芪芍通痹胶囊对腰椎间盘退行性改变模型大鼠Wnt/β-catenin通路的影响  

The Effect of Qishao Tongbi Capsule(芪芍通痹胶囊)on the Wnt/β-catenin Pathway in a Rat Model of Intervertebral Disc Degeneration

作　　者：薛玉满[1] 徐西林[2] 韩巍 许嘉犇[1] 许文婷 刘泽霖 张晓峰[3] XUE Yumen;XU Xilin;HAN Wei;XU Jiaben;XU Wenting;LIU Zelin;ZHANG Xiaofeng(The Second Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin,150000;The Third Affiliated Hospital of Heilongjiang University of Chinese Medicine;Heilongjiang University of Chinese Medicine)

机构地区：[1]黑龙江中医药大学附属第二医院,黑龙江哈尔滨150000 [2]黑龙江中医药大学附属第三医院 [3]黑龙江中医药大学

出　　处：《中医杂志》2025年第1期79-88,共10页Journal of Traditional Chinese Medicine

基　　金：全国名老中医药专家传承工作室建设项目(国中医药人教函[2022]75号);国家中医药管理局青年岐黄学者支持项目(2021-2023);黑龙江中医药大学科研基金项目(201730);黑龙江省第二批省级名中医师承项目(2020-2023)。

摘　　要：目的探讨芪芍通痹胶囊治疗腰椎间盘退行性改变(IDD)的可能作用机制。方法75只SD大鼠随机分为对照组,模型组,芪芍通痹低、高剂量组,芪芍通痹高剂量+激动剂组,每组15只。除对照组外,其余各组采用纤维环穿刺法制备大鼠IDD模型。造模后芪芍通痹低、高剂量组大鼠给予芪芍通痹胶囊0.2、0.8 g/(kg·d)灌胃;芪芍通痹高剂量+激动剂组给予芪芍通痹胶囊0.8 g/(kg·d)和SKL2001溶液10 mg/(kg·d)灌胃;对照组、模型组给予10 ml/(kg·d)蒸馏水灌胃,各组均每天1次,连续灌胃4周。最后一次给药后采用X射线和MRI检测各组IDD程度;HE染色、番红O-固绿染色观察椎间盘组织形态;免疫组化染色法观察椎间盘组织蛋白多糖、Ⅱ型胶原(COLⅡ)、基质金属蛋白酶3(MMP-3)蛋白水平;Western Blot法检测椎间盘髓核组织细胞外基质(ECM)相关蛋白[蛋白多糖、COLⅡ、MMP-3、基质金属蛋白酶9(MMP-9)、基质金属蛋白酶13(MMP-13)]、衰老相关蛋白(P53、P21、P16)、凋亡相关蛋白[B细胞淋巴瘤/白血病2(BCL-2)、BCL-2相关X蛋白(BAX)、剪切型天冬氨酸-半胱氨酸蛋白酶3(Cleaved Caspase-3)]、Wnt/β-catenin通路相关蛋白[Wnt3a、糖原合酶激酶-3β(GSK-3β)、β-catenin]水平;RT-qPCR检测椎间盘组织Wnt3a、GSK-3β、β-catenin mRNA表达。结果与模型组比较,芪芍通痹低、高剂量组大鼠椎间盘高度指数(DHI)升高,MRI Pfirrmann分级降低,IDD程度明显减轻,椎间盘髓核和纤维环结构完整性升高,髓核细胞数量增多,椎间盘髓核组织中蛋白多糖、COLⅡ、BCL-2、GSK-3β蛋白水平升高,MMP-3、MMP-9、MMP-13、P53、P21、P16、BAX、Cleaved Caspase-3、Wnt3a、β-catenin蛋白水平降低,Wnt3a、β-catenin mRNA表达降低,GSK-3βmRNA表达升高(P<0.05);与芪芍通痹低剂量组比较,芪芍通痹高剂量组大鼠DHI升高,MRI Pfirrmann分级降低(P<0.05),IDD程度进一步减轻,椎间盘髓核和纤维环结构完整性更高,髓核细胞数量进一步增�Objective To explore the possible mechanism of Qishao Tongbi Capsule(芪芍通痹胶囊,QTC)in the treatment of intervertebral disc degeneration(IDD).Methods Seventy-five rats were randomly divided into control group,model group,low-dose QTC group,high-dose QTC group,high-dose QTC+agonist group,with 15 rats in each group.Except for the control group,all other groups were subjected to a fibrous ring puncture to prepare an IDD model.After modeling,rats in low-dose QTC group and high-dose QTC group were given QTC at doses of 0.2 and 0.8 g/(kg·d)by gavage,respectively.Rats in high-dose QTC+agonist group was given QTC at 0.8 g/(kg·d)and SKL2001 solution at 10 mg/(kg·d)by gavage.The control group and model group were given 10 ml/(kg·d)distilled water by gavage.All treatments were given once a day for 4 consecutive weeks.After treatment,X-ray and magnetic resonance imaging(MRI)were used to detect IDD degree.Hematoxylin-eosin(HE)staining and Safranin O-Fast Green staining were used to observe the morphological changes of the intervertebral disc tissue.Immunohistochemical staining was performed to examine the levels of proteoglycan,typeⅡcollagen(COLⅡ),and matrix metalloproteinase-3(MMP-3)in the intervertebral disc tissue.Western blotting was used to detect the extracellular matrix(ECM)-related proteins(proteoglycan,COLⅡ,MMP-3,MMP-9,MMP-13),aging-related proteins(P53,P21,P16),apoptosis related proteins,including B-cell lymphoma/leukemia 2(BCL-2),BCL-2 related X protein(BAX),Cleaved Caspase-3,and Wnt/β-catenin pathway related proteins such as Wnt3a,glycogen synthase kinase-3β(GSK-3β)andβ-catenin in the intervertebral disc nucleus pulposus(NP)tissue.Reverse Transcription Quantitative Polymerase Chain Reaction(RT-qPCR)was used to assess the mRNA expression of Wnt3a,GSK-3β,andβ-catenin in intervertebral disc tissue.Results Compared with the model group,rats in the low-dose QTC group and high-dose QTC group exhibited improved DHI,decreased Pfirmann grading,and alleviated IDD.The structural integrity of the NP an

关 键 词：腰椎间盘退行性改变 髓核细胞 WNT/Β-CATENIN通路 芪芍通痹胶囊 

分 类 号：R285.5[医药卫生—中药学]