四逆汤饼灸对兔膝骨关节炎软骨中Caveolin-1/p38 MAPK信号通路蛋白表达的影响

The Effect of Sini Decoction Cake Moxibustion on the Expression of Caveolin-1/p38 MAPK Signaling Pathway Proteins in Cartilage of Rabbit Knee Osteoarthritis

目的探讨四逆汤饼灸治疗膝骨关节炎(KOA)的作用机制。方法取新西兰兔50只,随机分为对照组、模型组、p38 MAPK抑制剂(抑制剂)组、四逆汤饼灸组、四逆汤饼灸联合p38 MAPK抑制剂(联合)组,每组10只。除对照组外,其他组采用右后肢膝关节伸直位石膏管型固定制备兔KOA模型。对照组、模型组不予治疗;抑制剂组于关节腔内注射p38 MAPK抑制剂TAK-715,1次/d,治疗2周;四逆汤饼灸组取鹤顶、内外膝眼穴位予以四逆汤饼灸治疗,1次/d,治疗2周;联合组于关节腔内注射p38 MAPK抑制剂TAK-715,并在鹤顶、内外膝眼穴位予以四逆汤饼灸治疗,1次/d,治疗2周。采用HE染色观察软骨病理变化,免疫组织化学染色法和蛋白质印迹法(Western blot)检测软骨组织中Caveolin-1、p38 MAPK、MMP-13蛋白的表达。结果HE染色结果显示,模型组软骨表面明显粗糙,破坏严重,细胞排列紊乱,Mankin’s评分明显升高;抑制剂组、四逆汤饼灸组及联合组软骨表面较光滑,软骨细胞分布较均匀,Mankin’s评分降低。免疫组化与Western blot结果显示,与对照组比较,模型组软骨组织中Caveolin-1、p38 MAPK、MMP-13的表达均升高(P均<0.05);与模型组比较,抑制剂组、四逆汤饼灸组及联合组软骨组织中Caveolin-1、p38MAPK、MMP-13的表达均降低(P均<0.05)。结论四逆汤饼灸能延缓KOA软骨的进一步破坏,其机制可能与抑制Caveolin-1/p38 MAPK信号通路蛋白的表达有关。

Objective To discuss the mechanism of Sini Decoction Cake Moxibustion in the treatment of(knee osteoarthritis,KOA).Methods A total of 50 New Zealand rabbits were randomly divided into control group,model group,p38 MAPK inhibitor group(inhibitor group),Sini Decoction Cake Moxibustion group,Sini Decoction Cake Moxibustion combined with p38 MAPK inhibitor group(combined group),with 10 in each group.Except for the control group,the rabbit KOA modelswas prepared by fixing the right hind limb knee using plaster cast fixation in the other four groups.The control group and model group were not treated.The rabbits were injected with p38 MAPK inhibitor TAK-715 in the joint cavity in the inhibitor group,once a day,for 2 weeks.Heding,inner knee eyes and outer knee eyes were performed by Sini Decoction Cake Moxibustionin in the Sini Decoction Cake Moxibustion group,once a day,for 2 weeks.In the combined group,the rabbits were injected with p38 MAPK inhibitor TAK-715 in the joint cavity and Heding,inner knee eyes and outer knee eyes were performed by Sini Decoction Cake Moxibustion,once a day,for 2 weeks.After the experiment,the cartilage pathological changes were observed by Hematoxylin-eosin(HE)staining,and the expression of Caveolin-1,p38 MAPK and MMP-13 protein in cartilage tissues was detected by Immunohistochemistry and Western blot.Results HE staining results showed that the cartilage surface of the model group were obviously rough,severely damaged,the cells arrangement were disordered,and the Mankin’s score increased significantly,the cartilage surface of the inhibitor group,the moxibustion group and the combination group were smooth,the chondrocytes were evenly distributed,and the Mankin’s score was decreased.Immunohistochemistry and Western blot results showed that the expression of Caveolin-1,p38 MAPK and MMP-13 increased in the model group(P all<0.05);compared with the model group,the expression of Caveolin-1,p38 MAPK and MMP-13 decreased in the inhibitor group,moxibustion group and combination group(P all<0.05).Conclusion Sini Decoction Cake Moxibustion could delay the further destruction of KOA cartilage,and its mechanism might be related to restrain the expression levels of the Caveolin-1/p38 MAPK signaling pathway proteins.