银杏叶提取物对辐射损伤小鼠脾脏氧化应激和蛋白激酶C活性的影响
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摘要
目的:探讨银杏叶提取物(GBE)对辐射小鼠脾脏细胞氧化损伤相关蛋白表达的影响,阐明其作用机制。方法:将60只ICR小鼠随机分为空白对照(NC)组、辐射对照(IC)组(4.0Gyγ线照射)、低剂量GBE (IC+GBEL)组(4.0Gyγ线+5.0mg·kg-1 GBE)、中剂量GBE (IC+GBEM)组(4.0Gyγ线+10.0mg·kg-1 GBE)和高剂量GBE (IC+GBEH)组(4.0Gyγ线+20.0mg·kg-1 GBE),每组12只。各剂量GBE组小鼠连续给予相应剂量GBE共14d,第15天除NC组小鼠外均给予总剂量为4.0Gy的γ射线全身照射。于照射后24h采用生化方法检测各组小鼠脾组织中丙二醛(MDA)水平,超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-px)活性;免疫组织化学法观察各组小鼠脾脏细胞中8-羟基脱氧鸟嘌呤(8-OHdG)表达水平,ELISA法检测各组小鼠脾组织中蛋白激酶C (PKC)活性。结果:与NC组比较,IC组和各剂量GBE组小鼠脾组织中MDA水平和8-OHdG表达水平明显升高(P<0.05),SOD、CAT、GSH-px和PKC活性明显降低(P<0.05);与IC组比较,各剂量GBE组小鼠脾组织中MDA水平和8-OHdG表达水平明显降低(P<0.05),SOD、CAT、GSH-px和PKC活性明显升高(P<0.05)。结论:GBE可以抑制辐射损伤小鼠脾脏内的氧化应激,调节恢复脾脏PKC活性,从而发挥其对辐射损伤的保护作用。
Objective:To investigate the influence of Ginkgo biloba L.extract(GBE)in the expressions of oxidative damage-related proteins in the spleen tissue of the mice with radiation damage,and to clarify its mechanism.Methods:Sixty ICR mice were randomly divided into normal control(NC)group,irradiation control(IC)group(4.0 Gyγ-ray radiation),low dose of GBE(IC+GBEL)group(4.0 Gyγ-ray+5.0 mg·kg-1 GBE),medium dose of GBE(IC+GBEM)group(4.0 Gyγ-ray+10.0 mg·kg-1 GBE)and high dose of GBE(IC+GBEH)group(4.0 Gyγ-ray+20.0 mg·kg-1 GBE)(n=12).After continuous administration of GBE for 14 d,all mice received the whole body radiation ofγ-ray with the total dose of 4.0 Gy on the 15 th day except for the mice in NC group.The levels of malonaldehyde(MDA),the activities of superoxide dismutase(SOD),catalase(CAT)and glutathione peroxidase(GSH-px)in the spleen tissue of the mice in various groups were measured by biochemical method 24 hafter radiation.The expression levels of 8-hydroxy-2 deoxyguanosine(8-OHdG)in the spleen tissue of the mice in various groups were examined by immunohistochemistry.The activities of protein kinase C(PKC)in the spleen tissue of the mice in various groups were detected by ELISA.Results:Compared with NC group,the levels of MDA and 8-OHdG in the spleen tissue of the mice in IC and different doses of GBE groups were significantly increased(P<0.05),and the activities of SOD,CAT,GSH-px and PKC were significantly decreased(P<0.05).Compared with IC group,the levels of MDA and 8-OHdG in the spleen tissue of the mice in different doses of GBE groups were significantly decreased(P<0.05),and the activities of SOD,CAT,GSH-px and PKC were significantly increased(P<0.05).Conclusion:GBE can inhibit the oxidative stress and regulate the PKC activity in the spleen tissue of the mice with irradiation damage to play aprotective role against the radiation damage.
Objective:To investigate the influence of Ginkgo biloba L.extract(GBE)in the expressions of oxidative damage-related proteins in the spleen tissue of the mice with radiation damage,and to clarify its mechanism.Methods:Sixty ICR mice were randomly divided into normal control(NC)group,irradiation control(IC)group(4.0 Gyγ-ray radiation),low dose of GBE(IC+GBEL)group(4.0 Gyγ-ray+5.0 mg·kg-1 GBE),medium dose of GBE(IC+GBEM)group(4.0 Gyγ-ray+10.0 mg·kg-1 GBE)and high dose of GBE(IC+GBEH)group(4.0 Gyγ-ray+20.0 mg·kg-1 GBE)(n=12).After continuous administration of GBE for 14 d,all mice received the whole body radiation ofγ-ray with the total dose of 4.0 Gy on the 15 th day except for the mice in NC group.The levels of malonaldehyde(MDA),the activities of superoxide dismutase(SOD),catalase(CAT)and glutathione peroxidase(GSH-px)in the spleen tissue of the mice in various groups were measured by biochemical method 24 hafter radiation.The expression levels of 8-hydroxy-2 deoxyguanosine(8-OHdG)in the spleen tissue of the mice in various groups were examined by immunohistochemistry.The activities of protein kinase C(PKC)in the spleen tissue of the mice in various groups were detected by ELISA.Results:Compared with NC group,the levels of MDA and 8-OHdG in the spleen tissue of the mice in IC and different doses of GBE groups were significantly increased(P<0.05),and the activities of SOD,CAT,GSH-px and PKC were significantly decreased(P<0.05).Compared with IC group,the levels of MDA and 8-OHdG in the spleen tissue of the mice in different doses of GBE groups were significantly decreased(P<0.05),and the activities of SOD,CAT,GSH-px and PKC were significantly increased(P<0.05).Conclusion:GBE can inhibit the oxidative stress and regulate the PKC activity in the spleen tissue of the mice with irradiation damage to play aprotective role against the radiation damage.
引文
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[20]HUANG S X,QIU G Z,CHENG F R,et al.Berberine protects secondary injury in mice with traumatic brain injury through anti-oxidative and anti-inflammatory modulation[J].Neurochem Res,2018,43(9):1814-1825.
[21]刘勇,黄蔚初,程绪浩,等.电磁辐射对家兔小脑蛋白激酶C活力的影响[J].环境与健康杂志,2008,25(9):781-783.
[22]刘春明,李德新,吕爱萍,等.脾阳虚大鼠肝脾蛋白激酶C活性的实验研究[J].辽宁中医杂志,2000,27(1):44-45.
[23]赵忆宁,何颖,沈先荣,等.西咪替丁对低剂量电离辐射致大鼠氧化应激的保护作用[J].解放军医学杂志,2017,42(2):128-133.
[24]苌雪,王坤,张健,等.高能光子水吸收剂量辐射质转换因子的实验测量[J].中国医学物理学杂志,2017,34(12):1196-1199.
[2]PURBEY P K,SCUMPIA P O,KIM P J,et al.Defined sensing mechanisms and signaling pathways contribute to the global inflammatory gene expression output elicited by ionizing radiation[J].Immunity,2017,47(3):421-434.
[3]GAJOWIK A,DOBRZYNSKA M M.The evaluation of protective effect of lycopene against genotoxic influence of X-irradiation in human blood lymphocytes[J].Radiat Environ Biophys,2017,56(4):413-422.
[4]韩晓亮,蒲国华,李宝永,等.丙泊酚对电离辐射引起小鼠造血系统损伤的防护作用[J].国际药学研究杂志,2016,43(5):931-934.
[5]杨龙,王奕萍,荣曙,等.电离辐射对大鼠脑皮质牛磺酸、谷氨酸及其谷氨酸阳性神经元的影响[J].中国辐射卫生,2006,15(4):402-404.
[6]宋军,朱燕梅,王秋林.银杏叶提取物对小鼠心肌细胞氧化应激损伤的影响[J].成都医学院学报,2017,12(3):260-264.
[7]王冠,耿丽晶,陈博,等.不同前处理方式对银杏叶中过氧化氢酶活性的影响[J].饲料研究,2016(18):34-38.
[8]WANG Z,ZHANG J L,REN T K,et al.Targeted metabolomic profiling of cardioprotective effect of Ginkgo biloba L.extract on myocardial ischemia in rats[J].Phytomedicine,2016,23(6):621-631.
[9]宋媛媛,吕欣,高春时,等.银杏叶提取物对电离辐射下大鼠骨髓间充质干细胞的保护作用机制研究[J].中国实验诊断学,2016,20(8):1235-1239.
[10]陈家柏,伍炜培.银杏叶提取物与茶多酚对辐射损伤的协同保护作用[J].中药材,2004,27(9):664-665.
[11]金虹,黄毅,王继生,等.银杏叶提取物对辐射损伤小鼠的保护作用[J].中草药,2010,41(8):1339-1342.
[12]方连英,王彦,徐畅,等.电离辐射诱导造血干细胞损伤的分子机制研究进展[J].基础医学与临床,2017,37(2):256-260.
[13]MULINACCI N,VALLETTA A,PASQUALETTI V,et al.Effects of ionizing radiation on bio-active plant extracts useful for preventing oxidative damages[J].Nat Prod Res,2018:1-9.
[14]SUMAN S,KUMAR S,FORNACE A J,et al.The effect of carbon irradiation is associated with greater oxidative stress in mouse intestine and colon relative toγ-rays[J].Free Radic Res,2018,16:1-11.
[15]袁茵,王辉,鲁欣.银杏叶提取物对叔丁基过氧化氢损伤人脐带间充质干细胞的干预作用[J].广州中医药大学学报,2015,32(3):458-463.
[16]王如科,孙源源,韩建一,等.银杏叶提取物对过氧化氢刺激下的血管内皮细胞的影响[J].现代生物医学进展,2014,14(26):5038-5041,5048.
[17]ZHAO M,WANG X X,WAN W H.Effects of the ginkgo biloba extract on the superoxide dismutase activity and apoptosis of endothelial progenitor cells from diabetic peripheral blood[J].Genet Mol Res,2014,13(1):220-227.
[18]WANG A,YANG Q W,LI Q J,et al.Ginkgo biloba L.extract reduces H2O2-induced bone marrow mesenchymal stem cells cytotoxicity by regulating mitogen-activated protein kinase(MAPK)signaling pathways and oxidative stress[J].Med Sci Monit,2018,24:3159-3167.
[19]王俊妹,孙千月,吴哲,等.银杏叶提取物对破骨细胞分化和骨吸收的作用及其机制[J].吉林大学学报:医学版,2017,43(6):1130-1136.
[20]HUANG S X,QIU G Z,CHENG F R,et al.Berberine protects secondary injury in mice with traumatic brain injury through anti-oxidative and anti-inflammatory modulation[J].Neurochem Res,2018,43(9):1814-1825.
[21]刘勇,黄蔚初,程绪浩,等.电磁辐射对家兔小脑蛋白激酶C活力的影响[J].环境与健康杂志,2008,25(9):781-783.
[22]刘春明,李德新,吕爱萍,等.脾阳虚大鼠肝脾蛋白激酶C活性的实验研究[J].辽宁中医杂志,2000,27(1):44-45.
[23]赵忆宁,何颖,沈先荣,等.西咪替丁对低剂量电离辐射致大鼠氧化应激的保护作用[J].解放军医学杂志,2017,42(2):128-133.
[24]苌雪,王坤,张健,等.高能光子水吸收剂量辐射质转换因子的实验测量[J].中国医学物理学杂志,2017,34(12):1196-1199.