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Effects of different moxibustion time on knee cartilage morphology and the expression of TNF-α and IL-10 in rats with knee osteoarthritis

不同施灸时间对膝骨关节炎大鼠膝关节软骨组织形态及TNF-α和IL-10表达的影响

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Abstract

Objective

To observe the effects of different moxibustion time on cartilage morphology, tumor necrosis factor (TNF)-α and interleukin (IL)-10 of the knee joint in rats with knee osteoarthritis (KOA), and to explore the best treatment time of moxibustion for KOA.

Methods

Healthy male Wistar rats were randomly divided into a blank group, a model group, a 15-minute-moxibustion group, a 30-minute-moxibustion group, and a 60-minute-moxibustion group, with 10 rats in each group. Except for the blank group, the KOA model was established in all groups by injecting sodium iodoacetate solution into the knee joint cavity of rats. Rats in the 15-minute-moxibustion group, the 30-minute-moxibustion group, and the 60-minute-moxibustion group were all treated with mild moxibustion intervention for 15 min, 30 min, and 60 min, respectively at Neixiyan (EX-LE4) and Dubi (ST35) points near the patella, 3 times a week for 4 weeks, 12 times in total. Rats in the blank group and the model group were fixed for 30 min without moxibustion intervention. Macroscopic observation for the smoothness of knee cartilage surface was performed after the intervention. Hematoxylin-eosin staining, toluidine blue staining, and Mankin score were used to evaluate the pathological changes in the cartilage. The expression levels of TNF-α and IL-10 in the serum were detected by enzyme-linked immunosorbent assay.

Results

Compared with the blank group, the articular cartilage surface in the model group was rough, the chondrocyte arrangement was irregular, the Mankin score and the serum TNF-α expression were significantly increased (P<0.05), while the expression of serum IL-10 was significantly decreased (P<0.05). Compared with the model group, the articular cartilage surface was smoother, the chondrocytes were arranged neatly, the Mankin score and serum TNF-α expression level were significantly lower in the three moxibustion intervention groups (P<0.05); the serum IL-10 level in the 30-minute-moxibustion group and the 60-minute-moxibustion group was increased significantly (P<0.05). Compared with the 15-minute-moxibustion group, the articular cartilage surface in the 30-minute-moxibustion group and the 60-minute-moxibustion group was smoother, the chondrocyte arrangement was more regular, the Mankin score and the serum TNF-α level were decreased significantly (P<0.05), and the serum IL-10 level was increased (P<0.05). There was no significant difference in the serum TNF-α or IL-10 level between the 30-minute-moxibustion group and the 60-minute-moxibustion group (P>0.05).

Conclusion

Moxibustion can obviously improve the morphology and structure of KOA articular cartilage, protect articular cartilage, inhibit cartilage inflammation, and delay KOA cartilage degeneration. Moxibustion’s effect is closely related to moxibustion time; the therapeutic effect of the 30-minute-moxibustion and the 60-minute- moxibustion is better than that of the 15-minute-moxibustion.

摘要

目的

观察艾灸不同施灸时间对膝骨关节炎(KOA)大鼠膝关节软骨形态学、肿瘤坏死因子(TNF)-α和白细胞介素(IL)-10 的影响, 探究艾灸治疗KOA的最佳治疗时长。

方法

将健康雄性Wistar大鼠根据随机数字表法分为空白组、模型组、艾灸15 min组、艾灸30 min组和艾灸60 min组, 每组10 只。除空白组外, 其余组采用向大鼠膝关节腔内注射碘乙酸钠溶液的方法复制KOA模型。艾灸15 min组、艾灸30 min组和艾灸60 min组均采用温和灸干预, 干预位置均为髌骨附**的内膝眼穴及犊鼻穴, 干预时间分别为15 min、30 min和60 min, 每周干预3 次, 连续4 周, 共12 次。空白组和模型组大鼠在其他三组艾灸干预的同时固定30 min, 不予艾灸干预。干预结束后, 肉眼观察膝关节软骨表面光滑度; 使用苏木素-伊红染色、甲苯胺蓝染色和Mankin评分评价软骨病理改变程度; 通过酶联免疫吸附法检测血清中TNF-α和IL-10 表达变化。

结果

与空白组比较, 模型组大鼠关节软骨表面粗糙, 软骨细胞排列欠规整, Mankin评分及血清TNF-α的表达明显增高(P<0.05), 血清IL-10 的表达明显降低(P<0.05)。与模型组比较, 3 个艾灸干预组大鼠关节软骨表面较光滑, 软骨细胞排列较整齐, Mankin评分及血清TNF-α的表达明显降低(P<0.05); 艾灸30 min组和艾灸60 min组血清IL-10 的表达明显增高(P<0.05)。与艾灸15 min组比较, 艾灸30 min组和艾灸60 min组大鼠关节软骨表面较光滑, 软骨细胞排列较整齐, Mankin评分及血清TNF-α的表达明显降低(P<0.05), 血清IL-10 的表达增高(P<0.05)。艾灸30 min组与艾灸60 min组比较, 血清TNF-α及IL-10 的表达差异无统计学意义(P>0.05)。

结论

艾灸能明显改善KOA关节软骨的形态结构, 保护关节软骨, 抑制软骨炎症, 延缓KOA软骨退变。其作用效果与艾灸治疗时间密切相关, 艾灸30 min与艾灸60 min的治疗效果优于艾灸15 min。

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Acknowledgments

This work was supported by the Scientific Research Project of Hebei Provincial Administration of Traditional Chinese Medicine (河北省中医药管理局科研计划项目, No. 2020135).

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Authors and Affiliations

  1. Hebei University of Chinese Medicine, Shijiazhuang, 050091, China

    Qing Li  (**情), **ju Xue  (薛**聚), **aoqin Zhang  (张小琴), Yejuan Jia  (贾叶娟), **g Xu  (徐晶), Zilong Chen  (陈子龙), Chunsheng Jia  (贾春生) & Haijiao **ng  (邢海娇)

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  1. Qing Li  (**情)
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  2. **ju Xue  (薛**聚)
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  4. Yejuan Jia  (贾叶娟)
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Correspondence to **ju Xue  (薛**聚) or Haijiao **ng  (邢海娇).

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Li, Q., Xue, P., Zhang, X. et al. Effects of different moxibustion time on knee cartilage morphology and the expression of TNF-α and IL-10 in rats with knee osteoarthritis. J. Acupunct. Tuina. Sci. 21, 187–196 (2023). https://doi.org/10.1007/s11726-023-1375-3

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