针刺通过Wnt3a/β-catenin信号通路对颅脑损伤大鼠模型神经功能的改善
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摘要
目的分析针刺通过Wnt3a蛋白/细胞内β-连环蛋白(β-catenin)信号通路促进颅脑损伤大鼠模型神经功能改善的机制。方法选择SPF级SD雄性大鼠40只,喂养1 w后随机分为A组(不做处理)、B组(假手术,不造成脑组织损伤)、C组(建立颅脑损伤模型)、D组(建立颅脑损伤模型后第2天接受针刺治疗),观察各组造模后1、3、7、14 d改良神经功能评分(m Nss),以尼氏染色法观察脑组织形态学,应用实时荧光定量PCR法(Q-PCR)、Western blot法检测损伤侧皮层Wnt3a与β-catenin的mRNA、蛋白表达水平。结果造模后第1天、第3天C组与D组m Nss评分高于A组、B组,第7天、第14天C组、D组m Nss评分下降,D组第7天m Nss评分低于C组(P<0. 05),但第14天C组、D组m Nss评分比较差异无统计学意义(P>0. 05);造模第14天尼氏染色镜观察到D组尼氏小体数量较C组多; Q-PCR检测发现,造模第14天,4组Wnt3a mRNA、β-catenin mRNA表达量差异有统计学意义(P<0. 05),且D组Wnt3a mRNA、β-catenin mRNA表达量均高于C组(P<0. 05); Western blot检测发现,造模第14天D组Wnt3a、β-catenin蛋白表达量高于其他三组(P<0. 05)。结论针刺治疗可能通过Wnt3a/β-catenin信号通路,使Wnt3a、β-catenin表达上升,而显著改善颅脑损伤模型大鼠神经功能。
Objective To analyze the mechanism of acupuncture for improving the neurological function of rat model with craniocerebral injury through Wnt3 a protein/intracellular β-catenin signaling pathway. Methods Forty male SPF SD rats were randomly divided into group A( no treatment),group B( sham operation,no brain tissue damage),group C( model of craniocerebral injury)and group D( given acupuncture treatment on the 2 ndday after the establishment of the model of craniocerebral injury) after 1 week of feeding. Modified neurological function scores( m Nss) of each group on 1,3,7 d and 14 d after modeling were observed. The morphology of brain tissue was observed by Nissl staining. The mRNA and protein expression levels of Wnt3 a and β-catenin in injured cortex were detected by real-time quantitative PCR( Q-PCR) and Western blot. Results On the 1 stand 3 rdday after modeling,the m Nss scores of group C and group D were higher than those of group A and group B. The m Nss scores of group C and group D decreased on the 7 thand 14 thday,and the m Nss score of group D was lower than group C on the 7 thday( P<0. 05). There was no significant difference in the m Nss scores between group C and group D on the 14 thday( P>0. 05). On the 14 thday of modeling,Nissl staining showed that the number of Nissl bodies in group D was more than that in group C. Q-PCR found that on the 14 thday of modeling,there were significant quantity differences in the expression of Wnt3 a mRNA and β-catenin mRNA in the four groups( P<0. 05). The expression quantities of Wnt3 a mRNA and β-catenin mRNA in group D were higher than those in group C( P<0. 05). It was found out by western blot detection that on the 14 thday of modeling,the expression quantities of Wnt3 a and β-catenin in group D were higher than those in the other three groups( P<0. 05). Conclusion Acupuncture treatment may increase the expression of Wnt3 a and β-catenin through Wnt3 a/β-catenin signaling pathway,and significantly improve the neurological function of rats with craniocerebral injury.
Objective To analyze the mechanism of acupuncture for improving the neurological function of rat model with craniocerebral injury through Wnt3 a protein/intracellular β-catenin signaling pathway. Methods Forty male SPF SD rats were randomly divided into group A( no treatment),group B( sham operation,no brain tissue damage),group C( model of craniocerebral injury)and group D( given acupuncture treatment on the 2 ndday after the establishment of the model of craniocerebral injury) after 1 week of feeding. Modified neurological function scores( m Nss) of each group on 1,3,7 d and 14 d after modeling were observed. The morphology of brain tissue was observed by Nissl staining. The mRNA and protein expression levels of Wnt3 a and β-catenin in injured cortex were detected by real-time quantitative PCR( Q-PCR) and Western blot. Results On the 1 stand 3 rdday after modeling,the m Nss scores of group C and group D were higher than those of group A and group B. The m Nss scores of group C and group D decreased on the 7 thand 14 thday,and the m Nss score of group D was lower than group C on the 7 thday( P<0. 05). There was no significant difference in the m Nss scores between group C and group D on the 14 thday( P>0. 05). On the 14 thday of modeling,Nissl staining showed that the number of Nissl bodies in group D was more than that in group C. Q-PCR found that on the 14 thday of modeling,there were significant quantity differences in the expression of Wnt3 a mRNA and β-catenin mRNA in the four groups( P<0. 05). The expression quantities of Wnt3 a mRNA and β-catenin mRNA in group D were higher than those in group C( P<0. 05). It was found out by western blot detection that on the 14 thday of modeling,the expression quantities of Wnt3 a and β-catenin in group D were higher than those in the other three groups( P<0. 05). Conclusion Acupuncture treatment may increase the expression of Wnt3 a and β-catenin through Wnt3 a/β-catenin signaling pathway,and significantly improve the neurological function of rats with craniocerebral injury.
引文
[1]戴求福.针刺对创伤性颅脑损伤大鼠TLR2/4-NFκB信号通路的影响[D].广州:暨南大学,2016.
[2]郑亚萍,康红钰. Wnt/β-catenin信号通路在白芍总苷干预大鼠心肌肥厚中的作用[J].中成药,2018,40(1):194-197.
[3]焦红蕾.山奈酚通过Wnt3a/beta-catenin信号通路促进实验性脑缺血小鼠神经功能改善及神经再生[D].石家庄:河北医科大学,2016.
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[5]邓勇,王键,谭辉,等.脑络欣通对气虚血瘀型中脑动脉阻塞再灌注大鼠海马及额顶叶皮质Wnt3a、Wnt5a和β-Catenin表达的影响[J].安徽中医药大学学报,2017,36(3):59-63.
[6]王淑辉,王薇,张海廷,等. Wnt3a调控大鼠骨髓间充质干细胞向胆碱能神经元分化的实验研究[J].卒中与神经疾病,2016,23(5):309-314.
[7]杨先炯,毛新建,罗庆,等. Wnt3a/β-catenin信号通路调节模拟微重力诱导的骨髓间充质干细胞增殖抑制[J].空间科学学报,2016,36(1):63-69.
[8]朱将虎.经典Wnt/β-catenin信号通路在神经干细胞增殖和分化中的作用[J].重庆医学,2016,45(34):4858-4860.
[9] Itokazu T,Hayano Y,Takahashi R,et al. lnvolvement of Wnt/β-catenin signaling in the development of neuropathic pain[J]. Neurosci Res,2014,79(1):34-40.
[10]王菲菲,韩光,杜英杰,等. Wnt/β-catenin信号通路在高压氧治疗大鼠神经病理性痛中的作用[J].临床麻醉学杂志,2016,32(2):171-174.
[11]李佳诺,孙忠人,尹洪娜,等.针刺对颅脑损伤后相关因素影响的动物实验研究进展[J].针灸临床杂志,2017,33(6):74-77.
[12] Lei J,Gao G,Jiang J. Acute traumatic brain in jury:is current management evidence based? An empirica1 analysis of systematic reviews[J]. J Neurotrauma,2013,30(7):529-537.
[13]陈声鑫.针刺对TBI大鼠脑组织Wnt3aβ-catenin信号通路的影响[D].广州:暨南大学,2015.
[14]李洪涛,刘昊,杨方军,等.电针对膝关节骨性关节炎大鼠关节软骨中Wnt3a、β-catenin蛋白表达的影响[J].针灸临床杂志,2017,33(3):65-68.
[15] Zhu Y,Li C,Sun A,et al. Quantification of microRNA-210in the cerebrospinal fluid and serum:Implications for Alzheimer's disease[J]. Exp Ther Med,2015,9(3):1013-1017.
[16] Ring A,Kim YM,Kahn M. Wnt/catenin signaling in adult stem cell physiology and disease[J]. Stem Cell Rev,2014,10(4):512-525.
[2]郑亚萍,康红钰. Wnt/β-catenin信号通路在白芍总苷干预大鼠心肌肥厚中的作用[J].中成药,2018,40(1):194-197.
[3]焦红蕾.山奈酚通过Wnt3a/beta-catenin信号通路促进实验性脑缺血小鼠神经功能改善及神经再生[D].石家庄:河北医科大学,2016.
[4] Zhang YM,Zhang YQ,Cheng SB,et al. Effect of acupuncture on proliferation and differentiation of neural stem cells in brain tissues of rats with traumatic brain injury[J]. Chin J Integr Med,2013,19(2):132-136.
[5]邓勇,王键,谭辉,等.脑络欣通对气虚血瘀型中脑动脉阻塞再灌注大鼠海马及额顶叶皮质Wnt3a、Wnt5a和β-Catenin表达的影响[J].安徽中医药大学学报,2017,36(3):59-63.
[6]王淑辉,王薇,张海廷,等. Wnt3a调控大鼠骨髓间充质干细胞向胆碱能神经元分化的实验研究[J].卒中与神经疾病,2016,23(5):309-314.
[7]杨先炯,毛新建,罗庆,等. Wnt3a/β-catenin信号通路调节模拟微重力诱导的骨髓间充质干细胞增殖抑制[J].空间科学学报,2016,36(1):63-69.
[8]朱将虎.经典Wnt/β-catenin信号通路在神经干细胞增殖和分化中的作用[J].重庆医学,2016,45(34):4858-4860.
[9] Itokazu T,Hayano Y,Takahashi R,et al. lnvolvement of Wnt/β-catenin signaling in the development of neuropathic pain[J]. Neurosci Res,2014,79(1):34-40.
[10]王菲菲,韩光,杜英杰,等. Wnt/β-catenin信号通路在高压氧治疗大鼠神经病理性痛中的作用[J].临床麻醉学杂志,2016,32(2):171-174.
[11]李佳诺,孙忠人,尹洪娜,等.针刺对颅脑损伤后相关因素影响的动物实验研究进展[J].针灸临床杂志,2017,33(6):74-77.
[12] Lei J,Gao G,Jiang J. Acute traumatic brain in jury:is current management evidence based? An empirica1 analysis of systematic reviews[J]. J Neurotrauma,2013,30(7):529-537.
[13]陈声鑫.针刺对TBI大鼠脑组织Wnt3aβ-catenin信号通路的影响[D].广州:暨南大学,2015.
[14]李洪涛,刘昊,杨方军,等.电针对膝关节骨性关节炎大鼠关节软骨中Wnt3a、β-catenin蛋白表达的影响[J].针灸临床杂志,2017,33(3):65-68.
[15] Zhu Y,Li C,Sun A,et al. Quantification of microRNA-210in the cerebrospinal fluid and serum:Implications for Alzheimer's disease[J]. Exp Ther Med,2015,9(3):1013-1017.
[16] Ring A,Kim YM,Kahn M. Wnt/catenin signaling in adult stem cell physiology and disease[J]. Stem Cell Rev,2014,10(4):512-525.