针灸透刺鹤顶与内膝眼穴对兔膝关节炎软骨细胞免疫微环境调控的影响

Effects of acupuncture and moxibustion at Heding and Neixiyan points on immune microenvironment regulation of chondrocyte in rabbit knee arthritis

目的:探讨透刺鹤顶与内膝眼穴对兔膝关节炎软骨细胞免疫微环境的可能作用机制。方法:将24只新西兰兔随机分为A、B、C三组,对B、C组所有大白兔右侧膝关节腔内注射2%木瓜蛋白酶和0.03 mol/L L-半胱氨酸混合液配制,建立KOA模型。A组为正常对照组给予生理盐水灌胃,灌胃量为2 ml/100g,1次/d,持续4周;B组为造模组,给予生理盐水灌胃,灌胃量为2 ml/100g,1次/d,持续4周;C组给予针灸透刺鹤顶至内膝眼穴位1次/d,每次20 min,持续4周。用ELISA检测血清IL-6表达水平,用Western blot检测关节软骨中TLR4、MyD88、IRF-7、MMP-13表达水平,采用改良Lequesne MG评估方法对各组家兔膝关节的疼痛、步态、关节活动度及关节肿胀等行为学指标,进行计分评价。结果:透刺治疗组IL-6为(0.016±0.007)ng/L与模型组(0.054±0.010)ng/L比较有统计学差异(P<0.05),在关节组织免疫学TLR4、MyD88、IRF-7、MMP-13检测中治疗组与模型组比较均有统计学差异(P<0.05),行为力学比较显示模型组四项各评分明显上升,与空白组和透刺治疗组比较均有统计学差异(P<0.05)。结论:针灸透刺鹤顶与内膝眼穴可以起到调节膝关节免疫微环境的功效,从而起到改善膝关节症状和治疗的作用。

Objective:To explore the possible mechanism of Heding and Neixiyan points on chondrocyte immune microenvironment in rabbits with knee arthritis. Methods:24 New Zealand rabbits were randomly divided into groups A,B and C. KOA model was established by injecting 2%papain and 0.03 mol/L-cysteine mixture into the right knee joint cavity of all rabbits in groups B and C. Group A was given saline for 4 weeks. Group B was given saline for 4 weeks. Group B was given saline for 4 weeks. Group C was given acupuncture and moxibustion once a day from Heding to Neixiyan acupoints for 20 minutes. The expression levels of IL-6 in serum were detected by ELISA,TLR4,MyD88,IRF-7 and MMP-13 in articular cartilage were detected by Western blot,and the pain,gait,range of motion and swelling of knee joints were evaluated by modified Lequesne MG method.Results:Serum IL-6 in penetrating needling treatment group was(0.016±0.007)ng/L and(0.054±0.010)ng/L in model group(P<0.05). There were significant differences in TLR4,MyD88,IRF-7 and MMP-13 between model group and treatment group(P<0.05). Behavioral mechanics comparison showed that the four scores in model group increased significantly compared with blank group and penetrating needling treatment group(P<0.05). There were significant differences between the two groups(P<0.05).Conclusion:Acupuncture and moxibustion through Heding and Neixiyan points can regulate the immune microenvironment of knee joint,thus improving knee joint symptoms and treatment,which is worthy of clinical application.