来源：风湿病与关节炎,2023,12（10）:6-10.

火针对膝骨关节炎大鼠关节软骨细胞自噬相关蛋白LC3及血清炎症因子的影响（基础研究）

高仰来，冯　鹏，李晓霞

　　【摘　要】目的：观察火针对膝骨关节炎大鼠膝关节软骨细胞自噬相关蛋白微管相关蛋白1轻链3（LC3）及血清炎症因子的影响。方法：将36只健康SD大鼠随机分为空白对照组（8只）和造模组（28只），采用改良Hulth法制备膝骨关节炎模型，6周后随机选取造模组4只大鼠处死，行关节软骨病理学检查判定模型制备是否成功，随后将剩余24只大鼠随机分为模型对照组、西药组、火针组，每组8只。火针组给予火针治疗干预，每周2次；西药组给予盐酸氨基葡萄糖片溶液1.6 mL（0.384 mg）灌胃，每日1次；空白对照组、模型对照组给予等量生理盐水灌胃，每日1次。各组均以4周为1个疗程。1个疗程结束后，采集各组大鼠血清、关节软骨，采用ELISA法、PCR法对血清标本的基质金属蛋白酶（MMP）-1、MMP-3、MMP-13、肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）、LC3水平进行检测；HE染色法观察关节软骨组织形态结构。结果：与空白对照组比较，模型对照组、西药组、火针组MMP-1、MMP-3、MMP-13、TNF-α、IL-1β表达升高（P < 0.05），LC3表达降低（P < 0.05）；与模型对照组比较，火针组MMP-1、MMP-3、MMP-13、TNF-α、IL-1β表达降低（P < 0.05），LC3表达升高（P < 0.05）；与西药组比较，火针组MMP-1、MMP-3、MMP-13、TNF-α、IL-1β表达降低（P < 0.05），LC3表达升高（P < 0.05）。关节软骨病理切片显示，火针组关节腔隙明显，软骨表面毛糙，滑膜组织有少量破损，炎性细胞浸润但少于模型对照组，部分潮线破坏，软骨细胞排列尚均匀。结论：火针可降低血清炎性因子含量，上调自噬标志性蛋白LC3的表达，抑制膝骨关节炎炎症反应，缓解炎性浸润，减轻软骨细胞的破坏，改善其病理状态。

　　【关键词】　膝骨关节炎；火针；自噬；微管相关蛋白1轻链3；炎症因子；大鼠

The Effect of Fire Acupuncture on Autophagy Related Protein LC3 and Serum Inflammatory Factors in Joint Chondrocytes of Rats with Knee Osteoarthritis

GAO Yang-lai,FENG Peng,LI Xiao-xia

　　【ABSTRACT】Objective:To observe the effect of fire acupuncture on autophagy related protein LC3 and serum inflammatory factors in knee joint chondrocytes of rats with knee osteoarthritis.Methods:Thirty-six healthy SD rats were randomly divided into a blank control group（8 rats）and a modeling group（28 rats）.Knee osteoarthritis models were prepared using the improved Hulth method.After 6 weeks,4 rats in the modeling group were randomly selected and killed.Joint cartilage pathology examination was performed to determine whether the model was successful.Subsequently,the remaining 24 rats were randomly divided into a model control group,a Western medicine group,and a fire acupuncture group,with eight rats in each group.The fire acupuncture group received fire acupuncture treatment intervention,twice a week;the western medicine group was given 1.6mL（0.384 mg）of glucosamine hydrochloride tablet solution by gavage,once a day;the blank control group and the model control group were given an equal amount of physiological saline by gavage,once a day.A treatment course of four weeks was for each group.After one course of treatment,collect serum and articular cartilage from rats in each group,and use ELISA and PCR methods to detect the levels of MMP-1,MMP-3,MMP-13,TNF-α,IL-1β and LC3;HE staining method was used to observe the morphology and structure of articular cartilage tissue.Results:Compared with the blank control group,the expressions of MMP-1,MMP-3,MMP-13,TNF-α and IL-1β in the model control group,the western medicine group and the fire acupuncture group increased（P < 0.05）,while the expression of LC3 decreased（P < 0.05）;compared with the model control group,the expressions of MMP-1,MMP-3,MMP-13,TNF-α and IL-1β in the fire acupuncture group decreased（P < 0.05）,while the expression of LC3 increased（P < 0.05）;compared with the western medicine group,the expressions of MMP-1,MMP-3,MMP-13,TNF-α and IL-1β in the fire acupuncture group decreased（P < 0.05）,while the expression of LC3 increased（P < 0.05）.Pathological sections of articular cartilage showed that in the fire acupuncture group,the joint cavity was obvious,the surface of the cartilage was rough,there was a small amount of damage to synovial tissue,inflammatory cell infiltration was less than that in the model control group,some tidal lines were damaged,and the arrangement of chondrocytes was still uniform.Conclusion:Fire acupuncture can reduce the content of serum inflammatory factors,upregulate the expression of autophagy marker protein LC3,inhibit the inflammatory response of knee osteoarthritis,alleviate inflammatory infiltration and chondrocyte damage,and improve the pathological state.

　　【Keywords】 knee osteoarthritis;fire acupuncture;autophagy;LC3;inflammatory factors;rats