摘要
目的探讨人参二醇型皂苷(PDS)对小鼠膝关节软骨组织损伤的修复作用及其机制。方法小鼠分为假手术组(Sham组,仅膝关节切开处理)、模型组(Model组,构建膝关节软骨损伤模型)、低剂量PDS组(L-PDS组,构建膝关节软骨损伤模型后2 mg/kg的PDS处理)和高剂量PDS组(H-PDS组,构建膝关节软骨损伤模型后10 mg/kg的PDS处理)。免疫组织化学法观察4组小鼠组织学;脱氧核苷酸末端转移酶介导的dUTP缺口末端标记(TUNEL)法检测细胞凋亡;酶联免疫吸附法检测基质金属蛋白酶-3(MMP-3)、转化生长因子-β1(TGF-β1)和肿瘤坏死因子(TNF-α)水平;实时荧光定量聚合酶链反应(qRT-PCR)法检测基质金属蛋白酶-13(MMP-13)和Ⅱ型胶原蛋白(COL-Ⅱ)基因信使核糖核酸(Messenger ribonucleic acid,mRNA)表达。两组间比较行t检验,多组间比较采用单因素方差分析(组间两两比较采用LSD-t检验)。结果Model组小鼠Mankin评分、凋亡率、MMP-3、TGF-β1和TNF-α水平、MMP-13基因mRNA表达显著高于Sham组小鼠[Mankin评分8.93±1.05比1.05±0.08,凋亡率(58.28±4.24)%比(4.48±0.87)%,MMP-3为(33.04±3.21)pg/ml比(8.52±1.26)pg/ml,TGF-β1为(5.09±0.68)pg/ml比(2.27±0.31)pg/ml,TNF-α分别为(14.92±2.31)pg/ml比(5.11±1.08)pg/ml,MMP-13基因mRNA表达为4.68±0.51比1.03±0.05],COL-Ⅱ基因mRNA表达显著低于Sham组小鼠(0.17±0.02比1.07±0.03),差异有统计学意义(t=19.478、29.126、11.375、15.128、9.233、8.306、11.233,P<0.05);L-PDS组小鼠Mankin评分、凋亡率、MMP-3、TGF-β1和TNF-α水平、MMP-13基因mRNA表达显著高于H-PDS组小鼠[Mankin评分(6.01±0.79)分比(3.54±0.68)分,凋亡率(36.67±3.45)%比(22.89±2.26)%,MMP-3为(21.09±2.18)pg/ml比(13.38±1.42)pg/ml,TGF-β1为(3.94±0.32)pg/ml比(2.98±0.29)pg/ml,TNF-α分别为(11.35±1.68)pg/ml比(8.33±1.02)pg/ml,MMP-13基因mRNA表达为2.17±0.36比1.46±0.21],COL-Ⅱ基因mRNA表达显著低于H-PDS组小鼠(0.54±0.08比0.77±0.11),差异有统计学意义(t=14.233、16.345
Objective To study the repairing effect of panaxadiol saponin(PDS)on the knee cartilage injury in mice and its mechanism.Methods Mice were divided into Sham group(only knee joint incision treatment),model group(knee cartilage injury model),low dose-PDS group(2 mg/kg PDS treatment)and high dose-PDS group(10 mg/kg PDS treatment).Immunohistochemistry was used to observe the histological changes;deoxynucleotidyl terminal transferase-mediated dUTP Nick-End Labeling(TUNEL)assay to detect cell apoptosis;enzyme-linked immunosorbent assay to detect matrix metalloproteinase-3(MMP-3),transforming growth factor-β1(TGF-β1)and tumor necrosis factor-α(TNF-α)levels;real-time fluorescence quantitative polymerization Enzyme chain reaction(qRT-PCR)to detect matrix metalloproteinase-13(MMP-13)and typeⅡcollagen(COL-Ⅱ)gene mRNA expression.The t test was used for comparison between two groups,and the one-way analysis of variance was used for comparison among multiple groups(LSD-t test was used for pairwise comparison between groups).Results The Mankin score,apoptosis rate,MMP-3,TGF-β1 and TNF-αlevels,and MMP-13 mRNA expression in the model group were significantly higher than those in the Sham group[Mankin score:8.93±1.05 vs.1.05±0.08;apoptosis rate:(58.28±4.24)%vs.(4.48±0.87)%;MMP-3:(33.04±3.21)vs.(8.52±1.26)pg/ml;TGF-β1:(5.09±0.68)vs.(2.27±0.31)pg/ml;TNF-α:(14.92±2.31)vs.(5.11±1.08)pg/ml;MMP-13 mRNA expression:4.68±0.51 vs.1.03±0.05],and the COL-ⅡmRNA expression was significantly lower(0.17±0.02 vs.1.07±0.03),with the differences being statistically significant(t=19.478,29.126,11.375,15.128,9.233,8.306,11.233,P<0.05).The Mankin score,apoptosis rate,MMP-3,TGF-β1 and TNF-αlevels,and MMP-13 gene mRNA expression in the L-PDS group were significantly higher than those in the H-PDS group[Mankin score:6.01±0.79 vs.3.54±0.68;apoptosis rate:(36.67±3.45)%vs.(22.89±2.26)%;MMP-3:(21.09±2.18)vs.(13.38±1.42)pg/ml;TGF-β1:(3.94±0.32)vs.(2.98±0.29)pg/ml;TNF-α:(11.35±1.68)vs.(8.33±1.02)pg/ml;MMP-13 mRNA expres
作者
王石磊
房师强
宋振
蔺伟
Wang Shilei;Fang Shiqiang;Song Zhen;Lin Wei(Department of Spine and Joint Surgery,Jinan People’s Hospital Affiliated to Shandong First Medical University,Jinan 271199,China)
出处
《中华实验外科杂志》
CAS
北大核心
2022年第3期511-514,共4页
Chinese Journal of Experimental Surgery
关键词
人参二醇型皂苷
膝关节
软骨损伤
机制
Panaxadiol saponins
Knee joint
Cartilage injury
Mechanism
