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广州中医药大学 基础医学院,广州 510006
[第一作者] 杨开令,在读硕士,从事中医药防治心脑血管疾病的研究,E-mail:2685453254@qq.com
*刘微,博士,教授,从事中医药防治心脑血管疾病的研究,E-mail:weiliu1980@yahoo.com
收稿日期:2019-04-15,
网络出版日期:2019-08-07,
纸质出版日期:2020-01-05
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杨开令, 周颖, 闫福曼, 等. 补阳还五汤对脑缺血再灌注大鼠恢复期突触可塑性的影响[J]. 中国实验方剂学杂志, 2020,26(1):43-49.
Kai-ling YANG, Ying ZHOU, Fu-man YAN, et al. Effect of Buyang Huanwu Tang on Synaptic Structural Plasticity After Cerebral Ischemia-reperfusion in Rats[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(1): 43-49.
杨开令, 周颖, 闫福曼, 等. 补阳还五汤对脑缺血再灌注大鼠恢复期突触可塑性的影响[J]. 中国实验方剂学杂志, 2020,26(1):43-49. DOI: 10.13422/j.cnki.syfjx.20192201.
Kai-ling YANG, Ying ZHOU, Fu-man YAN, et al. Effect of Buyang Huanwu Tang on Synaptic Structural Plasticity After Cerebral Ischemia-reperfusion in Rats[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(1): 43-49. DOI: 10.13422/j.cnki.syfjx.20192201.
目的:
2
探讨补阳还五汤在脑缺血再灌注大鼠恢复期提高突触可塑性的机制研究。
方法:
2
建立大脑中动脉栓塞(MCAO)大鼠模型,随机分为假手术组、模型组、补阳还五汤组、补阳还五汤联合缝隙连接蛋白43(Cx43)抑制剂(Gap26)组,补阳还五汤每天灌胃2次(16 g·kg
-1
),Gap26于术后第3天腹腔注射,每天1次(25 μg·kg
-1
);7 d后取材,采用透射电镜观察缺血侧海马突触和缝隙连接超微结构的改变,运用蛋白免疫印迹法(Western blot)和免疫荧光检测缺血侧海马突触素(SYN),生长相关蛋白-43(GAP-43)的表达。
结果:
2
电镜下观察到假手术组突触结构完整、清晰,突触数量多,缝隙连接结构清晰;模型组缺血侧海马突触结构溶解,突触数量减少,缝隙连接消失,存在较大间隙,与假手术组比较,SYN,GAP-43的表达明显增高(
P
<
0.05,
P
<
0.01);补阳还五汤组缺血侧海马突触结构较清晰,突触数量增多,缝隙连接结构较完整,与模型组比较,SYN,GAP-43的表达明显增强(
P
<
0.05,
P
<
0.01);而联合使用Gap26后缺血侧海马突触数量较补阳还五汤组减少,仅可见少量结构完整的缝隙连接,补阳还五汤增强SYN,GAP-43的作用被明显抑制(
P
<
0.05,
P
<
0.01)。
结论:
2
补阳还五汤可提高脑缺血再灌注恢复期缺血侧海马突触可塑性,其机制可能与增加Cx43的表达促进对SYN,GAP-43的干预有关。
Objective:
2
To investigate the mechanism of Buyang Huanwu Tang (BYHWT) in improving synaptic structural plasticity after cerebral ischemia-reperfusion in rats.
Method:
2
Middle cerebral artery occlusion and reperfusion model was established. SD rats were randomly divided into sham-operated group
model group
BYHWT group
BYHWT+ Gap26(connexin43 inhibitor)groups. BYHWT was given twice a day(16 g·kg
-1
)
Gap26 was intraperitoneally injected once a day since the third day after surgery (25 g·kg
-1
). Brain was taken out at the 7
th
day. The changes of neuronal synaptic and gap junction ultrastructure were observed by transmission electron microscopy. Synaptophysin (SYN) and growth-associated protein-43 (GAP-43) protein expression were detected by Western blot and immunofluorescence.
Result:
2
The structure of synapses was integrated
and the gap junctions were clear in sham-operated group. In the hippocampus of model group
the structure was destroyed
and the gap junctions disappeared. Compared with the sham-operated group
model group up-regulated the expressions of SYN and GAP-43 (
P
<
0.05
P
<
0.01). In the hippocampus of BYHWT group
the structure was close to the normal. Furthermore
BYHWT up-regulated the expressions of SYN and GAP-43 (
P
<
0.05
P
<
0.01). However
after the combined administration with Cx43 inhibitor (Gap26)
the damage of synaptic structural decreased
only a small number of gap junctions with the structural integrity can be seen
and the effect of BYHWT on SYN and GAP-43 was inhibited (
P
<
0.05
P
<
0.01).
Conclusion:
2
BYHWT could improve the hippocampal synaptic structural plasticity obviously after the CIRI. The mechanism may be related to the increase of the expression of Cx43 and the promotion of the intervention of SYN and GAP-43.
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