中医正骨2025，Vol.37Issue(3)：5-13,9.
补肾活血方对腰椎间盘退变小鼠腰椎终板软骨的影响及其作用机制
Effects and mechanism of Bushen Huoxue Fang(补肾活血方)on lumbar endplate cartilage in mice with inter-vertebral disc degeneration:an experimental study
摘要
Abstract
Objective:To observe the effects of Bushen Huoxue Fang(补肾活血方,BSHXF)on lumbar endplate cartilage in mice with intervertebral disc degeneration(IVDD),and to explore its underlying mechanism.Methods:Eighteen 10-week-old SPF-grade male C57BL/6J mice were randomized into blank control group,model group,and BSHXF group,6 cases in each group.All mice but the ones in blank control group were modeled by lumbar spine instability(LSI)surgery for inducing IVDD.The next day after successful modeling,the mice in BSHXF group were intervened by intragastric administration with BSHXF extract,while the ones in blank control group and model group with the same dose of normal saline,once a day for consecutive 8 weeks.After the end of the last intervention,all mice were sacrificed by cervical dislocation,and their L4 and L5 vertebrae were harvested for Micro-CT examination,and the relative height of the L4-5 intervertebral disc was calculated.Furthermore,the pathological changes of the lumbar endplate cartilage tissues were observed,and the morphological score of lumbar endplate cartilage tissues was calculated,meanwhile,the expression levels of collagen type Ⅱ alpha 1 chain(Col2α1),matrix metalloproteinase-13(MMP-13),and pSmad2 in the lumbar endplate cartilage tissues were detected.Results:①The Micro-CT analysis re-sults of the lumbar vertebrae.The blank control group and BSHXF group presented lower bone volume fraction(BVF)and fewer trabecular number(Tb.N)in the ossification area of lumbar endplate cartilage,and higher relative height of intervertebral disc compared to model group(P=0.000,P=0.022;P=0.002,P=0.001;P=0.000,P=0.000).Moreover,the BSHXF group exhibited higher BVF compared to blank control group(P=0.003),while,there was no significant differences between the 2 groups in the Tb.N and relative height of inter-vertebral disc(P=0.882,P=0.794).②The pathological observation of the lumbar endplate cartilage tissues.In blank control group,the lumbar endplate chondrocytes presented as low heterogeneity,normal cartilage thickness,and a small ossified area.Compared with that of blank control group,the changes in the lumbar endplate chondrocytes,manifesting as significantly increased heterogeneity,thinned cartilage layer,and an enlarged ossified area,were observed in mice of model group;while,compared with that of model group,the changes in the lumbar endplate chondrocytes,manifesting as reduced heterogeneity,increased cartilage thickness,and a decreased ossified area,were ob-served in mice of BSHXF group,but all these did not reach the levels of blank control group.Besides,the morphological score of lumbar endplate cartilage tissues was lower in blank control group and BSHXF group compared to model group(P=0.000,P=0.000),with no significant difference between blank control group and BSHXF group(P=0.591).③The expression levels of MMP-13 and Col2α1 in the lumbar endplate cartilage tissues.The proportion of MMP-13-positive cells was lower,while the ratio of Col2α1-positive expression area was higher in blank control group and BSHXF group compared to model group(P=0.002,P=0.010;P=0.000,P=0.026).Furthermore,the ratio of Col2α1-positive expression area was higher in blank control group compared to BSHXF group(P=0.000),while there was no sig-nificant difference in the proportion of MMP-13-positive cells between the 2 groups(P=0.226).④Thie expression level of pSmad2 in the lumbar endplate cartilage tissues.A high percentage of pSmad2-positive cells were observed in the lumbar endplate cartilage tissues of blank control group.The proportion of pSmad2-positive cells in the lumbar endplate cartilage tissues was higher in blank control group and BSHXF group compared to model group(P=0.014,P=0.040),with no significant difference between blank control group and BSHXF group(P=0.444).Conclusion:BSHXF can delay the pathological progression of lumbar IVDD in mice.It may work by down-regulating the expres-sion of MMP-13,and up-regulating the expression of Col2α 1 through promoting the phosphorylation of Smad2 in lumbar endplate chondro-cytes.关键词
椎间盘退化/腰椎/补肾活血方/终板软骨/小鼠/动物实验Key words
intervertebral disc degeneration/lumbar vertebrae/Bushen Huoxue Fang/endplate cartilage/mice/animal experimentation引用本文复制引用
张若谷,葛钦文,邹凯奥,厉驹,金红婷,童培建,王萍儿..补肾活血方对腰椎间盘退变小鼠腰椎终板软骨的影响及其作用机制[J].中医正骨,2025,37(3):5-13,9.基金项目
浙江省自然科学基金探索项目(LY23H270005) （LY23H270005）
