中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (12): 2475-2483.doi: 10.12307/2025.394

• 软骨组织构建 cartilage tissue construction • 上一篇    下一篇

艾灸调节circPan3/miR-667-5p/ghrelin信号通路减轻膝骨关节炎大鼠的软骨病变

廖  晴1,曾  静2,陈  峻3,袁立霞1,刘  刚3   

  1. 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



  • 收稿日期:2024-05-13 接受日期:2024-06-21 出版日期:2025-04-28 发布日期:2024-09-09
  • 通讯作者: 刘刚,博士,副教授,南方医科大学南方医院康复科,广东省广州市 510000
  • 作者简介:廖晴,女,1992年生,湖南省常宁市人,汉族,南方医科大学在读博士,主要从事膝骨关节炎作用机制的研究。
  • 基金资助:
    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

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

Liao Qing1, Zeng Jing2, Chen Jun3, Yuan Lixia1, Liu Gang3   

  1. 1School of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510000, Guangdong Province, China; 2Southern University of Science and Technology Hospital, Shenzhen 518000, Guangdong Province, China; 3Department of Rehabilitation, Nanfang Hospital, Southern Medical University, Guangzhou 510000, Guangdong Province, China
  • Received:2024-05-13 Accepted:2024-06-21 Online:2025-04-28 Published:2024-09-09
  • Contact: Liu Gang, MD, Associate professor, Department of Rehabilitation, Nanfang Hospital, Southern Medical University, Guangzhou 510000, Guangdong Province, China
  • About author:Liao Qing, MD candidate, School of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510000, Guangdong Province, China
  • Supported by:
    the National Natural Science Foundation of China, Nos. 82247619 (to LG) and 82274400 (to YLX)

摘要:


文题释义:
Ghrelin:是一种由28个氨基酸组成的肽类激素,可增强软骨自噬、抑制炎症和软骨退变,预防膝骨关节炎的发生和发展。
自噬:是一种高度保守的代谢降解过程,细胞通过自噬降解和回收利用细胞内生物大分子和受损细胞器以维持体内平衡,自噬在维持软骨细胞稳态、保护软骨中有着至关重要的作用。

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 < 0.000 1);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 < 0.000 1)。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表达减少(P < 0.01);与模型组相比,艾灸可逆转上述指标改变(P < 0.01)。③结果说明艾灸可能通过调节circPan3/miR-667-5p/Ghrelin信号通路增强软骨细胞自噬,抑制膝骨关节炎炎症和软骨破坏,对软骨发挥保护作用。
https://orcid.org/0000-0002-7227-2676(廖晴)   


中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程

关键词: 膝骨关节炎, 艾灸, ghrelin, 软骨, 自噬, 大鼠, 信号通路

Abstract: 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 cells, narrowing of the joint space, rough and uneven cartilage surfaces, thinning, reduction in the number of chondrocytes, a significant increase in synovitis and Osteoarthritis Research Society International scores, and a significant up-regulation of interleukin-6 and tumor necrosis factor-α levels (P < 0.000 1). In comparison, the sham operation group showed no significant changes. Compared with the model group, the moxibustion group exhibited a reduction in synovial thickening, a decrease in inflammatory cell infiltration, an increase in joint space, a reduction in cartilage roughness and loss of integrity, an increase in the number of chondrocytes, a reduction in synovitis and Osteoarthritis Research Society International scores, and a significant reduction in the levels of interleukin-6 and tumor necrosis factor-α (P < 0.000 1). The results of in vitro experiments demonstrated that the expression of matrix metalloproteinase 13, ADAMTS5, and miR-667-5P was elevated, while the expression of type II collagen, Sox9, autophagy markers, Ghrelin, and Circpan3 was reduced in the model group compared with the normal control group (P < 0.01). Moxibustion was found to reverse these changes in the moxibustion group compared with the model group (P < 0.01). To conclude, moxibustion may enhance chondrocyte autophagy by modulating the circPan3/miR-667-5p/Ghrelin signaling pathway, which may inhibit knee osteoarthritis inflammation and cartilage destruction, exerting a protective effect on cartilage.

中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程

Key words: knee osteoarthritis, moxibustion, ghrelin, cartilage, autophagy, rat, signaling pathway

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