温针灸对膝骨关节炎模型大鼠软骨组织Caspase-3、Caspase-9 mRNA转录水平的影响  

Effect of Needle-Warming Moxibustion on Transcription Level of Caspase-3 and Caspase-9 in Cartilage Tissue of Rats with Knee Osteoarthritis

作　　者：吴靖 洪昆达 WU Jing;HONG Kunda(College of Acupuncture and Moxibustion,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;The Second Affiliated Hospital of Fujian Medical University,Quanzhou 362000,China)

机构地区：[1]福建中医药大学针灸学院,福建福州350122 [2]福建医科大学附属第二医院,福建泉州362000

出　　处：《福建中医药》2024年第4期38-43,共6页Fujian Journal of Traditional Chinese Medicine

基　　金：福建省卫健委中青年骨干人才项目(2020GGA056);福建医科大学附属第二医院博士启动基金(BS202005)。

摘　　要：目的 分析温针灸对膝骨关节炎(KOA)模型大鼠软骨组织Caspase-3、Caspase-9 mRNA转录水平的影响,探讨温针灸治疗KOA的可能作用机制。方法 将24只8周龄SPF级雄性SD大鼠按照随机数字表法分为空白组、模型组与温针灸组,每组8只。模型组与温针灸组采用4%木瓜蛋白酶注射法建立KOA模型,空白组不做任何处理,当大鼠出现膝关节肿大,局部刺激有疼痛反应且Lequesne MG行为学评分≥3分时,提示造模成功。造模成功后温针灸组取双侧内膝眼、犊鼻穴,以毫针直刺,深度约5 mm,将艾段点燃放置于针柄末端,共灸2壮,留针20 min;空白组与模型组同等抓取固定20 min,1次/d,不做任何治疗。3组每周均干预6 d,休息1 d,共干预2周。干预后采用Lequesne MG行为学评分评估3组大鼠行为学表现,HE染色法及番红-固绿染色法观察3组大鼠膝关节软骨组织形态变化,Mankin’s评分评估3组大鼠软骨退变程度,TUNEL染色法观察3组大鼠膝关节软骨细胞凋亡率,qPCR检测3组大鼠膝关节软骨组织Caspase-3、Caspase-9 mRNA转录水平。结果 与空白组比较,模型组Lequesne MG行为学评分明显升高(P<0.05);HE染色结果显示膝关节软骨组织各层结构不清,表层出现缺损、裂隙,软骨细胞数量弥漫性增多或局部减少且软骨细胞排列紊乱;番红-固绿染色结果显示软骨基质番红染色不均,固绿染色范围增加,潮线不清;Mankin’s评分升高(P<0.05),膝关节软骨组织细胞凋亡率升高(P<0.05),Caspase-3mRNA转录水平无明显变化(P>0.05),Caspase-9 mRNA转录水平升高(P<0.05)。与模型组比较,温针灸组Lequesne MG行为学评分降低(P<0.05),HE染色结果显示膝关节软骨组织表层较为平整,软骨细胞数量弥漫性增多且排列不规则,但优于模型组;番红-固绿染色结果显示软骨基质番红染色未见明显异常,潮线清晰,Mankin’s评分下降(P<0.05),膝关节软骨细胞凋亡率下降(P<0.05),Caspase-3 mRNA转录水平降�Objective:To analyze the effect of needle-warming moxibustion on transcription levels of Caspase-3 and Caspase-9 in cartilage tissue of knee osteoarthritis(KOA)model rats,so that to explore the possible mechanism of needle-warming moxibustion(NWM)in the treatment of KOA.Methods:A total of 24 SPF male SD rats aged 8 weeks were randomly divided into the blank group,model group and NWM group,with 8 in each group.Establishing KOA models using 4%papain injection in the model and NWM groups,while the blank group received no treatment.The knee joint of rats enlarged and showed pain response to local stimulation,with Lequesne MG behavioral score≥3,which indicated that the KOA models were successfully established.Then the NWM group acuputured bilateral Xiyan and Dubi acupoints,the depth was about 5 mm.Placing a small section of moxa stick onto the handle of an inserted needle and igniting it.The moxibustion was carried out for twice,and the retention of needle was for 20 minutes.The blank and model groups were equally captured and fixed for 20 minutes without any treatment once a day.All the three groups were intervened for 6 days a week and rested for 1 day for 2 weeks.The Lequesne MG score was used to evaluate the behavioral performance of rats,the HE staining and the safranin-fast green staining methods were used to observe the morphological changes of knee cartilage,the Mankin's scoring system was used to evaluate the degree of cartilage degeneration,and the TUNEL staining was used to observe the apoptosis rate of cartilage cells from knee joints of rats in the three groups.The transcription level of Caspase-3 and Caspase-9 mRNA in knee cartilage were detected by qPCR.Results:Compared with the blank group,the Lequesne MG score in the model group significantly increased(P<0.05).The result of HE staining showed that the structure of all layers of the knee cartilage tissue was unclear,surface defects and cracks appeared,the number of chondrocyte diffuse increased or locally decreased,and the arrangement of chondrocytes

关 键 词：膝骨关节炎 温针灸 凋亡 CASPASE-3 CASPASE-9 

分 类 号：R245[医药卫生—针灸推拿学]