丹参注射液治疗对膝骨性关节炎模型兔关节软骨中TNF-α、IL-6、MMP-3表达的影响  

Effect of Danshen injection on expression of TNF-α,IL-6 and MMP-3 in articular cartilage of rabbits with knee osteoarthritis

作　　者：滕海[1] 陆惠英 李俊杰[2] Teng Hai;Lu Huiying;Li Junjie(Department of Orthopedics,Affiliated Central Hospital of Shenyang Medical College,Shenyang 110024,China;Department of Surgery,Affiliated Hospital of Jiaying College Medical College,Meizhou 514000,China)

机构地区：[1]沈阳医学院附属中心医院骨科,110024 [2]嘉应学院医学院附属医院外科,广东梅州514000

出　　处：《国际免疫学杂志》2020年第1期20-25,共6页International Journal of Immunology

基　　金：梅州市医药卫生科技计划面上项目(2018KY156)。

摘　　要：目的 研究丹参注射液治疗膝骨性关节炎(knee osteoarthritis,KOA)模型兔关节软骨中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素(interleukin,IL)-6、基质金属蛋白酶-3(matrix metalloproteinase-3,MMP-3)的表达情况,探讨丹参注射液治疗膝骨性关节炎的分子机制。方法 选用40只健康白兔,随机分成四组,即假手术组、KOA模型(KOA)组、KOA+透明质酸钠(sodium hyaluronate,SH)组、KOA+丹参组。假手术组只作切口,不切断半月板,其余组均采用切断兔右膝关节半月板的方式,建立一个兔关节失稳的实验动物模型。KOA组给予0.9%生理盐水(0.7 mL/d),KOA+SH组给予兔关节内注射SH(0.4 mL/d),KOA+丹参组给予丹参注射液(0.7 mL/d),均连续注射5周。采用矿场实验验证兔失稳模型建立是否对兔的运动能力产生影响,并采用聚合酶链反应(polymerase chain reaction,PCR)和Western blot法分别检测兔膝关节软骨中TNF-α、IL-6、MMP-3的mRNA含量和蛋白表达量。酶联免疫吸附剂测定(enzyme linked immunosorbent assay,ELISA)检测兔膝关节液中TNF-α、IL-6、MMP-3的含量。结果 在KOA模型建立之前,各实验组动物均无明显的运动能力差异,模型建立之后,KOA组兔子运动的距离明显低于假手术组[(150±5)比(580±9),t=60.610,P<0.05],而KOA+丹参组与KOA组相比,家兔移动的距离明显增加,差异具有统计学意义[(438±27)比(150±5),t=16.730,P<0.05]。KOA组兔关节液中TNF-α、IL-6和MMP-3含量明显高于假手术组[(22.62±2.18)mg/mL比(11.74±2.09)mg/mL,(4.01±0.14)mg/mL比(1.76±0.11)mg/mL,(0.57±0.05)mg/mL比(0.27±0.03)mg/mL,t值分别为4.413、17.620、6.495,P值均<0.05];KOA+丹参组兔关节液中TNF-α、IL-6、MMP-3含量较KOA组明显降低,差异具有统计学意义[(15.01±2.37)mg/mL比(22.62±2.18)mg/mL,(2.47±0.19)mg/mL比(4.01±0.14)mg/mL,(0.40±0.02)mg/mL比(0.57±0.05)mg/mL,t值分别为3.725、8.803、9.185,P值均<0.05]。在兔膝关节软骨的mRNA的检测中,KOA组TNFObjective To study the expression of tumor necrosis factor-α(TNF-α),interleukin(IL)-6,matrix metalloproteinase-3(MMP-3)in the articular cartilage of rabbits with knee osteoarthritis(KOA)treated by Danshen injection,and to explore the molecular mechanism of Danshen injection in the treatment of knee osteoarthritis.Methods Forty healthy white rabbits were randomly divided into four groups:sham operation group,KOA model(KOA)group,KOA+sodium hyaluronate(SH)group and KOA+Danshen group.Sham operation group only made incision without cutting meniscus.The other groups all used cutting meniscus of rabbit right knee joint to establish an experimental animal model of joint instability.The KOA group was given 0.9%normal saline(0.7 mL/d),the KOA+SH treatment group was given intra-articular injection of SH(0.4 mL/d),and the KOA+Danshen treatment group was given Danshen injection(0.7 mL/d)for 5 weeks.The field experiment was used to verify whether the instability model of rabbits could affect the motor ability of rabbits.The mRNA content and the protein expression of TNF-α,IL-6 and MMP-3 were detected separately by polymerase chain reaction(PCR)and Western blot.The contents of TNF-α,IL-6 and MMP-3 in knee joint fluid were detected by enzyme linked immunosorbent assay(ELISA)kit.Results Before the establishment of KOA model,there was no significant difference in motor ability among the groups.After the establishment of the model,the movement distance of rabbits in the KOA group was significantly lower than that in the sham operation group[(150±5)vs(580±9),t=60.610,P<0.05].The walking distance in the KOA+Danshen group was significantly increased than that in the KOA group[(438±27)vs(150±5),t=16.730,P<0.05].In rabbit joint fluid,the contents of TNF-α,IL-6,MMP-3 in KOA group were significantly higher than those in sham operation group[(22.62±2.18)mg/mL vs(11.74±2.09)mg/mL,(4.01±0.14)mg/mL vs(1.76±0.11)mg/mL,(0.57±0.05)mg/mL vs(0.27±0.03)mg/mL,t values were 4.413,17.620 and 6.495 respectively,all P values<0.05],while,c

关 键 词：膝骨性关节炎 丹参注射液 肿瘤坏死因子-α 白细胞介素-6、基质金属蛋白酶-3 

分 类 号：R-332[医药卫生] R28