摘要
目的:探究富血小板血浆(PRP)和弗林蛋白酶(Furin)激动剂对SD大鼠膝骨关节炎(KOA)的影响。方法:①体内实验部分,通过手术离断SD大鼠前交叉韧带建立KOA模型;实验组(10只)关节腔隔天注射0.5ml激活后的PRP,对照组(模型组,10只)和假手术组(10只)关节腔隔天注射0.9%氯化钠溶液0.5ml,持续4周;取大鼠膝关节组织采用HE、甲苯胺蓝和番红固绿染色显示病理损伤,采用免疫组化染色检测软骨蛋白聚糖抗体(ADAMTS)-5和白细胞介素(IL)-1β两种相关抗体的表达。②体外实验部分,采用脂多糖(LPS)诱导关节软骨细胞炎症模型,用递增浓度PRP和Furin激动剂干预LPS处理的SD大鼠软骨细胞,采用实时荧光定量PCR(RT-PCR)检测相关基因[基质金属蛋白酶(MMP)-3、MMP-13、IL-6、ADAMTS-4、ADAMTS-5、IL-1β和环氧化酶(COX)-2、聚蛋白多糖(ACAN)和Ⅱ型胶原A1(COL2A1)]mRNA的表达,采用酶联免疫吸附(ELISA)法检测IL-1β、IL-6、肿瘤坏死因子-α(TNF-α)和诱导型一氧化氮合酶(iNOS)水平。结果:①体内实验部分,膝关节大体观及病理切片染色结果均显示,与对照组相比,实验组关节软骨病理损伤程度明显减轻。②体外实验部分,ELISA结果显示,加入LPS后IL-1β、IL-6、TNF-α、iNOS浓度显著增加,加入不同浓度PRP后各炎症因子浓度随着PRP浓度的增加呈下降趋势,加入Furin激动剂后各炎症因子浓度亦有所下降(均P<0.05);RT-PCR结果显示,加入LPS后MMP-3、MMP-13、IL-6、ADAMTS-4、ADAMTS-5、IL-1β和COX-2的mRNA表达增加,ACAN和COL2A1的mRNA表达下降(均P<0.05),而加入不同浓度PRP和Furin激动剂后上述指标有所修复。结论:PRP和Furin激动剂对SD大鼠KOA均有明显的缓解作用。
Objective:To investigate the remission effect of knee osteoarthritis(KOA)in SD rats by platelet-rich plasma(PRP)and Furin agonists.Methods:①In vivo experiment,the KOA model was established by surgical disconnection of the anterior cruciate ligament of SD rats;the experimental group(10 rats)was injected 0.5 ml of activated PRP into the joint cavity every other day,and the control group(10 rats)and the sham group(10 rats)were injected 0.5 ml of 0.9%sodium chloride solution every other day for 4 weeks;the knee joint tissues of rats were stained with HE,toluidine blue and fan-red solid green to show pathological damage;immunohistochemical staining was used to detect the expression of ADAMTS-5 and IL-1βantibodies.②In vitro experiment,the articular chondrocyte inflammation model was induced by LPS,and increasing concentrations of PRP and Furin agonist were used to interfere with LPS-treated SD rat chondrocytes;the mRNA expressions of related genes(MMP-3,MMP-13,IL-6,ADAMTS-4,ADAMTS-5,IL-1β,COX-2,ACAN and COL2A1)were detected by RT-PCR;the levels of IL-1β,IL-6,TNF-αand iNOS were measured by ELISA.Results:①In vivo experiment,the gross view of knee joint and the staining results of pathological sections showed that compared with control group,the pathological injury degree of articular cartilage in experimental group was significantly reduced.②In vitro experiment,ELISA results showed that the concentrations of IL-1β,IL-6,TNF-αand iNOS increased significantly after adding LPS,and the concentrations of inflammatory factors decreased with the increase of PRP concentration after adding different concentrations of PRP,and the concentrations of inflammatory factors also decreased after adding Furin agonist(all P<0.05);RT-PCR results showed that the mRNA expressions of MMP-3,MMP-13,IL-6,ADAMTS-4,ADAMTS-5,IL-1βand COX-2 increased after adding LPS,and the mRNA expressions of ACAN and COL2A1 decreased(all P<0.05),while the above indexes were repaired after adding different concentrations of PRP and Furin agonist.Conclusion:Both PRP and Furin agonist have significant alleviating effect on KOA in SD rats.
作者
周安远
梁朝鑫
杨斯淇
吕沛
杨帆
杨彪
吴立蔚
王哲纬
杨渊
ZHOU Anyuan;LIANG Chaoxin;YANG Siqi;LYU Pei;YANG Fan;YANG Biao;WU Liwei;WANG Zhewei;YANG Yuan(Guangxi Medical University,Nanning 530021,China)
出处
《陕西医学杂志》
CAS
2022年第9期1043-1048,1054,共7页
Shaanxi Medical Journal
基金
国家重点研发计划项目(2020YFB1313801)
广西壮族自治区自然科学基金资助项目(2019JJA140664)
南宁市青秀区科技计划项目(2020014)。
