摘要
目的:观察补肾活血方对兔退变椎间盘组织形态结构及Wnt3a、β-catenin蛋白表达的影响。方法:选取3月龄健康兔40只,随机分对照组,模型组,中药低、中、高剂量组,每组8只,除对照组外,其余兔均进行椎间盘退变造模手术。造模后3d开始给药,对照组及模型组每日以0.9%氯化钠溶液灌胃,中药低、中、高剂量组每日予补肾活血方灌胃,连续给药8周后处死取材,采用Masson染色、免疫组化染色等观察椎间盘组织形态学变化,并运用Western blot法检测Wnt3a、β-catenin蛋白表达差异。结果:Masson染色:与模型组比较,各药物组椎间盘组织中胶原纤维含量显著增多,髓核及纤维环组织的裂隙减少;免疫组化染色:中剂量组Wnt3a、β-catenin蛋白阳性表达率均显著下调(P<0.05),低剂量组与高剂量组Wnt3a蛋白阳性表达率显著下调(P<0.05);Western blot法:中、高剂量组椎间盘组织中Wnt3a及β-catenin蛋白的表达均较模型组显著下调(P<0.05),低剂量组β-catenin蛋白的表达较模型组显著下调(P<0.05)。结论:补肾活血方可能通过抑制Wnt/β-catenin信号通路的激活,一定程度上起到延缓椎间盘退变的作用。
Objective: To observe the effects of Bushen Huoxue Decoction on the morphological structure of defunct intervertebral disc and the expression of Wnt3 a and β-catenin. Methods: Forty healthy rabbits aged three months were selected and randomly divided into five groups, which were the control group, the model group, the low-dose group of traditional Chinese medicine, the medium-dose group and the high-dose group, with 8 rabbits in each group. In addition to the normal control group, all the other rabbits underwent disc degeneration surgery. Three days after the model was established, normal saline was given daily to the control group and the model group instead of traditional Chinese medicine. The low-dose, medium-dose and high-dose groups of traditional Chinese medicine were given daily to Bushen Huoxue Decoction for 8 weeks. After the intervention, samples were sacrificed, and the intervertebral disc histopathological changes were observed by Masson staining, immunohistochemical staining and other detection methods, and the protein expression differences of Wnt3 a and β-catenin were detected by Western blot. Results: Masson showed that: compared with the model group, the collagen fiber content in intervertebral disc tissue of each drug group was significantly increased, and the gap of nucleus pulposus and annulus fibrosus tissue was reduced;immunohistochemical staining: Wnt3 a and β-catenin were significantly down-regulated in the middle dose group(P<0.05), and the expression rate of Wnt3 a in the low-dose group and high-dose group was significantly decreased(P<0.05);Western blot showed that the expression of Wnt3 a and β-catenin protein in intervertebral disc tissue of middle and high dose groups were significantly lower than that of model group(P<0.05), while the expression of β-catenin protein in low-dose group was significantly lower than that in model group(P<0.05). Conclusion: Bushen Huoxue Decoction may delay the degeneration of intervertebral disc by inhibiting the activation of Wnt/β-catenin signaling pathway.
作者
李凯明
朱立国
李玲慧
张清
王尚全
展嘉文
陈明
梁龙
谢瑞
殷京
LI Kai-ming;ZHU Li-guo;LI Ling-hui;ZHANG Qing;WANG Shang-quan;ZHAN Jia-wen;CHEN Ming;LIANG Long;XIE Rui;YIN Jing(Department of Spinal Surgery,Wangjing Hospital of China Academy of Chinese Medical Sciences,Beijing 100102,China;Beijing Key Laboratory of Orthopedics of Traditional Chinese Medicine,Beijing 100102,China;Department of Orthopaedics,Wangjing Hospital of China Academy of Chinese Medical Sciences,Beijing 100102,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2020年第12期6001-6005,共5页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金项目(No.81674005,No.81904230,No.81804120,No.81302992)
中央本级重大增减支项目“名贵中药资源可持续利用能力建设项目”(No.2060302)
北京市自然科学基金项目(No.7164313)
中央级公益性科研院所基本科研业务费专项资助(No.ZZ10-015)
中国博士后科学基金(No.2017M611125,No.2016M591364)。
