艾灸调节circ Pan3/mi R-667-5p/Ghrelin信号通路减轻膝骨关节炎大鼠的软骨病变  

Moxibustion alleviates cartilage lesions in rats with knee osteoarthritis through regulating the circPan3/miR-667-5p/Ghrelin signaling pathway

作　　者：廖晴 曾静 陈峻 袁立霞[1] 刘刚[3] Liao Qing;Zeng Jing;Chen Jun;Yuan Lixia;Liu Gang(School of Traditional Chinese Medicine,Southern Medical University,Guangzhou 510000,Guangdong Province,China;Southern University of Science and Technology Hospital,Shenzhen 518000,Guangdong Province,China;Department of Rehabilitation,Nanfang Hospital,Southern Medical University,Guangzhou 510000,Guangdong Province,China)

机构地区：[1]南方医科大学中医药学院,广东省广州市510000 [2]南方科技大学医院,广东省深圳市518000 [3]南方医科大学南方医院康复科,广东省广州市510000

出　　处：《中国组织工程研究》2025年第12期2475-2483,共9页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(82247619),项目负责人:刘刚;国家自然科学基金项目(82274400),项目负责人:袁立霞。

摘　　要：背景:既往研究表明艾灸能减轻膝骨关节炎软骨病变,延缓膝骨关节炎进展;课题组前期研究中发现调控circPan3/miR-667-5p/Ghrelin信号通路可增强软骨细胞自噬水平,从而在膝骨关节炎中发挥软骨保护作用。目的:基于circPan3/miR-667-5p/Ghrelin信号通路探究艾灸对大鼠膝骨关节炎软骨的影响。方法:SD大鼠随机分为假手术组、模型组和艾灸组(n=10),对后2组大鼠行右后肢膝关节前交叉韧带横断和内侧半月板部分切除术建立膝骨关节炎大鼠模型;假手术组大鼠仅接受关节囊切开术。6周后,艾灸组大鼠每日艾灸“足三里”“肾俞”穴区20 min,持续4周,其余2组不予以特殊处理。4周后通过组织病理学染色观察膝关节滑膜病变和软骨损伤程度,通过酶联免疫吸附实验检测血清中炎症因子白细胞介素6和肿瘤坏死因子α的变化。此外,取上述3组大鼠右侧膝关节软骨细胞分为:正常对照组、模型组和艾灸组,通过Western blot和qRT-PCR检测各组细胞中软骨基质合成代谢分子(Ⅱ型胶原蛋白和Sox9)、基质降解分子(基质金属蛋白酶13和ADAMTS5)、自噬标志物(ATG5、Beclin-1、微管相关蛋白1轻链3)、Ghrelin、CircPan3以及miR-667-5P的表达水平。结果与结论:①治疗4周后,相对于假手术组,模型组大鼠滑膜显著增厚,炎症细胞浸润显著,关节间隙狭窄,软骨表面粗糙不平,变薄,软骨细胞数量减少,滑膜炎和OARSI评分显著升高,白细胞介素6和肿瘤坏死因子α水平明显上调(P<0.0001);与模型组相比,艾灸组滑膜增厚减轻,炎症细胞浸润减少,关节间隙增宽,软骨更光滑完整、变厚,软骨细胞数量增多,滑膜炎和OARSI评分降低,白细胞介素6和肿瘤坏死因子α水平显著降低(P<0.0001)。②体外实验结果表明,与正常对照组相比,模型组软骨细胞基质金属蛋白酶13、ADAMTS5和miR-667-5P表达增多,Ⅱ型胶原蛋白、Sox9、自噬标志物、Ghrelin和Circpan3表达减�BACKGROUND:Previous studies have demonstrated that moxibustion can alleviate osteoarthritis-induced cartilage lesions and delay the progression of knee osteoarthritis.In a previous study conducted by our research group,it was found that regulating the circPan3/miR-667-5p/Ghrelin signaling pathway can enhance the level of autophagy in chondrocytes,thereby exerting a cartilage protective role in knee osteoarthritis.OBJECTIVE:To explore the effect of moxibustion on knee osteoarthritis cartilage in rats based on the circPan3/miR-667-5p/Ghrelin signaling pathway.METHODS:Sprague-Dawley rats were randomly divided into three groups(n=10 per group):sham operation,model,and moxibustion groups.A rat model of knee osteoarthritis was established in the latter two groups by anterior cruciate ligament transection and medial meniscus resection.Rats in the sham operation group underwent arthrotomy only.At 6 weeks after modeling,moxibustion intervention at Zusanli and Shenyu acupoints was applied in the moxibustion group,20 minutes daily for 4 weeks,while the other two groups did not receive any special treatment.After 4 weeks,histopathological staining was performed to observe the degree of synovial lesions and cartilage damage in the knee joint.Additionally,changes in the serum levels of inflammatory cytokines,interleukin-6 and tumor necrosis factor-α,were detected by enzyme-linked immunosorbent assay.Furthermore,chondrocytes isolated from the above three groups of rats were divided into three groups:a normal control group,a model group,and a moxibustion group.The expression levels of cartilage matrix anabolic molecules(type II collagen and Sox9),matrix degradation molecules(matrix metalloproteinase 13 and ADAMTS5),autophagy markers(ATG5,Beclin-1,microtubule-associated protein 1 light chain 3),Ghrelin,CircPan3,and miR-667-5P were determined by western blot and qRT-PCR in each group.RESULTS AND CONCLUSION:After 4 weeks of treatment,the model group exhibited significant thickening of the synovium,infiltration of inflammatory cell

关 键 词：膝骨关节炎 艾灸 GHRELIN 软骨 自噬 大鼠 信号通路 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R496