黄芪注射液通过调节JNK和NF-κB通路抑制视网膜神经节细胞凋亡
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摘要
目的:观察黄芪注射液对视神经牵拉伤大鼠视神经保护作用并探讨其作用机制。方法:将66只SPF级健康Wistar雄性大鼠按随机数字表法分为假手术组、视神经牵拉伤模型组及黄芪注射液治疗组3组各22只大鼠;横向定量牵拉法制作大鼠视神经损伤模型,假手术组仅暴露视神经不予牵拉;术后治疗组给予黄芪注射液腹腔注射,模型组给予生理盐水腹腔注射;治疗14 d后取大鼠视网膜组织,利用蛋白免疫印迹法检测JNK和NF-κB表达变化,并采用real time-PCR方法检测p75NTR的mRNA水平。结果:与假手术组比较,视神经牵拉伤模型大鼠视网膜组织p75NTR mRNA水平及JNK蛋白表达明显上升,NF-κB蛋白表达显著下降;给药14 d后与模型组比较,黄芪注射液治疗组大鼠视网膜组织p75NTR mRNA水平及JNK蛋白表达显著减少,NF-κB蛋白表达明显升高。结论:黄芪注射液具有视神经保护作用,其机制可能与抑制p75NTR mRNA的表达及JNK蛋白水平、促进NF-κB的蛋白表达有关。
Objective: To observe the optic nerve protective effects of astragalus membranaceus injection on tractive optic nerve injury in rats and to explore its possible mechanism. Methods: 66 healthy male Wistar rats were randomly divided into sham operated group,model group and astragalus membranaceus injection group,with 22 rats in each group;The optic nerve of model rats was injured by the transverse quantitative traction( tension given to the optic nerve: 0. 10N;tractive direction: perpendicular to the optic nerve; duration: 20 seconds),and that of sham-operation group was only exposed but not pulled; After model establishment,treatment group were given with astragalus membranaceus injection by intraperitoneal injection,and model group were given with normal saline for testing. 14 days later,the changes of expression of JNK and NF-κB in rat retinal tissue were detected with Western blotting,and the expression of p75 NTR mRNA was detected by real-time PCR. Results: Compared with the sham operation group,the expression level of p75 NTR mRNA and JNK protein in the optic nerve injury group increased. On the contrary,the protein expression level of NF-κB significantly declined; After treatment for 14 days,the expression of p75 NTR mRNA and JNK protein decreased in the astragalus injection treatment group compared with the model group. However,the protein expression level of NF-κB increased. Conclusion: Astragalus membranaceus injection could has the function of protecting the optic nerve. The mechanism may be related with decreasing p75 NTR mRNA and the protein expression of JNK,and increasing the protein expression of NF-κB.
Objective: To observe the optic nerve protective effects of astragalus membranaceus injection on tractive optic nerve injury in rats and to explore its possible mechanism. Methods: 66 healthy male Wistar rats were randomly divided into sham operated group,model group and astragalus membranaceus injection group,with 22 rats in each group;The optic nerve of model rats was injured by the transverse quantitative traction( tension given to the optic nerve: 0. 10N;tractive direction: perpendicular to the optic nerve; duration: 20 seconds),and that of sham-operation group was only exposed but not pulled; After model establishment,treatment group were given with astragalus membranaceus injection by intraperitoneal injection,and model group were given with normal saline for testing. 14 days later,the changes of expression of JNK and NF-κB in rat retinal tissue were detected with Western blotting,and the expression of p75 NTR mRNA was detected by real-time PCR. Results: Compared with the sham operation group,the expression level of p75 NTR mRNA and JNK protein in the optic nerve injury group increased. On the contrary,the protein expression level of NF-κB significantly declined; After treatment for 14 days,the expression of p75 NTR mRNA and JNK protein decreased in the astragalus injection treatment group compared with the model group. However,the protein expression level of NF-κB increased. Conclusion: Astragalus membranaceus injection could has the function of protecting the optic nerve. The mechanism may be related with decreasing p75 NTR mRNA and the protein expression of JNK,and increasing the protein expression of NF-κB.
引文
[1]廖良,孔莹莹,韦企平,等.黄芪注射液对体外培养视网膜神经节细胞的保护作用[J].眼科新进展,2010,12:1101-1104.
[2]Ken-ichiro M,Miho Y,Yoshitsugu I,et al.Neuroprotective Effect of Transcorneal Electrical Stimulation on the Acute Phase of Optic Nerve Injury[J].Invest Ophthalmol Vis Sci,2007,48:2356-2361.
[3]张圣海,吴继红,孙兴怀.一种改良的大鼠视神经损伤定量模型的建立[C].亚洲实验动物学会联合会(AFLAS)第三次会议暨中国实验动物学会(CALAS)第八届学术年会,2008:135.
[4]Pfaffl MW.A new mathematical model for relative quantification in real-time RT-PCR[J].Nucl Acids Res,2001,29:2002-2007.
[5]Anders Nykjaer,Thomas E Willnow,Claus Munck Petersen.p75NTR-live or let die[J].Current Opinion in Neurobiology,2005,15(1):49-57.
[6]Bradley R.Kraemer,John P.Snow,Peter Vollbrecht,et al.A Role for the p75 Neurotrophin Receptor in Axonal Degeneration and Apoptosis Induced by Oxidative Stress[J].The Journal of Biological Chemistry,2014,8(1):21205-21216.
[7]Longhi L,Perego C,Ortolano F,et al.Tumor necrosis factor in traumatic brain injury:effects of genetic deletion of p55 or p75receptor[J].J Cereb Blood Flow Me Tab,2013,33(8):1182-1189.
[8]Lebrun-Julien F,Morquette,Douillette A,et al.Inhibition of p75NTR in glia potentiates Trk A-mediated survival of injured retinal ganglion cells[J].Molecular and Cellular Neuroscience,2009,40(4):410-420.
[9]Dhanasekaran DN,Reddy EP.JNK signaling in apoptosis[J].Oncogene,2008,27(48):6245-6251.
[10]Eleanor T.Coffey.Nuclear and cytosolic JNK signaling in neurons[J].Nature Reviews Neuroscience,2014,15:285-299.
[11]Mattson MP,Culmsee C,Yu Z.Roles of nuclear factor kappa B in neuronal survival and plasticity[J].J Neurochem,2000,74(2):443-456.
[12]Christian Engelmann,Falk Weih,onny Haenold.Role of nuclear factor kappa B in central nervous system regeneration[J].Neural Regeneration Research,2014,9(7):707-711.
[13]中华人民共和国药典委员会.中国药典[S].2010年版.北京:中国医药科技出版社,2010:283.
[14]李淑芳.中药黄芪药理作用研究进展[J].湖北中医杂志,2013,35(6):73-75.
[2]Ken-ichiro M,Miho Y,Yoshitsugu I,et al.Neuroprotective Effect of Transcorneal Electrical Stimulation on the Acute Phase of Optic Nerve Injury[J].Invest Ophthalmol Vis Sci,2007,48:2356-2361.
[3]张圣海,吴继红,孙兴怀.一种改良的大鼠视神经损伤定量模型的建立[C].亚洲实验动物学会联合会(AFLAS)第三次会议暨中国实验动物学会(CALAS)第八届学术年会,2008:135.
[4]Pfaffl MW.A new mathematical model for relative quantification in real-time RT-PCR[J].Nucl Acids Res,2001,29:2002-2007.
[5]Anders Nykjaer,Thomas E Willnow,Claus Munck Petersen.p75NTR-live or let die[J].Current Opinion in Neurobiology,2005,15(1):49-57.
[6]Bradley R.Kraemer,John P.Snow,Peter Vollbrecht,et al.A Role for the p75 Neurotrophin Receptor in Axonal Degeneration and Apoptosis Induced by Oxidative Stress[J].The Journal of Biological Chemistry,2014,8(1):21205-21216.
[7]Longhi L,Perego C,Ortolano F,et al.Tumor necrosis factor in traumatic brain injury:effects of genetic deletion of p55 or p75receptor[J].J Cereb Blood Flow Me Tab,2013,33(8):1182-1189.
[8]Lebrun-Julien F,Morquette,Douillette A,et al.Inhibition of p75NTR in glia potentiates Trk A-mediated survival of injured retinal ganglion cells[J].Molecular and Cellular Neuroscience,2009,40(4):410-420.
[9]Dhanasekaran DN,Reddy EP.JNK signaling in apoptosis[J].Oncogene,2008,27(48):6245-6251.
[10]Eleanor T.Coffey.Nuclear and cytosolic JNK signaling in neurons[J].Nature Reviews Neuroscience,2014,15:285-299.
[11]Mattson MP,Culmsee C,Yu Z.Roles of nuclear factor kappa B in neuronal survival and plasticity[J].J Neurochem,2000,74(2):443-456.
[12]Christian Engelmann,Falk Weih,onny Haenold.Role of nuclear factor kappa B in central nervous system regeneration[J].Neural Regeneration Research,2014,9(7):707-711.
[13]中华人民共和国药典委员会.中国药典[S].2010年版.北京:中国医药科技出版社,2010:283.
[14]李淑芳.中药黄芪药理作用研究进展[J].湖北中医杂志,2013,35(6):73-75.