珠子参皂苷通过激活 PI3K/Akt通路对小鼠脑缺血再灌注损伤的保护作用研究
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摘要
目的:研究珠子参皂苷对小鼠脑缺血再灌注损伤的保护作用及可能机制。方法:雄性昆明小鼠随机分为假手术组、模型组、珠子参皂苷(100 mg/kg)、珠子参皂苷(200 mg/kg)和尼莫地平组(2 mg/kg)。给药组灌胃给予相应的药物,假手术组和模型组灌胃给予同体积0.5%羧甲基纤维素钠溶液,每天1次,连续7 d。以上各组给药7 d后制作小鼠大脑中动脉闭塞模型,假手术组切开缝合不进行动脉阻塞,在缺血2 h再灌注24 h后进行神经功能缺损评分、脑水肿、梗死面积和脑组织形态学分析,血液中ROS、LDH、SOD、GSH-Px、CAT和MDA的水平检测,Western blot检测脑组织中p-PI3K、PI3K、p-Akt、Akt、Bcl-2、Bax、Cytochrome C、cleaved-caspase-9和cleaved-caspase-3蛋白表达。结果:珠子参皂苷(100和200 mg/kg)和尼莫地平(2 mg/kg)可显著改善脑缺血再灌注损伤小鼠神经功能缺损评分、脑水肿、梗死面积和脑组织形态学,降低血清ROS、LDH和MDA水平,升高SOD、GSH-Px、CAT活性,上调脑组织中p-PI3K、p-Akt、Bcl-2蛋白表达和升高Bcl-2/Bax比率,下调Bax、Cytochrome C、cleaved-caspase-9和cleaved-caspase-3蛋白表达。结论:珠子参皂苷对小鼠缺血再灌注损伤具有较好的保护作用,其作用机制可能与其激活PI3K/Akt通路和抑制线粒体介导的凋亡通路有关。
AIM: To study the protective effect and possible mechanism of saponins from Rhizoma Panacis Majoris(SRPM) on cerebral ischemia-reperfusion(I/R) injury in mice. METHODS: Male Kunming mice were randomly divided into sham operation group, model group, SRPM(100 mg/kg), SRPM(200 mg/kg) and nimodipine group(2 mg/kg). The drug groups were given corresponding drugs by gavage, while the sham operation group and the model group were given 0.5% sodium carboxymethyl cellulose solution by gavage once a day for 7 consecutive days. After 7 days of administration, the middle cerebral artery occlusion model was established in mice in the sham operation group. After 2 hours of ischemia and 24 hours of reperfusion, the neurological deficit score, brain edema, infarct area and brain histomorphology were analyzed. The levels of ROS, LDH, SOD, GSH-Px, CAT and MDA in blood were detected. The protein levels of p-PI3 K, PI3 K, p-Akt, Akt Bcl-2, Bax, cytochrome C, cleaved-caspase-9 and cleaved-caspase-3 in brain tissue were detected by Western blot. RESULTS:SRPM(100 and 200 mg/kg) and nimodipine(2 mg/kg) could significantly improve neurological deficit score, brain edema, infarct area and brain histomorphology in cerebral I/R injury mice, reduce the levels of ROS, LDH and MDA, increase SOD, GSH-Px, CAT activities in serum, up-regulate the protein expressions of p-PI3 K, p-Akt, Bcl-2, and Bcl-2/Bax ratio, down-regulate the protein expressions of Bax, cytochrome C, cleaved-caspase-9 and cleaved-caspase-3. CONCLUSION: The protective effect of SRPM on I/R injury mice may be related to its activation of PI3 K/Akt pathway and inhibition of mitochondrial-mediated apoptotic pathway.
AIM: To study the protective effect and possible mechanism of saponins from Rhizoma Panacis Majoris(SRPM) on cerebral ischemia-reperfusion(I/R) injury in mice. METHODS: Male Kunming mice were randomly divided into sham operation group, model group, SRPM(100 mg/kg), SRPM(200 mg/kg) and nimodipine group(2 mg/kg). The drug groups were given corresponding drugs by gavage, while the sham operation group and the model group were given 0.5% sodium carboxymethyl cellulose solution by gavage once a day for 7 consecutive days. After 7 days of administration, the middle cerebral artery occlusion model was established in mice in the sham operation group. After 2 hours of ischemia and 24 hours of reperfusion, the neurological deficit score, brain edema, infarct area and brain histomorphology were analyzed. The levels of ROS, LDH, SOD, GSH-Px, CAT and MDA in blood were detected. The protein levels of p-PI3 K, PI3 K, p-Akt, Akt Bcl-2, Bax, cytochrome C, cleaved-caspase-9 and cleaved-caspase-3 in brain tissue were detected by Western blot. RESULTS:SRPM(100 and 200 mg/kg) and nimodipine(2 mg/kg) could significantly improve neurological deficit score, brain edema, infarct area and brain histomorphology in cerebral I/R injury mice, reduce the levels of ROS, LDH and MDA, increase SOD, GSH-Px, CAT activities in serum, up-regulate the protein expressions of p-PI3 K, p-Akt, Bcl-2, and Bcl-2/Bax ratio, down-regulate the protein expressions of Bax, cytochrome C, cleaved-caspase-9 and cleaved-caspase-3. CONCLUSION: The protective effect of SRPM on I/R injury mice may be related to its activation of PI3 K/Akt pathway and inhibition of mitochondrial-mediated apoptotic pathway.
引文
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[15] Tu L,Wang Y,Chen D,et al.Protective effects of Notoginsenoside R1 via regulation of the PI3K-Akt-mTOR/JNK pathway in neonatal cerebral hypoxic-ischemic brain injury[J].Neurochem Res,2018,43 (6):1210- 1226.
[16] Huang LF,Chen CW,Zhang X,et al.Neuroprotective effect of curcumin against cerebral ischemia-reperfusion via mediating autophagy and inflammation[J].J Mol Neurosci,2018,64 (1):129-139.
[17] Guo XD,Sun GL,Zhou TT,et al.Small molecule LX2343 ameliorates cognitive deficits in AD model mice by targeting both amyloid β production and clearance[J].Acta Pharmacol Sin,2016,37 (10):1281-1297.
[18] Pan YD,Wang N,Xia PP,et al.Inhibition of Rac1 ameliorates neuronal oxidative stress damage via reducing Bcl-2/Rac1 complex formation in mitochondria through PI3K/ Akt/mTOR pathway[J].Exp Neurol,2018,300:149-166.
[19] Chong SJ,Low IC,Pervaiz S.Mitochondrial ROS and involvement of Bcl-2 as a mitochondrial ROS regulator[J].Mitochondrion,2014,19 (Part A):39-48.
[20] Han B,Jiang P,Li ZX,et al.Coptisine-induced apoptosis in human colon cancer cells (HCT-116) is mediated by PI3K/Akt and mitochondrial-associated apoptotic pathway[J].Phytomedicine,2018,48:152-160.
[21] Youle RJ,Strasser A.The BCL-2 protein family:opposing activities that mediate cell death[J].Nat Rev Mol Cell Biol,2008,9 (1):47-59.
[22] Ryu DS,Kim SH,Kwon JH,et al.Orostachys japonicus induces apoptosis and cell cycle arrest through the mitochondria-dependent apoptotic pathway in AGS human gastric cancer cells[J].Int J Oncol,2014,45(1):459-469.
[2] 国家药典委员会.中国药典(一部)[M].北京:中国医药科技出版社,2015:271-272.
[3] Chan HH,Sun HD,Reddy MVB,et al.Potent α-glucosidase inhibitors from the roots of Panax japonicus C.A.Meyer var.major[J].Phytochemistry,2010,71(1112):1360-1364.
[4] 金家红,贺海波,石孟琼,等.珠子参总皂苷对小鼠局灶性脑缺血的保护作用[J].第三军医大学学报,2011,33 (24):2631-2633.
[5] Ou JJ,Kou L,Liang LY,et al.MiR-375 attenuates injury of cerebral ischemia/reperfusion via targetting Ctgf [J].Biosci Rep,2017,37(6):BSR20171242.
[6] El-Marasy SA,Abdel-Rahman RF,Abd-Elsalam RM.Neuroprotective effect of vildagliptin against cerebral ischemia in rats[J].Naunyn Schmiedebergs Arch Pharmacol,2018,391 (10):1133-1145.
[7] Wang Y,Tu L,Li YB,et al.Notoginsenoside R1 protects against neonatal cerebral hypoxic-ischemic injury through estrogen receptor-dependent activation of endoplasmic reticulum stress pathways[J].J Pharmacol Exp Ther,2016,357 (3):591-605.
[8] He HB,Li XM,Li DJ,et al.Saponins from Rhizoma Panacis Majoris attenuate myocardial ischemia/reperfusion injury via the activation of the Sirt1/FoxO1/Pgc-1α and Nrf2/ antioxidant defense pathways in rats[J].Pharmacognosy Magazine,2018,14 (56):297- 307.
[9] Huang SL,He HB,Zou K,et al.Protective effect of tomatine against hydrogen peroxide-induced neurotoxicity in neuroblastoma (SH-SY5Y) cells[J].J Pharm Pharm,2014,66 (6):844-854.
[10] Kanavaki A,Spengos K,Moraki M,et al.Serum levels of S100b and NSE proteins in patients with non-transfusion-dependent thalassemia as biomarkers of brain ischemia and cerebral vasculopathy[J].Int J Mol Sci,2017,18 (12):pii:E2724.
[11] Leonard SS,Harris GK,Shi X.Metal-induced oxidative stress and signal transduction[J].Free Radic Biol Med,2004,37 (12):1921-1942.
[12] 徐慧,张民远,戴勤学.miR-145通过调节SOD活性介导人参皂苷Rb1对脑缺血再灌注损伤大鼠的脑保护作用[J].中国临床药理学与治疗学,2018,23(1):1215-1220.
[13] Khwanraj K,Madlah S,Grataitong K,et al.Comparative mRNA expression of eEF1A isoforms and a PI3K/Akt/mTOR pathway in a cellular model of Parkinson's disease[J].Parkinsons Dis,2016:8716016.
[14] Chen A,Xiong LJ,Tong Y,et al.Neuroprotective effect of brain-derived neurotrophic factor mediated by autophagy through the PI3K/Akt/mTOR pathway[J].Mol Med Rep,2013,8 (4):1011-1016.
[15] Tu L,Wang Y,Chen D,et al.Protective effects of Notoginsenoside R1 via regulation of the PI3K-Akt-mTOR/JNK pathway in neonatal cerebral hypoxic-ischemic brain injury[J].Neurochem Res,2018,43 (6):1210- 1226.
[16] Huang LF,Chen CW,Zhang X,et al.Neuroprotective effect of curcumin against cerebral ischemia-reperfusion via mediating autophagy and inflammation[J].J Mol Neurosci,2018,64 (1):129-139.
[17] Guo XD,Sun GL,Zhou TT,et al.Small molecule LX2343 ameliorates cognitive deficits in AD model mice by targeting both amyloid β production and clearance[J].Acta Pharmacol Sin,2016,37 (10):1281-1297.
[18] Pan YD,Wang N,Xia PP,et al.Inhibition of Rac1 ameliorates neuronal oxidative stress damage via reducing Bcl-2/Rac1 complex formation in mitochondria through PI3K/ Akt/mTOR pathway[J].Exp Neurol,2018,300:149-166.
[19] Chong SJ,Low IC,Pervaiz S.Mitochondrial ROS and involvement of Bcl-2 as a mitochondrial ROS regulator[J].Mitochondrion,2014,19 (Part A):39-48.
[20] Han B,Jiang P,Li ZX,et al.Coptisine-induced apoptosis in human colon cancer cells (HCT-116) is mediated by PI3K/Akt and mitochondrial-associated apoptotic pathway[J].Phytomedicine,2018,48:152-160.
[21] Youle RJ,Strasser A.The BCL-2 protein family:opposing activities that mediate cell death[J].Nat Rev Mol Cell Biol,2008,9 (1):47-59.
[22] Ryu DS,Kim SH,Kwon JH,et al.Orostachys japonicus induces apoptosis and cell cycle arrest through the mitochondria-dependent apoptotic pathway in AGS human gastric cancer cells[J].Int J Oncol,2014,45(1):459-469.