择时针刺对炎性疼痛模型P2X7-/-小鼠穴区成纤维细胞p38MAPK表达的影响

Effect of Chrono-acupuncture on the Expression of p38MAPK in Fibroblasts in Acupoint Area of P2X7-/-mice with Inflammatory Pain

目的明确P2X7受体是否通过p38MAPK信号通路介导择时针刺对成纤维细胞骨架的重构。方法选择P2X7-/-小鼠18只,根据辐射热甩尾法所测痛阈值完全随机分作空白组6只、造模组12只,12只造模动物再通过右足注射完全弗氏佐剂建立炎性疼痛模型。模型成功建立后,根据各动物所测痛阈值随机分为模型组6只、模型+针刺组6只。模针组动物捆绑固定后,在特定时间点ZT8(15:00)针刺足三里穴,每5 min捻转行针1次,每次行针1 min,频率为120次/min。每天治疗1次,每次留针30 min,连续治疗7天。空白组和模型组在同一时间点进行相同方法的捆绑固定30 min,不针刺。采用Western Blot和RT-PCR检测小鼠穴区p38MAPK蛋白表达量和p38MAPK基因表达。结果模型组小鼠皮肤组织中p38MAPK蛋白表达与空白组相比显著升高,有统计学意义(P<0.01);模针组小鼠皮肤组织中p38MAPK蛋白表达与模型组相比存在降低,有统计学意义(P<0.05)。模型组小鼠穴位皮肤及皮下组织中p38MAPK基因表达,与空白组相比升高,有统计学意义(P<0.05);模针组小鼠穴位皮肤及皮下组织中p38MAPK基因表达与模型组相比降低,有统计学意义(P<0.05)。结论P2X7受体可能并非通过对p38MAPK的调整实现择时针刺对穴区成纤维细胞骨架重构的作用。

Objective To clarify whether P2 X7 receptors mediate the remodeling of the fibroblast cytoskeleton by chrono-acupuncture through the p38 MAPK signaling pathway.Methods The selected 18 P2 X7-/-mice were randomly divided into control group(n=6)and model group(n=12),the model group established an inflammatory pain model by injecting Complete Freund’s Adjuvant(CFA)into the animal’s right foot.After the model was successfully established,according to the measured pain threshold of each animal,they were randomly divided into 6 in the model group and 6 in the model+acupuncture group.After the animals of the acupuncture group were bundled processing,we acupunctured Zusanli Point(ST36)at a specific time point ZT8(15:00).Twisted the needle once every 5 mins,each time lasting 1 min,the frequency is 120 times/min,and the needle is left for 30 mins.Once a day for 7 consecutive days.The model control group and the control group were bundled processing in the same manner for 30 mins at the same time point without acupuncture.At last,the expression of p38 MAPK protein and p38 MAPK gene were detected by Western Blot and RTPCR.Results The expression of p38 MAPK protein in the skin tissue of mice in the model group was significantly higher than that in the control group,with statistical significance(P<0.01).Skin mold needles p38 MAPK protein in mice,and reduced compared to the model group was statistically significant(P<0.05).Compared with the blank group,the expression of p38 MAPK gene in the acupoint skin and subcutaneous tissue of the model control group was increased(P<0.05).Compared with the model control group,the p38 MAPK gene in the acupoint skin and subcutaneous tissue of the mice in the acupuncture group the expression was decreased(P<0.05).Conclusion The P2 X7 receptor may not achieve the effect of chrono-acupuncture on fibroblast cytoskeletal remodeling in the cavity area through the adjustment of p38 MAPK.