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目的:观察短刺加电针对膝骨关节炎模型兔软骨Ⅱ型胶原(collagen typeⅡ)—盘状结构域受体(discoidin domain receptor,DDR)-2—基质金属蛋白酶(matrix metalloproteinase,MMP)-13信号通路的影响,探讨该治疗方法修复膝关节软骨细胞外基质的可能作用机制。方法:将40只新西兰大白兔按比例随机分为正常组(10只)和造模组(30只),造模组动物采用HulthTelhag法手术复制膝骨关节炎模型,X线评估造模结果。将造模成功的动物随机分为3组,即模型组、短刺组、普通针刺组,每组10只。短刺组采用短刺加电针法,普通针刺组采用常规针刺加电针法治疗,均选取“内膝眼”“犊鼻”“阴陵泉”“足三里”和“梁丘”进行针刺。正常组和模型组正常抓取、固定,不予以干预。每次20min,每日1次,5d为一疗程,每个疗程间间隔2d,共治疗4个疗程。疗程结束后,采用蛋白免疫印迹法(Western Blot,WB)检测软骨DDR2、Ⅱ型胶原的蛋白表达;免疫组化法检测软骨DDR2、Ⅱ型胶原、MMP13的活性;逆转录聚合酶链式反应(reverse transcription-polymerase chain reaction,RT-PCR)检测DDR2和MMP13的mRNA表达。结果:与正常组相比,其他各组动物的Ⅱ型胶原蛋白及活性表达均明显减少(均P<0.01),DDR2、MMP13活性及mRNA表达均明显增多(均P<0.01);与模型组相比,短刺组和普通针刺组中Ⅱ型胶原蛋白及活性表达显著增多(均P<0.01),DDR2、MMP13活性及mRNA表达显著减少(均P<0.01);与普通针刺组相比,短刺组中MMP13、DDR2活性及mRNA表达下降(均P<0.01),Ⅱ型胶原蛋白及活性表达明显上升(P<0.01)。结论:短刺加电针法与普通电针法均可促进膝关节软骨的修复,且短刺加电针法具有一定的优势,其作用机制可能与阻断Ⅱ型胶原/DDR2/MMP13通路的恶性循环、抑制软骨细胞外基质的持续降解有关。
OBJECTIVE: To observe the effects of pricking plus electroporation on the expression of collagen typeⅡ-discoidin domain receptor (DDR) -2-MMP (matrix metalloproteinase) -13 Signaling pathways to explore the treatment of knee cartilage repair of extracellular matrix may be the mechanism of action. Methods: Forty New Zealand white rabbits were randomly divided into normal group (n = 10) and model group (n = 30). The model group was given knee osteoarthritis by Hulth Telhag method and the model was evaluated by X-ray. The successful animals were randomly divided into three groups, model group, short stab group, ordinary acupuncture group, 10 in each group. Short stab group with short stab plus electroacupuncture, general acupuncture group with conventional acupuncture plus electroacupuncture treatment, were selected “inner knee eye ” “calf ” “yinlingquan ” “foot Sanli ”and“ Liangqiu ”acupuncture. Normal group and model group normal crawl, fixed, without intervention. Each 20min, 1 day, 5d for a course of treatment, each treatment interval 2d, a total of 4 courses of treatment. After treatment, the protein expression of DDR2 and type Ⅱ collagen in cartilage was detected by Western Blot (WB); the activities of DDR2, type Ⅱ collagen and MMP13 in cartilage were detected by immunohistochemical method; the expression of DDR2 and type Ⅱ collagen in cartilage was detected by reverse transcription polymerase chain reaction reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the mRNA expression of DDR2 and MMP13. Results: Compared with the normal group, the expression of type Ⅱ collagen and the activity of collagen Ⅱ were significantly decreased (all P <0.01) and the activities of DDR2, MMP13 and mRNA were significantly increased (all P <0.01) (P <0.01), while the activity of DDR2, MMP13 and mRNA expression were significantly decreased (all P <0.01). Compared with the normal acupuncture group Compared with the control group, MMP13, DDR2 activity and mRNA expression were decreased in the short stab group (all P <0.01), and type Ⅱ collagen protein and active protein expression increased significantly (P <0.01). Conclusion: Both electro-acupuncture and electro-acupuncture can promote the repair of articular cartilage in rabbits, and acupuncture plus electro-acupuncture has some advantages. The mechanism may be related to the blockage of type Ⅱ collagen / DDR2 / MMP13 pathway A vicious cycle that inhibits the sustained degradation of extracellular matrix of cartilage.