巴佛洛霉素A1介导AMPK/mTOR/ULK1信号通路对脑缺血再灌注损伤小鼠的神经保护机制研究

周超瑞; 顾健坤; 陈娜; 李明娟

doi:10.16766/j.cnki.issn.1674-4152.003371

巴佛洛霉素A1介导AMPK/mTOR/ULK1信号通路对脑缺血再灌注损伤小鼠的神经保护机制研究

doi: 10.16766/j.cnki.issn.1674-4152.003371

1.
海警医院麻醉科，浙江嘉兴 314000
2.
嘉兴学院医学院，浙江嘉兴 314001

基金项目:

浙江省基础公益研究项目 LGD21H300001

详细信息

通讯作者:
周超瑞，E-mail: 914897113@qq.com

中图分类号: R743.31 R-332
计量
- 文章访问数: 265
- HTML全文浏览量: 100
- PDF下载量: 16
- 被引次数: 0
出版历程
- 收稿日期: 2023-11-14
- 网络出版日期: 2024-03-27

Neuroprotective effect of Bafilomycin A1 on cerebral ischaemia reperfusion injury by regulation AMPK/mTOR/ULK1 signaling pathway in mice

1.
Department of Anesthesiology, China Coast Guard Hospital of the People's Armed Police Force, Jiaxing, Zhejiang 314000, China

摘要

摘要: 目的探讨巴佛洛霉素A1(BAF-A1)抗小鼠脑缺血再灌注(IR)损伤的腺苷一磷酸活化蛋白激酶(AMPK)/哺乳动物雷帕霉素靶蛋白(mTOR)/Unc-51样激酶1(ULK1)自噬信号通路的机制。方法选择SPF级雄性小鼠40只，采用随机数表法分成假手术组(S组)、IR模型组(M组)、BAF-A1组(B组)、BAF-A1+AMPK激动剂组(B+M组)，每组10只。以线栓法制备IR模型，以缺血1.5 h再灌注24 h后为评价点进行评价。结果神经功能缺失评分显示，S组无神经功能缺失，B组为(1.47±0.28)分、B+M组为(2.05±0.16)分、M组为(2.45±0.31)分，B组及B+M组均低于M组(P＜0.05)。与M组比较，B组脑梗死体积比率下降(P＜0.05)。与S组比较，M组CA1、CA3区有少量的核固缩核裂变、坏死的现象；与M组比较，B组CA1、CA3区的核固缩，核裂变空泡及坏死的症状改善，但B+M组拮抗了这一作用。与S组比较，M组LC3Ⅱ/Ⅰ、p-AMPK/AMPK蛋白均升高(P＜0.05)；与M组比较，B组LC3Ⅱ/Ⅰ、p-AMPK/AMPK蛋白均下降(P＜0.05)。与S组比较，M组p-mTOR/mTOR、Ps757-ULK1/ULK1蛋白降低；与M组比较，B组p-mTOR/mTOR、Ps757-ULK1/ULK1蛋白均升高(P＜0.05)。结论 BAF-A1可能通过抑制AMPK/mTOR/ULK1信号通路，抑制小鼠脑海马区的过度自噬，从而发挥神经功能的保护性作用。
- 巴佛洛霉素A1 /
- 脑缺血再灌注损伤 /
- 腺苷一磷酸活化蛋白激酶 /
- 雷帕霉素靶蛋白 /
- Unc-51样激酶1 /
- 神经保护
Abstract: Objective To investigate the mechanism by which bafilomycin A1 (BAF-A1) attenuates cerebral ischemia-reperfusion (IR) injury in mice by regulating autophagy through the adenosine activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR)/UNC-51 like kinase 1 (ULK1) signaling pathway. Methods A total of 40 SPF grade male mice were select for the experiment and randomly divided into sham group (S group), model group (M group), BAF-A1 group (B group) and BAF-A1+AMPK group (B+M group) group. An IR model is prepared using the suture method and evaluated using 1.5 hours of ischaemia followed by 24 hours of reperfusion as the observation point. Results There was no neurological deficit in S group mice, the scores of B Group (1.47±0.28) and Group B+M (2.05±0.16) decreased compared with M Group (2.45±0.31), P < 0.05. Compared with S Group, there was significant left cerebral ischemia in M Group, B Group and B+M Group, with white infarcted areas visible. Compared with M group, the volume ratio of cerebral infarction in B group mice was significantly reduced (P < 0.05). Compared with S group mice, M group mice showed a small amount of nuclear pyknosis and fission in the neurons of the CA1 and CA3 regions of the hippocampus. Compared with M group mice, group B mice showed enhanced nuclear pyknosis, nuclear fission vacuoles, and necrosis in the CA1 and CA3 regions, but the B+M group antagonized this effect. Compared with S group, LC3 Ⅱ/Ⅰ, p-AMPK/AMPK of ischaemic brain tissue in M group rats increased (P < 0.05); compared with M group, B Group showed a decrease in LC3 Ⅱ/Ⅰ, p-AMPK/AMPK (P < 0.05). Compared with S group, the p-mTOR/mTOR and Ps757-ULK1/ULK1 ratio decreased in M group; compared with M group, the p-mTOR/mTOR and Ps757-ULK1/ULK1 ratio were significantly increased in B group (P < 0.05). Conclusion BAF-A1 pretreatment may exert a protective effect on neural function by inhibiting the AMPK/mTOR/ULK1 signaling pathway, thereby inhibiting excessive autophagy in the hippocampus of mice.
- Bafilomycin A1 /
- Cerebral ischemia-reperfusion injury /
- Adenosine activated protein kinase /
- Mammalian target of rapamycin /
- UNC-51 like kinase 1 /
- Neuroprotection

HTML全文

图 1 BAF-A1对脑梗死体积的影响

注：与S组比较，^aP＜0.05；与M组比较，^bP＜0.05；与B组比较，^cP＜0.05。

Figure 1. Effect of BAF-A1 on the volume of cerebral infarction

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图 2 BAF-A1对小鼠海马区病理形态学变化的影响(HE染色，×400)

Figure 2. Effect of BAF-A1 on pathological and morphological changes in the hippocampus of mice (HE staining, ×400)

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图 3 BAF-A1对相关蛋白的表达影响

Figure 3. Effects of BAF-A1 on the expression of related proteins

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表 1 BAF-A1对小鼠神经功能缺失评分的影响(x±s，分)

Table 1. Effect of BAF-A1 on neurological deficit score in mice (x±s, points)

组别	只数	神经功能缺失评分
S组	10	0
M组	10	2.45±0.31^a
B组	10	1.47±0.28^b
B+M组	10	2.05±0.16^bc
F值		36.403
P值		＜0.001
注：与S组比较, ^aP＜0.05;与M组比较，^bP＜0.05;与B组比较, ^cP＜0.05。

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表 2 BAF-A1对相关蛋白表达的影响(x±s)

Table 2. Effects of BAF-A1 on the expression of related proteins(x±s)

组别	只数	LC3Ⅱ/Ⅰ	p-AMPK/AMPK	p-mTOR/mTOR	Ps757-ULK1/ULK1
S组	3	0.42±0.05	0.47±0.07	0.88±0.10	1.12±0.13
M组	3	0.61±0.09^a	0.84±0.07^a	0.60±0.07^a	0.64±0.09^a
B组	3	0.45±0.06^b	0.53±0.08^b	0.78±0.09^b	0.92±0.11^b
B+M组	3	0.74±0.05^c	0.96±0.06^c	0.51±0.08^bc	0.61±0.13^c
F值		15.931	34.142	11.520	13.071
P值		0.001	＜0.001	0.003	0.002
注：与S组比较, ^aP＜0.05；与M组比较，^bP＜0.05；与B组比较, ^cP＜0.05。

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