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

Improvement of the Neurological Function of Rat Model with Craniocerebral Injury by Acupuncture Through Wnt3a/β-catenin Signaling Pathway

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 Wnt3a protein/intracellularβ-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^nd day after the establishment of the model of craniocerebral injury)after 1 week of feeding.Modified neurological function scores(mNss)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 Wnt3a andβ-catenin in injured cortex were detected by real-time quantitative PCR(Q-PCR)and Western blot.Results On the 1^st and 3^rd day after modeling,the mNss scores of group C and group D were higher than those of group A and group B.The mNss scores of group C and group D decreased on the 7^th and 14^th day,and the mNss score of group D was lower than group C on the 7^th day(P<0.05).There was no significant difference in the mNss scores between group C and group D on the 14^th day(P>0.05).On the 14^th day 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^th day of modeling,there were significant quantity differences in the expression of Wnt3a mRNA andβ-catenin mRNA in the four groups(P<0.05).The expression quantities of Wnt3a mRNA andβ-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^th day of modeling,the expression quantities of Wnt3a andβ-catenin in group D were higher than those in the other three groups(P<0.05).Conclusion Acupuncture treatment may increase the expression of Wnt3a andβ-catenin through Wnt3a/β-catenin signaling pathway,and significantly improve the neurological function of rats with craniocerebral injury.