羽扇豆醇对脑缺血再灌注大鼠氧化应激损伤和炎性反应的调节作用及机制研究
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摘要
目的探讨羽扇豆醇对大鼠脑缺血再灌注氧化应激和炎性反应的影响。方法 120只SD大鼠随机分为假手术组、模型组、不同浓度(10、20、50 mg/kg)羽扇豆醇组。模型组和不同浓度羽扇豆醇组利用改良线栓法制备大鼠脑缺血再灌注损伤模型。苏木素伊红染色检测5组大鼠脑组织病理损伤情况,TUNEL染色检测脑组织细胞凋亡情况,蛋白免疫印迹检测Bax和Bcl-2表达情况,酶联免疫吸附试验检测血清氧化应激标记物水平和炎性因子水平。结果与模型组比较,不同浓度羽扇豆醇组脑组织损伤程度减轻,细胞凋亡率、Bax表达水平降低,Bcl-2蛋白表达水平升高(P <0. 01)。与模型组比较,不同浓度羽扇豆醇组超氧化物歧化酶和还原型谷胱甘肽水平升高,丙二醛水平和炎性因子水平降低(P <0. 01)。结论羽扇豆醇可改善大鼠脑缺血再灌注氧化应激损伤,并减轻炎性反应。
Objective To investigate effects of Lupeol on oxidative stress injuries and inflammatory reaction in rats with cerebral ischemia-reperfusion. Methods A total of 120 SD rats were randomly divided into control group,model group and different Lupeol concentrations groups(10,20 and 50 mg/Kg). Models of cerebral ischemia-reperfusion were prepared by using modified middle cerebral artery occlusion(MCAO) method. In five groups,histopathological changes of brain tissues were detected by hematoxylin-feosin staining; apoptosis rates of brain tissue cells were detected by TUNEL staining; protein expressions of Bax and Bcl-2 were detected by western-blot,and serum levels of oxidative stress markers and inflammatory cytokines were detected by enzyme linked immunosorbent assay(ELISA). Results Compared with those in model group,in different Lupeol concentrations groups,damage degrees of brain tissues were decreased,and apoptosis rates and expressions of Bax were reduced,while expressions of Bcl-2 protein were increased(P < 0. 01); levels of superoxide dismutase(SOD) and glutathione(GSH) were increased,while levels of malondialdehyde(MDA) and inflammatory factors were decreased(P < 0. 01). Conclusion Lupeol may ameliorate oxidative stress injuries and decrease inflammatory reactions in rats with cerebral ischemia-reperfusion.
Objective To investigate effects of Lupeol on oxidative stress injuries and inflammatory reaction in rats with cerebral ischemia-reperfusion. Methods A total of 120 SD rats were randomly divided into control group,model group and different Lupeol concentrations groups(10,20 and 50 mg/Kg). Models of cerebral ischemia-reperfusion were prepared by using modified middle cerebral artery occlusion(MCAO) method. In five groups,histopathological changes of brain tissues were detected by hematoxylin-feosin staining; apoptosis rates of brain tissue cells were detected by TUNEL staining; protein expressions of Bax and Bcl-2 were detected by western-blot,and serum levels of oxidative stress markers and inflammatory cytokines were detected by enzyme linked immunosorbent assay(ELISA). Results Compared with those in model group,in different Lupeol concentrations groups,damage degrees of brain tissues were decreased,and apoptosis rates and expressions of Bax were reduced,while expressions of Bcl-2 protein were increased(P < 0. 01); levels of superoxide dismutase(SOD) and glutathione(GSH) were increased,while levels of malondialdehyde(MDA) and inflammatory factors were decreased(P < 0. 01). Conclusion Lupeol may ameliorate oxidative stress injuries and decrease inflammatory reactions in rats with cerebral ischemia-reperfusion.
引文
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[14]谭永星,袁楠楠,夏裕宁,等. H2对大鼠全脑缺血再灌注损伤后海马CA1区神经元及树突棘的保护作用[J].中华神经医学杂志,2016,15(11):1091-1097.
[15] Sunitha S,Nagaraj M,Varalakshmi P. Hepatoprotective effect of lupeol and lupeol linoleate on tissue antioxidant defence system in cadmium-induced hepatotoxicity in rats[J]. Fitoterapia,2001,72(5):516-523.
[16] Prasad S,Kalra N,Shukla Y. Hepatoprotective effects of lupeol and mango pulp extract of carcinogen induced alteration in swiss albino mice[J]. Mol Nutr Food Res,2007,51(3):352-359.
[17] Yamashita K,Lu H,Lu J,et al. Effect of three triterpenoids,lupeol,betulin,and betulinic acid on the stimulus-induced superoxide generation and tyrosyl phosphorylation of proteins in human neutrophils[J]. Clin Chim Acta,2002,325(1-2):91-96.
[18] Prasad S,Kalra N,Singh M,et al. Protective effects of lupeol and mango extract against androgen induced oxidative stress in swiss albino mice[J]. Asian J Androl,2008,10(2):313-318.
[19] Sieber M W,Claus R A,Witte O W,et al. Attenuated inflammatory response in aged mice brains following stroke[J]. PLo S One,2011,6(10):e26288.
[20]孙恺,张秀萍,宋林杰,等.拮抗高迁移率族蛋白B1对糖尿病合并脑缺血再灌注损伤的保护作用研究[J].中华神经医学杂志,2016,15(9):901-907.
[21] Dziedzic T. Systemic inflammation as a therapeutic target in acute ischemic stroke[J]. Expert Rev Neurother,2015,15(5):523-531.
[22] Xing J,Lu J. HIF-1alpha Activation Attenuates IL-6 and TNF-alpha Pathways in Hippocampus of Rats Following Transient Global Ischemia[J]. Cell Physiol Biochem,2016,39(2):511-520.
[23] Ziebell J M,Morganti-Kossmann M C. Involvement of pro-and anti-inflammatory cytokines and chemokines in the pathophysiology of traumatic brain injury[J]. Neurotherapeutics,2010,7(1):22-30.
[24] Vasconcelos J F,Teixeira M M,Barbosa-Filho J M,et al. The triterpenoid lupeol attenuates allergic airway inflammation in a murine model[J]. Int Immunopharmacol,2008,8(9):1216-1221.
[25] de Lima F O,Alves V,Barbosa Filho J M,et al. Antinociceptive effect of lupeol:evidence for a role of cytokines inhibition[J]. Phytother Res,2013,27(10):1557-1563.
[26] Zhu Y,Li X,Chen J,et al. The pentacyclic triterpene Lupeol switches M1 macrophages to M2 and ameliorates experimental inflammatory bowel disease[J]. Int Immunopharmacol,2016,30:74-84.
[27] Kim H C,Kim E,Bae J I,et al. Sevoflurane Postconditioning Reduces Apoptosis by Activating the JAK-STAT Pathway After Transient Global Cerebral Ischemia in Rats[J]. J Neurosurg Anesthesiol,2017,29(1):37-45.
[2] Fernndez M A,de las Heras B,Garcia M D,et al. New insights into the mechanism of action of the anti-inflammatory triterpene lupeol[J]. J Pharm Pharmacol,2001,53(11):1533-1539.
[3] Wang C P,Shi Y W,Tang M,et al. Isoquercetin Ameliorates Cerebral Impairment in Focal Ischemia Through Anti-Oxidative, Anti-Inflammatory, and Anti-Apoptotic Effects in Primary Culture of Rat Hippocampal Neurons and Hippocampal CA1 Region of Rats[J]. Mol Neurobiol,2017,54(3):2126-2142.
[4] Sakamula R,Thong-Asa W. Neuroprotective effect of pcoumaric acid in mice with cerebral ischemia reperfusion injuries[J]. Metab Brain Dis,2018,33(3):765-773.DOI:10. 1007/s11011-018-0185-7.
[5] Mukherjee P K,Ahamed K F,Kumar V,et al. Protective effect of biflavones from araucaria bidwillii hook in rat cerebral ischemia/reperfusion induced oxidative stress[J]. Behav Brain Res,2007,178(2):221-228.
[6] Nitta M,Azuma K,Hata K,et al. Systemic and local injections of lupeol inhibit tumor growth in a melanomabearing mouse model[J]. Biomed Rep,2013,1(4):641-645.
[7] Saleem M. Lupeol,a novel anti-inflammatory and anticancer dietary triterpene[J]. Cancer Lett,2009,285(2):109-115.
[8] 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.
[9]舒涛,刘畅,庞卯,等.丹酚酸B联合i PSCs-NSCs对大鼠脑缺血再灌注损伤后神经功能的修复作用及机制研究[J].中华神经医学杂志,2016,15(11):1084-1090.
[10] Kumari A,Kakkar P. Lupeol prevents acetaminophen-induced in vivo hepatotoxicity by altering the Bax/Bcl-2 and oxidative stress-mediated mitochondrial signaling cascade[J]. Life Sci,2012,90(15-16):561-570.
[11] Xiao A J,He L,Ouyang X,et al. Comparison of the anti-apoptotic effects of 15-and 35-minute suspended moxibustion after focal cerebral ischemia/reperfusion injury[J]. Neural Regen Res,2018,13(2):257-264. DOI:10. 4103/1673-5374. 226396.
[12]赵凯涛,杨婷,郝明华,等. miR-185与Apba-1在大鼠脑缺血再灌注损伤的表达变化及调控关系[J].解放军医药杂志,2016,28(3):13-17.
[13]杨红,郎晓芳,王勇.银杏叶提取物联合高压氧对局灶性脑缺血再灌注损伤大鼠外周血清氧化相关物质浓度的影响[J].解放军医药杂志,2015,27(10):34-38.
[14]谭永星,袁楠楠,夏裕宁,等. H2对大鼠全脑缺血再灌注损伤后海马CA1区神经元及树突棘的保护作用[J].中华神经医学杂志,2016,15(11):1091-1097.
[15] Sunitha S,Nagaraj M,Varalakshmi P. Hepatoprotective effect of lupeol and lupeol linoleate on tissue antioxidant defence system in cadmium-induced hepatotoxicity in rats[J]. Fitoterapia,2001,72(5):516-523.
[16] Prasad S,Kalra N,Shukla Y. Hepatoprotective effects of lupeol and mango pulp extract of carcinogen induced alteration in swiss albino mice[J]. Mol Nutr Food Res,2007,51(3):352-359.
[17] Yamashita K,Lu H,Lu J,et al. Effect of three triterpenoids,lupeol,betulin,and betulinic acid on the stimulus-induced superoxide generation and tyrosyl phosphorylation of proteins in human neutrophils[J]. Clin Chim Acta,2002,325(1-2):91-96.
[18] Prasad S,Kalra N,Singh M,et al. Protective effects of lupeol and mango extract against androgen induced oxidative stress in swiss albino mice[J]. Asian J Androl,2008,10(2):313-318.
[19] Sieber M W,Claus R A,Witte O W,et al. Attenuated inflammatory response in aged mice brains following stroke[J]. PLo S One,2011,6(10):e26288.
[20]孙恺,张秀萍,宋林杰,等.拮抗高迁移率族蛋白B1对糖尿病合并脑缺血再灌注损伤的保护作用研究[J].中华神经医学杂志,2016,15(9):901-907.
[21] Dziedzic T. Systemic inflammation as a therapeutic target in acute ischemic stroke[J]. Expert Rev Neurother,2015,15(5):523-531.
[22] Xing J,Lu J. HIF-1alpha Activation Attenuates IL-6 and TNF-alpha Pathways in Hippocampus of Rats Following Transient Global Ischemia[J]. Cell Physiol Biochem,2016,39(2):511-520.
[23] Ziebell J M,Morganti-Kossmann M C. Involvement of pro-and anti-inflammatory cytokines and chemokines in the pathophysiology of traumatic brain injury[J]. Neurotherapeutics,2010,7(1):22-30.
[24] Vasconcelos J F,Teixeira M M,Barbosa-Filho J M,et al. The triterpenoid lupeol attenuates allergic airway inflammation in a murine model[J]. Int Immunopharmacol,2008,8(9):1216-1221.
[25] de Lima F O,Alves V,Barbosa Filho J M,et al. Antinociceptive effect of lupeol:evidence for a role of cytokines inhibition[J]. Phytother Res,2013,27(10):1557-1563.
[26] Zhu Y,Li X,Chen J,et al. The pentacyclic triterpene Lupeol switches M1 macrophages to M2 and ameliorates experimental inflammatory bowel disease[J]. Int Immunopharmacol,2016,30:74-84.
[27] Kim H C,Kim E,Bae J I,et al. Sevoflurane Postconditioning Reduces Apoptosis by Activating the JAK-STAT Pathway After Transient Global Cerebral Ischemia in Rats[J]. J Neurosurg Anesthesiol,2017,29(1):37-45.