补肾益髓胶囊补泻成分对EAE小鼠大脑内Olig1、Olig2表达的作用研究
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摘要
目的观察梓醇配伍大黄素对实验性变态反应性脑脊髓炎(EAE)小鼠大脑内少突细胞转录因子(Olig) 1和Olig2表达的影响,探讨梓醇配伍大黄素促进髓鞘损伤修复的作用机制。方法 80只C57BL/6小鼠,随机分为正常组、模型组、激素组、配伍组(梓醇40 mg/kg、大黄素1 mg/kg),每组20只。于免疫当天,髓鞘少突胶质细胞糖蛋白35-55(MOG35-55)多肽皮下注射诱导产生EAE。于免疫诱导后第18天(症状高峰期)及第40天(症状缓解期)进行取材。透射电镜(TEM)观察髓鞘脱失情况,蛋白质免疫印迹法(WB)观察Olig1、Olig2蛋白的表达情况。实时荧光定量反转录聚合酶链式反应(Real-time rt-PCR)观察Olig1、Olig2基因表达情况。结果梓醇、大黄素合用能够显著减轻EAE小鼠髓鞘损伤,降低症状高峰期及症状缓解期神经功能评分。TEM显示,配伍组髓鞘环形层状结构较模型组小鼠破坏松散程度减轻。WB结果显示,第40天配伍组较模型组Olig2蛋白表达升高(P <0. 05),Olig1蛋白表达与模型组比较差异无统计学意义(P> 0. 05)。Real-time rt-PCR结果显示,第18天及第40天配伍组较模型组Olig1、Olig2基因表达均升高(P <0. 05)。结论梓醇、大黄素合用能够缓解EAE髓鞘损伤,增加EAE小鼠脑内Olig1、Olig2的表达,对髓鞘再生具有一定的促进作用。
Objective To observe the effects of catalpol and rheum emodin on the expressions of oligodendrocyte transcription factor-1(olig1) and olig2 in the cerebrum of mice with experimental autoimmune encephalomyelitis(EAE)and explore the effect mechanism of catalpol and rheum emodin on repairing the damaged myelin. Methods A total of 80 C57 BL/6 mice were randomized into a normal group,a model group,a hormone group and a drug composition group(catalpol 40 mg/kg,rheum emodin 1 mg/kg),20 mice in each one. On the day of immunization,EAE was induced by subcutaneous injection with myelin oligodendrocyte lucoprotein 35-55 IMOG 35-55 polypeptide. On day 18 after immune induction(symptom peak) and the day 40(symptom remission),the materials were collected. Using transmission electron microscope(TEM),myelinoclasis was observed. Using western blotting test,the expressions of olig1 and olig2 proteins were observed.The real-time reverse transcription-polymerase chain reaction(real-time RT-PCR) was used to observe the gene expressions of olig1 and olig2. Results The composition of catalpol and rheum emodin significantly alleviates myelin damage in EAE mice and reduces the nerve function scores in symptom peak and remission stages. TEM indicated that the destruction of myelin annular structure in the composition group was alleviated as compared with the model group. WB test showed that olig2 protein expression in the composition group was increased as compared with the model group on day 40(P <0. 05) and the difference was not significant in olig1 protein expression(P > 0. 05). Real-time RT-PCR results indicated that the gene expressions of olig1 and olig2 were increased in the composition group as compared with the model group on day 18 and 40(P < 0. 05). Conclusion The composition of catalpol and rheum emodin relieves myelin damage,increases the expressions of olig1 and olig2 in the brain of EAE mice. It promotes remyelination.
Objective To observe the effects of catalpol and rheum emodin on the expressions of oligodendrocyte transcription factor-1(olig1) and olig2 in the cerebrum of mice with experimental autoimmune encephalomyelitis(EAE)and explore the effect mechanism of catalpol and rheum emodin on repairing the damaged myelin. Methods A total of 80 C57 BL/6 mice were randomized into a normal group,a model group,a hormone group and a drug composition group(catalpol 40 mg/kg,rheum emodin 1 mg/kg),20 mice in each one. On the day of immunization,EAE was induced by subcutaneous injection with myelin oligodendrocyte lucoprotein 35-55 IMOG 35-55 polypeptide. On day 18 after immune induction(symptom peak) and the day 40(symptom remission),the materials were collected. Using transmission electron microscope(TEM),myelinoclasis was observed. Using western blotting test,the expressions of olig1 and olig2 proteins were observed.The real-time reverse transcription-polymerase chain reaction(real-time RT-PCR) was used to observe the gene expressions of olig1 and olig2. Results The composition of catalpol and rheum emodin significantly alleviates myelin damage in EAE mice and reduces the nerve function scores in symptom peak and remission stages. TEM indicated that the destruction of myelin annular structure in the composition group was alleviated as compared with the model group. WB test showed that olig2 protein expression in the composition group was increased as compared with the model group on day 40(P <0. 05) and the difference was not significant in olig1 protein expression(P > 0. 05). Real-time RT-PCR results indicated that the gene expressions of olig1 and olig2 were increased in the composition group as compared with the model group on day 18 and 40(P < 0. 05). Conclusion The composition of catalpol and rheum emodin relieves myelin damage,increases the expressions of olig1 and olig2 in the brain of EAE mice. It promotes remyelination.
引文
[1]胡学强,常艳宇.我国多发性硬化诊治现状与进展[J].重庆医科大学学报,2017,42(6):669-671.
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[9]Yang T,Zheng Q,Zhao H,et al.Effect of Bushen Yisui Capsule on oligodendrocyte lineage genes 1 and 2 in mice with experimental autoimmune encephalomyelitis[J].Chin J Integr Med,2016,22(12):932-940.
[10]Sherman DL,Brophy PJ.Mechanisms of axon ensheathment and myelin growth[J].Nat Rev Neurosci,2005,6(9):683-690.
[11]Li H,Richardson WD.Evolution of the CNS myelin gene regulatory program[J].Brain Res,2016,1641(Pt A):111-121.
[12]吕合作,邹健,王艳霞,等.Olig1过表达对大鼠神经干细胞向少突胶质细胞分化的影响[J].神经解剖学杂志,2010,26(2):201-205.
[13]Arnett HA,Fancy SP,Alberta JA,et al.bHLH transcription factor Olig1 is required to repair demyelinated lesions in the CNS[J].Science,2004,306(5704):2111-2115.
[14]Copray S,Balasubramaniyan V,Levenga J,et al.Olig2 overexpression induces the in vitro differentiation of neural stem cells into mature oligodendrocytes[J].Stem Cells,2006,24(4):1001-1010.
[15]Tan B,Yu J,Yin Y,Jet al.The Olig family affects central nervous system development and disease[J].Neural Regen Res,2014,9(3):329-36.
[2]樊永平,王苏.中医辨证治疗对复发缓解型多发性硬化患者复发率的影响[J].中医杂志,2015,56(8):683-685.
[3]樊永平.中医药辨证治疗多发性硬化的优势与不足[J].北京中医,2005(4):209-211.
[4]周莉,樊永平.二黄方减少多发性硬化轴索损伤和复发的临床研究[J].中国中医药信息杂志,2012,19(8):14-15,30.
[5]樊永平,王平,张星虎,等.二黄方治疗多发性硬化急性发作的临床观察[J].北京中医药大学学报,2006(4):273-276,280.
[6]房玲,樊永平,赵晖,等.补肾益髓方防治多发性硬化的研究进展[J].中医药导报,2013,19(12):108-110.
[7]Yang T,Zheng Q,Wang S,et al.Effect of catalpol on remyelination through experimental autoimmune encephalomyelitis acting to promote Olig1 and Olig2 expressions in mice[J].BMC Complement Altern Med,2017,17(1):240.
[8]李康宁,樊永平,王蕾.不同剂量MOG(35-55)抗原诱导EAE小鼠模型的免疫组织化学比较[J].中国实验动物学报,2010,18(6):451-456.
[9]Yang T,Zheng Q,Zhao H,et al.Effect of Bushen Yisui Capsule on oligodendrocyte lineage genes 1 and 2 in mice with experimental autoimmune encephalomyelitis[J].Chin J Integr Med,2016,22(12):932-940.
[10]Sherman DL,Brophy PJ.Mechanisms of axon ensheathment and myelin growth[J].Nat Rev Neurosci,2005,6(9):683-690.
[11]Li H,Richardson WD.Evolution of the CNS myelin gene regulatory program[J].Brain Res,2016,1641(Pt A):111-121.
[12]吕合作,邹健,王艳霞,等.Olig1过表达对大鼠神经干细胞向少突胶质细胞分化的影响[J].神经解剖学杂志,2010,26(2):201-205.
[13]Arnett HA,Fancy SP,Alberta JA,et al.bHLH transcription factor Olig1 is required to repair demyelinated lesions in the CNS[J].Science,2004,306(5704):2111-2115.
[14]Copray S,Balasubramaniyan V,Levenga J,et al.Olig2 overexpression induces the in vitro differentiation of neural stem cells into mature oligodendrocytes[J].Stem Cells,2006,24(4):1001-1010.
[15]Tan B,Yu J,Yin Y,Jet al.The Olig family affects central nervous system development and disease[J].Neural Regen Res,2014,9(3):329-36.