牛膝总皂苷抑制椎间盘纤维环细胞焦亡的Wnt/β-catenin通路机制  

Mechanism of inhibiting intervertebral disc annulus fibrosus cell pyroptosis by achyranthes bidentata total saponins through regulating Wnt/β-catenin pathway

作　　者：宋佳明 张德生[1] SONG Jiaming;ZHANG Desheng(The Second Affiliated Hospital of Hunan University of Traditional Chinese Medicine,Changsha 410005,China)

机构地区：[1]湖南中医药大学第二附属医院,湖南长沙410005

出　　处：《陕西中医》2025年第3期325-329,共5页Shaanxi Journal of Traditional Chinese Medicine

基　　金：湖南省卫生健康委员会科研课题(D202304077262);湖南省中药管理局重点课题(A2024037)。

摘　　要：目的:基于Wnt/β-catenin通路探讨牛膝总皂苷对抑制椎间盘纤维环细胞焦亡的影响。方法:将105只SPF级SD大鼠随机分为牛膝总皂苷低、中、高剂量组,模型组,正常对照组,假手术组,阻断剂组,每组15只。除正常对照组外,其余各组均进行大鼠腰椎间盘退变模型造模,假手术组仅暴露椎间盘后缝合,不造成退变。术后3 d,牛膝总皂苷组分别按60、120、240μg/g剂量灌胃给药;阻断剂组皮下注射Wnt/β-catenin通路抑制剂;正常对照组、假手术组、模型组则给予等量0.9%氯化钠溶液灌胃。连续干预2周后,采用Western blot和qPCR技术检测各组椎间盘纤维环中Caspase-1、IL-18、IL-1β的蛋白和mRNA表达水平。结果:qPCR检测结果显示,与正常对照组、假手术组相比,模型组可见Caspase-1、IL-1β、IL-18 mRNA表达显著上调,经牛膝总皂苷药物干预后,与模型组相比较,牛膝总皂苷组Caspase-1、IL-1β、IL-18 mRNA表达水平显著降低,差异有统计学意义(均P<0.01)。经阻断剂药物干预后,与模型组相比较,Caspase-1、IL-1β、IL-18 mRNA表达显著降低(均P<0.01)。Western blot检测结果显示,与正常对照组、假手术组对照相比,模型组可见Caspase-1、IL-1β、IL-18蛋白表达水平上调,经牛膝总皂苷药物干预后,与模型组比较,牛膝总皂苷组Caspase-1、IL-1β、IL-18蛋白表达降低,差异有统计学意义(均P<0.01)。经阻断剂药物干预后,与模型组相比较,Caspase-1、IL-1β、IL-18蛋白表达显著降低(均P<0.01)。结论:牛膝总皂苷抑制椎间盘纤维环细胞焦亡的机制可能与调控Wnt/β-catenin信号通路有关。Objective:To investigate effect of total saponins of achyranin on the inhibition of focal death of intervertebral disc fibroring cells based on Wnt/β-catenin pathway.Methods:105 SPF-grade SD rats were randomly divided into low,medium,and high dose groups of total saponin of hyssop,model group,normal control group,sham-operation group,and blocker group,with 15 rats in each group.Except the normal control group underwent modelling of lumbar disc degeneration model in rats,and the sham-operation group only exposed the disc and then sutured it without causing degeneration.Starting from 3 days after surgery,the total saponin of hyssop group was administered by gavage at the doses of 60μg/g,120μg/g,240μg/g,respectively.The blocker group were injected subcutaneously with Wnt/β-catenin pathway inhibitor,and equal amounts of saline were given by gavage to the normal control group,the sham-operated group,and the modelling group.After 2 weeks of continuous intervention,the protein and mRNA expression levels of Caspase-1,IL-18,and IL-1β in the intervertebral disc fibrous annulus of each group were detected by Western blot and qPCR.Results:The results of qPCR assay showed that expression levels of Caspase-1,IL-1βand IL-18 mRNA were significantly up-regulated in the model group compared with the control of normal group and sham operation group(all P<0.01).After the drug intervention of total saponin of cow’s knee,the expression levels of Caspase-1,IL-1βand IL-18 mRNA in the group of total saponin of cow’s knee,compared with that in the model group,difference statistically significant(all P<0.01).After the blocker drug intervention,compared with the model group,expression levels of Caspase-1,IL-1β and IL-18 mRNA were significantly reduced(all P<0.01).The results of Western blot showed that,compared with the normal group and the sham-operated group control,the model group showed significant up-regulation of the expression levels of Caspase-1,IL-1βand IL-18 proteins,and protein expression levels were significantly u

关 键 词：牛膝总皂苷 WNT/Β-CATENIN 信号通路 椎间盘纤维环细胞 细胞焦亡 半胱氨酸蛋白酶-1 白细胞介素-18 白细胞介素-1Β 

分 类 号：R28[医药卫生—中药学]