温针灸对兔膝骨关节炎软骨中ASC、Caspase-1和P2X7蛋白表达的影响  

The Effect of Warm Acupuncture and Moxibustion on the Expressions of ASC,Caspase-1 and P2X7 in Cartilage of Rabbit Knee Osteoarthritis

作　　者：王亭 刘娣[2] 刘君伟 李龙[2] 武永利[2,3] WANG Ting;LIU Di;LIU Junwei;LI Long;WU Yongli(School of Traditional Chinese Medicine,Ningxia Medical University,Yinchuan 750004,China;Department of Orthopedics and Traumatology of Traditional Chinese Medicine,General Hospital of Ningxia Medical University,First Clinical Medical College of Ningxia Medical University,Yinchuan 750004,China;Ningxia Hui Autonomous Region Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of High Incidence,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学中医学院,银川750004 [2]宁夏医科大学总医院中医骨伤科,宁夏医科大学第一临床医学院,银川750004 [3]宁夏回族自治区区域高发病中西医结合防治研究重点实验室,银川750004

出　　处：《宁夏医科大学学报》2024年第4期406-412,共7页Journal of Ningxia Medical University

基　　金：国家自然科学基金项目(82160939);宁夏回族自治区第二批科技领军人才项目(KJT201610)。

摘　　要：目的 观察温针灸对膝骨关节炎(KOA)模型兔关节软骨组织中凋亡相关斑点样蛋白(ASC)、半胱氨酸蛋白酶-1(Caspase-1)、嘌呤能离子通道型受体7(P2X7)表达的影响,研究温针灸治疗KOA的作用机制。方法 取40只雄性新西兰兔随机分为空白组、模型组、抑制剂组和温针灸组,每组10只。除空白组外,其余各组采用经典右后肢膝关节伸直位,用石膏管型固定4周制备兔KOA模型。造模成功后,各组给予相应干预措施。应用Lequesne MG评分量表评估兔膝关节状态改变与行为学改变,HE染色观察膝关节软骨病理形态学改变,免疫组织化学法和Western blot法检测软骨组织中ASC、Caspase-1、P2X7蛋白的表达,免疫荧光检测Caspase-1蛋白的表达。结果 造模前,各组实验兔Lequesne MG评分差异无统计学意义(P>0.05);造模后,与空白组相比,其余组Lequesne MG评分均上升(P<0.05);干预治疗后,与模型组比较,温针灸组和抑制剂组评分降低(P<0.05)。HE染色显示,空白组软骨表面结构完整,软骨细胞排列均匀;模型组软骨表面粗糙不平,软骨受损明显,软骨细胞形态大小不一,排列紊乱;抑制剂组和温针灸组软骨表面相对光滑、完整,软骨细胞分布较为均匀,软骨层结构相对清晰。与空白组比较,模型组Mankin’s评分升高(P<0.05);与模型组比较,温针灸组和抑制剂组Mankin’s评分降低(P<0.05)。免疫组织化学法、免疫荧光及Western blot检测结果显示,与空白组比较,模型组ASC、Caspase-1、P2X7的表达均升高(P<0.05);与模型组比较,温针灸组和抑制剂组ASC、Caspase-1、P2X7的表达均降低(P<0.05);温针灸组和抑制剂组相比,ASC、Caspase-1、P2X7表达差异均无统计学意义(P均>0.05)。结论 温针灸可减轻兔KOA软骨损伤,其机制可能与ASC、Caspase-1、P2X7表达降低及细胞焦亡抑制有关。Objective To investigate the impact of warm acupuncture and moxibustion on the expressions of ASC,Caspase-1 and P2X7 in cartilage tissue of rabbit model of knee osteoarthritis(KOA),and to study the mechanism of warm acupuncture and moxibustion on KOA.Methods A total of forty male New Zealand rabbits were randomly assigned into blank group,model group,inhibitor group and warm acupuncture and moxibustion group,with 10 rabbits in each group.Except blank group,the remaining groups were prepared by the classical plaster tube fixation in the right posterior extension of the knee joint for 4 weeks to establish the rabbit KOA models.After successful modeling,each group was given the corresponding intervention measures.Lequesne MG rating scale was used to evaluate the change of rabbits knee joint state and behavior.HE staining was taken to observe the pathological morphology of rabbit knee cartilage.Immunohistochemistry and Western blot were used to detect the expression levels of ASC,Caspase-1 and P2X7 protein in cartilage tissue.Immunofluorescence was used to detect the expression of Caspase-1 protein.Results Lequesne MG scores showed that there was no significant difference among all groups before modeling(P>0.05).After modeling,Lequesne MG scores of other groups increased significantly compared with the blank group(P<0.05).Following the intervention,the scores decreased significantly in the warm acupuncture and moxibustion group and the inhibitor group compared with the model group(P<0.05).The HE staining results showed that the surface structure of the cartilage in the blank group was complete,and the cartilage cells were uniform.The cartilage surface of the model group was rough and uneven,the cartilage was damaged obviously,the shape of the cartilage cells were different,and the arrangement of the cartilage cells was disordered.In the warm acupuncture and moxibustion group and the inhibitor group,the surface of cartilage is relatively smooth and complete,and the distribution of cartilage cells was relatively unifor

关 键 词：温针灸 膝骨关节炎 凋亡相关斑点样蛋白 半胱氨酸蛋白酶-1 嘌呤能离子通道型受体7 

分 类 号：R246.9[医药卫生—针灸推拿学]