Experimental study on the regulation of Bushen Yisui formula and its disassembled formulas on related molecules of inhibitory signal pathway of axon regeneration in mice with experimental autoimmune encephalomyelitis

doi:10.3969/j.issn.1006-7795.2020.02.010

Abstract

Abstract: Objectives To observe the regulation of Bushen Yisui formula (BSYSF) and its disassembled formulas, Bushen formula (BSF) and Huatan Huoxue formula (HTHXF) on related inhibition molecules in signal pathway of axon regeneration in mice with experimental autoimmune encephalomyelitis (EAE). Methods Female C57BL/6 mice were randomly divided into normal control (NC), model (MO), prednisone acetate (PA, 6mg/kg), catapol (CA, 40 mg/kg), BSYSF (raw drug 3.02 g/kg), BSF (raw drug 1.44 g/kg) and HTHXF(raw drug 1.57 g/kg), with 16 mice in each group. EAE model in mice was made by immunizing with myelin oligodendrocyte glycoprotein _35-55 (MOG_35-55). The mice in NC and MO groups were given distilled water by means of intragastric administration, the mice in treatment groups were given corresponding drugs once a day for 40 d. The brain and spinal cord of mice were taken on day 25 and 40 of immunization. The gene expression of NogoA, NgR, RhoA and ROCKⅡmRNA were determined by quantitative real time-PCR, the protein expression of the above indexes were detected by the immunohistochemistry and Western blotting technique. Results The mRNA and protein expressions of NogoA, NgR, RhoA and ROCKⅡin the brain and spinal cord in MO mice were increased significantly in EAE mice compared to NC mice (P<0.05, P<0.01, P<0.001). After the treatment of BSYSF and its disassembled formulas BSF和HTHXF, the mRNA and protein expressions of NogoA, NgR, RhoA and ROCKⅡwere reduced to some extent, the above indexes had statistically significant in BSYSF group compared with MO group (P<0.05, P<0.01, P<0.001). Conclusions BSYSF and its disassembled formulas BSF and HTHXF had the regulation on inhibitory factors of axon regeneration NogoA/NgR and its signal pathway RhoA/ROCK in EAE mice, and BSYSF had obvious effect. These findings provide an experimental basis for explaining the mechanism of BSYSF promoting axon regeneration.

Key words: Bushen Yisui formula and its disassembled formulas, experimental autoimmune encephalomyelitis, axon regeneration, NogoA/NgR/RhoA/ROCK

CLC Number:

Wang Lei, An Chen, Zhao Hui, Xue Bing, Qi Fang, Li Junling, Jin Liangyun, Zhang Nan, Fan Yongping. Experimental study on the regulation of Bushen Yisui formula and its disassembled formulas on related molecules of inhibitory signal pathway of axon regeneration in mice with experimental autoimmune encephalomyelitis[J]. Journal of Capital Medical University, 2020, 41(2): 200-211.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2020.02.010

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2020/V41/I2/200

References

[1] Gajofatto A, Benedetti M D. Treatment strategies for multiple sclerosis:when to start, when to change, when to stop?[J]. World J Clin Cases, 2015, 3(7):545-555.
[2] Van Schependom J, Guldolf K, D'hooghe M B, et al. Correction to:detecting neurodegenerative pathology in multiple sclerosis before irreversible brain tissue loss sets in[J]. Transl Neurodegener, 2020, 9:3.
[3] Schultz V, van der Meer F, Wrzos C, et al. Acutely damaged axons are remyelinated in multiple sclerosis and experimental models of demyelination[J]. Glia, 2017, 65(8):1350-1360.
[4] Wootla B, Denic A, Watzlawik J O, et al. Antibody-mediated oligodendrocyte remyelination promotes axon health in progressive demyelinating disease[J]. Mol Neurobiol, 2016, 53(8):5217-5228.
[5] 中国免疫学会神经免疫分会,中华医学会神经病学分会神经免疫学组.多发性硬化诊断和治疗中国专家共识(2018版)[J].中国神经免疫学和神经病学杂志,2018,25(6):387-394.
[6] 陈克龙,樊永平.补肾化痰活血法对多发性硬化患者生存质量的影响[J].中华中医药学刊,2016,34(9):2141-2144.
[7] 樊永平,陈克龙,尤昱中,等.补肾益髓胶囊治疗肝肾阴虚复发缓解型多发性硬化临床疗效观察[J].中华中医药杂志,2018,33(9):4220-4222.
[8] 北京中医药学会脑病专业委员会.多发性硬化/视神经脊髓炎中医临床诊疗规范[J].首都医科大学学报,2018,39(6):833-835.
[9] 王永强,师一民,安辰,等.补肾益髓方促进实验性自身免疫性脑脊髓炎小鼠髓鞘再生的作用[J].环球中医药,2016,9(12):1443-1449.
[10] 孙豪,郑宏,计婧,等.补肾益髓方促进实验性自身免疫性脑脊髓炎小鼠小胶质细胞向M2极化的作用[J].首都医科大学学报,2018,39(6):798-804.
[11] 安辰,王永强,师一民,等.补肾益髓方及其拆方对实验性自身免疫性脑脊髓炎小鼠的神经保护作用及NF200和CSPGs表达的影响[J].中华中医药杂志,2016,31(4):1215-1220.
[12] 安辰,王永强,师一民,等.补肾益髓方及其拆方对实验性自身免疫性脑脊髓炎小鼠脑和脊髓中β-APP和MAP-2表达的影响[J].中国中医急症,2016,25(4):565-568,587.
[13] Kim M J, Kang J H, Theotokis P, et al. Can we design a Nogo receptor-dependent cellular therapy to target MS?[J].Cells, 2019, 8(1). pii:E1.
[14] Sandvig A, Berry M, Barrett L B, et al. Myelin-, reactive glia-, and scar-derived CNS axon growth inhibitors:expression, receptor signaling, and correlation with axon regeneration[J]. Glia, 2004, 46(3):225-251.
[15] Theotokis P, Lourbopoulos A, Touloumi O, et al. Time course and spatial profile of Nogo-A expression in experimental autoimmune encephalomyelitis in C57BL/6 mice[J]. J Neuropathol Exp Neurol, 2012, 71(10):907-920.
[16] Josephson A, Trifunovski A, Widmer H R, et al. Nogo-receptor gene activity:cellular localization and developmental regulation of mRNA in mice and humans[J]. J Comp Neurol, 2002, 453(3):292-304.
[17] Wang K C, Kim J A, Sivasankaran R, et al. P75 interacts with the Nogo receptor as a co-receptor for Nogo, MAG and OMgp[J]. Nature, 2002, 420(6911):74-78.
[18] Nakagawa O, Fujisawa K, Ishizaki T, et al. ROCK-Ⅰand ROCK-Ⅱ, two isoforms of Rho-associated coiled-coil forming protein serine/threonine Kinase in mice[J]. FEBS Lett, 1996, 392(2):189-193.
[19] Yan Y, Yu J, Gao Y, et al. Therapeutic potentials of the Rho kinase inhibitor Fasudil in experimental autoimmune encephalomyelitis and the related mechanisms[J]. Metab Brain Dis, 2019, 34(2):377-384.
[20] 刘蕾,郑琦,安辰,等.补肾配伍化痰活血药对低血清培养SH-SY5Y神经细胞轴突生长的影响[J].中医药信息,2014,31(5):5-9.
[21] Yuan C X, Chu T, Liu L, et al. Catalpol induces oligodendrocyte precursor cell-mediated remyelination in vitro[J]. Am J Transl Res, 2015, 7(11):2474-2481.
[22] 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.
[23] Satoh J, Onoue H, Arima K, et al. Nogo-A and nogo receptor expression in demyelinating lesions of multiple sclerosis[J]. J Neuropathol Exp Neurol, 2005, 64(2):129-138.
[24] Karnezis T, Mandemakers W, McQualter J L, et al. The neurite outgrowth inhibitor NogoA is involved in autoimmune-mediated demyelination[J]. Nat Neurosci, 2004, 7(7):736-744.
[25] Petratos S, Ozturk E, Azari M F, et al. Limiting multiple sclerosis related axonopathy by blocking Nogo receptor and CRMP-2 phosphorylation[J]. Brain, 2012, 135(Pt6):1794-1818.
[26] Fujita Y, Yamashita T. Axon growth inhibition by RhoA/ROCK in the central nervous system[J]. Front Neurosci, 2014, 8:338.
[27] Lingor P, Teusch N, Schwarz K, et al. Inhibition of Rho kinase (ROCK) increases neurite outgrowth on chondroitin sulphate proteoglycan in vitro and axonal regeneration in the adult optic nerve in vivo[J]. J Neurochem, 2007, 103(1):181-189.