Neuroprotective effect of borneol combined with astragaloside Ⅳ and Panax notoginseng saponins in cerebral ischemia reperfusion injury rat model through Notch signaling pathway

OUYANG Bo1 LIU Xiao-dan1 YANG Xiao-qian1 DING Huang1 HUANG Xiao-ping1 DENG Chang-qing1

(1.Molecular Pathology Laboratory, Key Laboratory of Hunan Province for Integrated Traditional Chinese and Western Medicine on Prevention and Treatment of Cardio-Cerebral Diseases, Key Laboratory of Hunan Universities for Cell Biology and Molecular Techniques, Hunan University of Chinese Medicine, Changsha, Hunan Province, China 410208)
【Knowledge Link】optic chiasm; pyknosis; karyolysis

【Abstract】 Objective To observe the neuroprotective effect of borneol combined with astragaloside Ⅳ (AST Ⅳ) and Panax notoginseng saponins (PNS) on rat models with cerebral ischemia reperfusion injury (CIRI) from the perspective of the Notch signaling pathway. Methods SD rats were randomly divided into the sham operation group, model group, borneol (7.5 mg/kg) group, AST Ⅳ (25 mg/kg) group, PNS (10 mg/kg) group, AST Ⅳ (10 mg/kg) + PNS (25 mg/kg) group, low-dose borneol (7.5 mg/kg) + AST Ⅳ (25 mg/kg) + PNS (10 mg/kg) group, high-dose borneol (15 mg/kg) + AST Ⅳ (20 mg/kg) + PNS (50 mg/kg) group and edaravone (4 mg/kg) group. Rats in the sham operation and model groups received intragastric administration of 0.5% CMC-Na (10 mL/kg); those in the edaravone group received edaravone (2.5 mL); the ones in the remaining administration groups were treated with 2.5 mL of the corresponding drugs by gavage. The medication was provided twice per day at a 12-h interval. The right middle cerebral artery of the rat was occluded by a suture 2 h after the last administration to establish the CIRI model. After 2 h of ischemia and 22 h of reperfusion, the neurological deficit scoring was conducted and pathological changes in the ischemic cortex of rats were observed after HE staining. The protein expression levels of neuron-specific nuclear (NeuN) and endothelial barrier antigen (EBA) in the ischemic cortex were detected by immunohistochemistry. The protein expression levels of vascular endothelial growth factor (VEGF), Notch1, and the intracellular domain of Notch (NICD) in the ischemic cortex were detected by Western blotting. Results The neurological deficit score and cell damage rate in the model group were significantly increased (P < 0.01); the neurological deficit score and cell damage rate in each administration group were significantly reduced (P < 0.05, 0.01), and the effect of borneol + AST Ⅳ + PNS was better than that of any single drug or AST Ⅳ + PNS (P < 0.05, 0.01). NeuN and EBA protein expression levels were significantly decreased in the ischemic cortex of the model group (P < 0.01), while those in each administration group were significantly enhanced (P < 0.05, 0.01). The effect of borneol + AST Ⅳ + PNS was better than that of any single drug or AST Ⅳ + PNS (P < 0.05, 0.01). In the model group, VEGF protein expression was increased significantly (P < 0.05), while NICD and Notch1 protein expression levels had no significant change. The protein expression levels of VEGF, NICD, and Notch1 were significantly up-regulated in the borneol + AST Ⅳ + PNS group (P < 0.01), and the effect of the combination of three drugs was better than that of any single drug or AST Ⅳ + PNS (P < 0.05, 0.01). Conclusion Borneol, AST Ⅳ, and PNS have the effects of preventing neuronal and cerebral microvascular damage after CIRI, and the protective effect of their combination against cerebral ischemic injury was better than that of any single drug or AST Ⅳ + PNS, which might be related to the activation of the Notch signaling pathway and the up-regulation of VEGF expression.

【Keywords】 borneol; astragaloside Ⅳ; Panax notoginseng saponins; cerebral ischemia reperfusion injury; neurons; microvascular;

【DOI】

【Funds】 National Natural Science Foundation of China (81573875) Science Fund for Creative Research Groups of Natural Science in Hunan Province Science and Technology Innovation Platform and Talent Program by Science and Technology Department of Hunan Province (2017SK4005) Key Special Funding Project by Science and Technology Department of Hunan Province (2017SK2111) Natural Science Foundation of Hunan Province (2018JJ3382) Outstanding Youth Project of Education Department of Hunan Province (18B236) First-level Basic Medical Construction Project of Hunan University of Chinese Medicine during the Thirteen Five-year Plan Period (06)

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    References

    [1] Tan H, Huang X P, Deng C Q. Effects of astragalosides and Panax notoginseng saponins combination on oxidative stress of cerebral ischemic reperfusion injury in mice [J]. Journal of Integrative Medicine, 2010, 8 (5): 448–452 (in Chinese).

    [2] Huang X P, Wang P, Qiu Y Y, et al. Effects of astragaloside Ⅳ with active components in Panax notoginseng on energy metabolism in brain tissues after cerebral ischemic-reperfusion in mice [J]. Chinese Traditional and Herbal Drugs, 2014, 45 (2): 220–226 (in Chinese).

    [3] Liu X D, Yang X Q, Tang S, et al. 冰片对黄芪甲苷和三七总皂苷配伍有效成分在脑缺血/再灌注模型大鼠脑组织分布的影响 [J]. Chinese Traditional and Herbal Drugs, 2019, 50 (7): 1649–1656 (in Chinese).

    [4] Longa E Z, Weinstein P R, Carlson S, et al. Reversible middle cerebral artery occlusion without craniectomy in rats [J]. Stroke, 1989, 20 (1): 84–91.

    [5] Liu Y, Meng R, Yan F, et al. Evaluation of rat MCAO model by laser Doppler flowmetry [J]. Chinese Journal of Pathophysiology, 2011, 27 (3): 620–624 (in Chinese).

    [6] Bederson J B, Pitts L H, Tsuji M, et al. Rat middle cerebral artery occlusion: Evaluation of the model and development of a neurologic examination [J]. Stroke, 1986, 17 (3): 472–476.

    [7] Yang X Q, Chen X L, Yang R Y, et al. Study on the effective dosage of borneol combined with astragalosides Ⅳ and Panax Notoginseng saponins antagonizing cerebral ischemia-reperfusion injury [J]. Journal of Hunan University of Chinese Medicine, 2019, 39 (4): 441–447 (in Chinese).

    [8] Huang X P, Tan H, Chen B Y, et al. Combination of total astragalus extract and total Panax notoginseng saponins strengthened the protective effects on brain damage through improving energy metabolism and inhibiting apoptosis after cerebral ischemia-reperfusion in mice [J]. Chin J Integr Med, 2017, 23 (6): 445–452.

    [9] Kirino T. Delayed neuronal death [J]. Neuropathology, 2000, 20 (Sl): S95-S97.

    [10] Zhang Y H, Yao M J, Cong W H, et al. Effect of extraction of saffron crocus on mitochondrial dynamics in ischemia/reperfusion rats [J]. Chinese Pharmacological Bulletin, 2018, 34 (6): 770–775 (in Chinese).

    [11] Saubaméa B, Cochois-Guégan V, Cisternino S, et al. Heterogeneity in the rat brain vasculature revealed by quantitative confocal analysis of endothelial barrier antigen and P-glycoprotein expression [J]. J Cereb Blood Flow Metab, 2012, 32 (1): 81–92.

    [12] Omura K, Ohbayashi M, Sano M, et al. The recovery of blood-nerve barrier in crush nerve injury--a quantitative analysis utilizing immunohistochemistry [J]. Brain Res, 2004, 1001 (1/2): 13–21.

    [13] Li N, Wang P, Ma X L, et al. Effect of bone marrow stromal cell transplantation on neurologic function and expression of VEGF in rats with focal cerebral ischemia [J]. Mol Med Rep, 2014, 10 (5): 2299–305.

    [14] Pan Z G, Mao Y, Sun F Y. VEGF enhances reconstruction of neurovascular units in the brain after injury [J]. Acta Physiologica Sinica, 2017, 69 (1): 96–108 (in Chinese).

    [15] Jiang H H, Wang Y Y, Hu D H, et al. Effects of Panax Notoginseng Saponin on the Expression of Vascular Endothelial Growth Factor after the Brain Ischemia-reperfusion Injury in Rats [J]. Journal of Yangtze University (Natural Science Edition), 2015, 12 (12): 1–5 (in Chinese).

    [16] Zhang Q, Li M L, Zhao Y, et al. 黄芪加冰片对大鼠脑缺血再灌注损伤后VEGF表达的影响 [J]. Shandong Medical Journal, 2010, 50 (7): 88–89 (in Chinese).

    [17] Hayase M, Kitada M, Wakao S, et al. Committed neural progenitor cells derived from genetically modified bone marrow stromal cells ameliorate deficits in a rat model of stroke [J]. J Cereb Blood Flow Metab, 2009, 29 (8): 1409–1420.

    [18] Zeng H Y, Peng Y J, Wang Y. Role of Notch-1/NICD signaling in probucol's protection on rats with acute cerebral ischemia reperfusion injury and its mechanism [J]. Chinese Journal of Anatomy, 2016, 39 (6): 656–659 (in Chinese).

    [19] Liu Q, Wei L S, Wang Y, et al. Notch信号通路在大鼠骨髓间充质干细胞移植后促进脑缺血区血管新生过程中的作用 [J]. Chinese Journal of Gerontology, 2017, 37 (17): 4185–4187 (in Chinese).

    [20] Song Z Y, Zhou Y, Wang S S, et al. Experimental Research of Naotaifang on Expression of VEGF and Angiopoietins in Rats after Cerebral Ischemia [J]. Journal of Hunan University of Chinese Medicine, 2016, 36 (1): 19–22 (in Chinese).

This Article

ISSN:0253-2670

CN: 12-1108/R

Vol 51, No. 23, Pages 5990-5997

December 2020

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Abstract

  • 1 Materials
  • 2 Methods
  • 3 Results
  • 4 Discussion
  • References