3-乙酰基-11-酮-β乳香酸对大鼠脑缺血再灌注损伤的保护作用及其机制研究

doi:10.3877/cma.j.issn.1674-6880.2021.06.005

目的

探讨乳香提取物3-乙酰基-11-酮-β-乳香酸（AKBA）对大鼠脑缺血再灌注损伤的保护作用及其机制。

方法

将80只大鼠分为假手术组、模型组、低剂量组及高剂量组，每组各20只大鼠。除假手术组外，其他三组采用改良线栓方法制备大鼠大脑中动脉缺血再灌注损伤模型，低剂量组在造模成功再灌注后60 min给予腹腔注射10 mg / kg的AKBA，高剂量组则给予20 mg / kg的AKBA。测定并计算各组大鼠脑梗死率及梗死区细胞凋亡率；采用转录实时荧光定量PCR检测微RNA-320（miRNA-320）和胰岛素样生长因子1（IGF-1）的信使RNA（mRNA）表达水平，采用Western-blotting测定脑组织IGF-1的蛋白表达情况。

结果

各组大鼠脑梗死率、细胞凋亡率、miRNA-320、IGF-1的mRNA及蛋白表达水平比较，差异均有统计学意义（F = 193.300、230.000、100.900、180.400、26.180，P均< 0.001）。进一步两两比较发现，与假手术组比较，模型组大鼠的细胞凋亡率、miRNA-320、IGF-1的mRNA及蛋白表达水平均显著升高（P均< 0.05）；与模型组比较，低剂量组及高剂量组大鼠的脑梗死率、细胞凋亡率及miRNA-320表达水平均显著降低，IGF-1的mRNA及蛋白表达水平均显著升高（P均< 0.05）。

结论

AKBA对大鼠脑缺血再灌注损伤有明显的保护作用,其机制可能是通过抑制miRNA-320的表达水平及上调IGF-1的水平来减少神经细胞的凋亡。

关键词: 再灌注损伤, 脑缺血, 3-乙酰基-11-酮-β乳香酸, 大鼠

Objective

To investigate the protective effect and mechanism of 3-acetyl-11-keto-β-boswellic acid (AKBA) on cerebral ischemia-reperfusion injury in rats.

Methods

A total of 80 rats were randomly divided into a sham operation group, a model group, a low-dose group and a high-dose group, with 20 rats in each group. Except for the sham operation group, a middle cerebral artery ischemia-reperfusion injury model was induced by a modified suture method in the other three groups. The rats in the low-dose group were given an intraperitoneal injection of 10 mg / kg AKBA at 60 min after successful reperfusion, while the rats in the high-dose group were given 20 mg / kg AKBA. The rates of cerebral infarction and apoptosis were measured and calculated in the four groups. The messenger RNA (mRNA) expression of microRNA-320(miRNA-320) and insulin-like growth factor-1 (IGF-1) was detected by reverse transcription real-time fluorescence quantitative PCR, and the protein expression of IGF-1 was measured by Western-blotting.

Results

The cerebral infarction rate, apoptosis rate, miRNA-320 mRNA, IGF-1 mRNA and IGF-1 protein among the four groups all showed significant differences (F = 193.300, 230.000, 100.900, 180.400, 26.180; all P < 0.001). Further pairwise comparison revealed that the apoptosis rate, miRNA-320 mRNA, IGF-1 mRNA and IGF-1 protein in the model group were much higher than those in the sham operation group (all P < 0.05). The rates of cerebral infarction and apoptosis and the expression of miRNA-320 mRNA were much lower, while the expression of IGF-1 mRNA and protein was much higher in the low-dose group and high-dose group than in the model group (all P < 0.05).

Conclusion

AKBA has an obvious protective effect on cerebral ischemia-reperfusion injury in rats, and it may be to reduce neuronal apoptosis by inhibiting miRNA-320 and up-regulating IGF-1.

Key words: Reperfusion injury, Brain ischemia, 3-acetyl-11-keto-β-boswellic acid, Rats

图1 TTC染色下各组大鼠脑组织梗死情况（n = 5）注：TTC.氯化三苯基四氮唑；a图为各组大鼠脑组织梗死情况，正常组织呈红色，梗死组织呈白色；b图为各组大鼠脑梗死率的比较，与模型组比较，^aP < 0.05

表1 各组大鼠脑组织细胞凋亡率的比较（%， ± s）

组别	只数	细胞凋亡率
假手术组	5	2.9 ± 0.5
模型组	5	22.4 ± 2.7^a
低剂量组	5	19.6 ± 1.0^ab
高剂量组	5	7.0 ± 0.9^ab
F值		230.000
P值		< 0.001

表1 各组大鼠脑组织细胞凋亡率的比较（%， ± s）

组别	只数	细胞凋亡率
假手术组	5	2.9 ± 0.5
模型组	5	22.4 ± 2.7^a
低剂量组	5	19.6 ± 1.0^ab
高剂量组	5	7.0 ± 0.9^ab
F值		230.000
P值		< 0.001

图2 各组大鼠脑组织检测细胞凋亡表达情况（TUNEL染色　× 400）注：TUNEL.脱氧核苷酸末端转移酶介导的核苷酸缺口末端标记法；TUNEL染色后正常细胞核呈蓝色，阳性凋亡细胞核呈绿色；a图为假手术组，可见少许凋亡细胞；b图为模型组，可见大量阳性凋亡细胞；c ~ d图分别为低剂量组和高剂量组，可见阳性凋亡细胞数量明显下降

图3 各组大鼠脑组织中miRNA-320及IGF-1 mRNA表达水平的比较（n = 5）注：miRNA-320.微RNA-320；IGF-1.胰岛素样生长因子1；mRNA.信使RNA；GAPDH. 3-磷酸甘油醛脱氢酶；与假手术组比较，^aP < 0.05；与模型组比较，^bP < 0.05

图4 各组大鼠脑组织中IGF-1的蛋白表达情况（n = 5）注：IGF-1.胰岛素样生长因子1；GAPDH. 3-磷酸甘油醛脱氢酶；与假手术组比较，^aP < 0.05；与模型组比较，^bP < 0.05

1	施成彰，沈建，詹仁雅. 脑缺血再灌注损伤与还原型烟酰胺腺嘌呤二核苷酸磷酸氧化酶的关系及其研究进展[J/CD]. 中华危重症医学杂志（电子版），2017，10(6):411-415.
2	Hamidpour R, Hamidpour S, Hamidpour M, et al. Fr-ankincense (ru xiang; boswellia species): from the selection of traditional applications to the novel phytotherapy for the prevention and treatment of serious diseases[J]. J Tradit Complement Med, 2013, 3 (4): 221-226.
3	Yu G, Xiang W, Zhang T, et al. Effectiveness of Boswellia and Boswellia extract for osteoarthritis patients: a systematic review and meta-analysis[J]. BMC Complement Med Ther, 2020, 20 (1): 225.
4	Ranzato E, Martinotti S, Volante A, et al. The major Boswellia serrata active 3-acetyl-11-keto-β-boswellic acid strengthens interleukin-1α upregulation of matrix metalloproteinase-9 via JNK MAP kinase activation[J]. Phytomedicine, 2017 (36): 176-182.
5	Shang P, Liu W, Liu T, et al. Acetyl-11-keto-β-bo-swellic acid attenuates prooxidant and profibrotic mechanisms involving transforming growth factor-β1, and improves vascular remodeling in spontaneously hypertensive rats[J]. Sci Rep, 2016 (6): 39809.
6	刘迪，张冰洋，姚铁，等. 乳香化学成分及其药理作用研究进展[J]. 中草药，2020，51(22):5900-5914.
7	Kruger P, Daneshfar R, Eckert GP, et al. Metabolism of boswellic acids in vitro and in vivo[J]. Drug Metab Dispos, 2008, 36 (6): 1135-1142.
8	Gong L, Tang Y, An R, et al. RTN1-C mediates ce-rebral ischemia / reperfusion injury via ER stress and mitochondria-associated apoptosis pathways[J]. Cell Death Dis, 2017, 8 (10): e3080.
9	周密，张明瑜，许晓东，等. 乌司他丁预先给药对大鼠海马CA1区缺血-再灌注损伤的干预研究[J/CD]. 中华危重症医学杂志（电子版），2015，8(1):24-28.
10	Hayes CA, Valcarcel-Ares MN, Ashpol NM. Preclinical and clinical evidence of IGF-1 as a prognostic marker and acute intervention with ischemic stroke[J]. J Cereb Blood Flow Metab, 2021, 41 (10): 2475-2491.
11	Wrigley S, Arafa D, Tropea D. Insulin-like growth factor 1: at the crossroads of brain development and aging[J]. Front Cell Neurosci, 2017 (11): 14.
12	Dyer AH, Vahdatpour C, Sanfeliu A, et al. The role of insulin-like growth factor 1 (IGF-1) in brain development, maturation and neuroplasticity[J]. Neuroscience, 2016 (325): 89-99.
13	Sohrabji F. Estrogen-IGF-1 interactions in neuroprotec-tion: ischemic stroke as a case study[J]. Front Neuroendocrinol, 2015 (36): 1-14.
14	Lu Y, Sareddy GR, Wang J, et al. Neuron-derived estrogen is critical for astrocyte activation and neuroprotection of the ischemic brain[J]. J Neurosci, 2020, 40 (38): 7355-7374.
15	Gong P, Zou Y, Zhang W, et al. The neuroprotective effects of insulin-like growth factor 1 via the Hippo / YAP signaling pathway are mediated by the PI3K / AKT cascade following cerebral ischemia / reperfusion injury[J]. Brain Res Bull, 2021 (177): 373-387.
16	Bake S, Okoreeh AK, Alaniz RC, et al. Insulin-like growth factor (IGF)-Ⅰ modulates endothelial blood-brain barrier function in ischemic middle-aged female rats[J]. Endocrinology, 2016, 157 (1): 61-69.
17	Zhao B, Zheng Z. Insulin growth factor 1 protects neural stem cells against apoptosis induced by hypoxia through Akt / mitogen-activated protein kinase / extracellular signal-regulated kinase (Akt / MAPK / ERK) pathway in hypoxia-ishchemic encephalopathy[J]. Med Sci Monit, 2017 (23): 1872-1879.
18	Parker K, Berretta A, Saenger S, et al. PEGylated insulin-like growth factor-Ⅰ affords protection and facilitates recovery of lost functions post-focal ischemia[J]. Sci Rep, 2017, 7 (1): 241.
19	Rajman M, Schratt G. MicroRNAs in neural develop-ment: from master regulators to fine-tuners[J]. Development, 2017, 144 (13): 2310-2322.
20	Zhai F, Zhang X, Guan Y, et al. Expression profiles of microRNAs after focal cerebral ischemia / reperfusion injury in rats[J]. Neural Regen Res, 2012, 7 (12): 917-923.
21	Song CL, Liu B, Diao HY, et al. Down-regulation of microRNA-320 suppresses cardiomyocyte apoptosis and protects against myocardial ischemia and reperfusion injury by targeting IGF-1[J]. Oncotarget, 2016, 7 (26): 39740-39757.
22	梁丽贞，王俊，严永兴，等. microRNA-320对小鼠脑缺血-再灌注损伤的调控作用及其机制研究[J]. 中华急诊医学杂志，2018，27(3):313-317.
23	Toden S, Okugawa Y, Buhrmann C, et al. Novel evidence for curcumin and boswellic acid-induced chemoprevention through regulation of miR-34a and miR-27a in colorectal cancer[J]. Cancer Prev Res (Phila), 2015, 8 (5): 431-443.

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Protective effect and mechanism of 3-acetyl-11-keto-β-boswellic acid on cerebral ischemia-reperfusion injury in rats

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Protective effect and mechanism of 3-acetyl-11-keto-β-boswellic acid on cerebral ischemia-reperfusion injury in rats