항양화능력급기억제각개파점단백cleaved-caspase 3、P-AKT、GFAP、Bax、Bcl-2재조망통로적표체유관。
This study was aimed to explore the mechanism of baicalin on high altitude cerebral hypoxia-ischemia on mice and its influence on related target protein expressions. Morris water maze was used to screen 50 Kunming mice, which were randomly divided into the model group, control group, the low dose (0.05 mg·kg-1), middle dose (0.20 mg·kg-1) and high dose (0.60 mg·kg-1) baicalin group, with 10 rats in each group. The space memory and learning ability of mice were tested. The animal cabin with low oxygen (simulating at 4 000 m altitude) was used to establish the stable high altitude cerebral hypoxia-ischemia mouse model. Changes on SOD content, GSH-PX activities and MDA content in hippocampal tissues of mice were detected. The expressions of different target proteins, including cleaved-caspase 3, P-AKT, GFAP, Bax and Bcl-2 in brain stem of mice were detected by western blot. The results showed that the latent period of the model group was obviously longer than that of the control group (P < 0.05). The latent period of high dose baicalin group was shorter than the model group with significant difference (P< 0.05). Therefore, the best effective dose of baicalin was 0.60 mg·kg-1. Compared with the control group, the content of MDA in the hippocampal tissues of mice in the model group was significantly increased; the SOD and GSH-PX activity were obviously reduced (P < 0.05). Compared with the model group, the SOD and GSH-PX activity were obviously increased in the brain tissues of mice in the high dose baicalin group; and the content of MDA was obviously reduced (P < 0.05). From the level of protein changes, the stripes of cleaved-caspase 3, P-AKT, GFAP protein expressions in the model group were strengthened compared to the control group; the ratio of Bax/Bcl-2 was also obviously increased (P < 0.05). The expression of the baicalin group was lower than that of the model group (P < 0.05). Among them, the expression of the high dose baicalin group was the lowest. It had certain dose-response relationship. It was concluded that baicalin had protective effect on high altitude cerebral hypoxia-ischemia. Its mechanism may be related to its powerful oxidation resistance and its inhibition on expression of different target proteins, including cleaved-caspase 3, P-AKT, GFAP, Bax, Bcl-2 for the change of apoptotic pathway.