针刺对干眼的抗炎作用以及与JAK2/STAT3信号转导通路的关系

Anti-inflammatory effects of acupuncture on dry eye and its relationship with the signal pathway of Janus kinase 2/signal transducer and activator of transcription3

目的检测干眼模型兔泪腺JAK2/STAT3信号转导通路中相关因子表达变化,探讨针刺对干眼的抗炎作用以及与JAK2/STAT3信号转导通路的关系。方法选取42只健康新西兰兔,随机分为7组,空白组、模型组、西药组、针刺组、假针刺组、拮抗组、拮抗加针刺组,每组6只。空白组不做处理,其余各组动物每天800、1100、1400和1800时皮下注射氢溴酸东莨菪碱,每次2.0 mg,连续21 d,制作干眼炎症模型。针刺组于造模成功后开始给予针刺治疗,穴位:睛明、攒竹、丝竹空、太阳穴、瞳子髎。其余各组在建模成功的基础上给予相应处理。采用Schirmer试纸检测各组动物不同时间段泪液分泌量,ELISA法检测干眼模型兔泪腺中乙酰胆碱(acetylcholine,ACh)及受体α7nAChR含量的变化,运用实时定量PCR检测泪腺内STAT3、JAK2 mRNA表达情况。结果 Schirmer I实验结果说明氢溴酸东莨菪碱造模21 d有效,西药和针刺治疗可增加干眼模型兔的泪液分泌量。与空白组相比,模型组、假针刺组、拮抗组、拮抗加针刺组干眼模型兔泪腺中α7nAChR含量和ACh含量均显著降低,差异均有统计学意义(均为P<0.05)。与模型组相比,空白组、西药组、针刺组干眼模型兔泪腺中α7nAChR含量和ACh含量均显著升高,差异均有统计学意义(均为P<0.05)。与空白组相比,西药组、针刺组兔泪腺中JAK2 mRNA相对表达量显著升高,差异均有统计学意义(均为P<0.05);拮抗组、拮抗加针刺组兔泪腺中JAK2 mRNA相对表达量显著降低,差异均有统计学意义(均为P<0.05)。与模型组相比,西药组、针刺组兔泪腺中STAT3 mRNA和JAK2 mRNA相对表达量均显著升高,差异均有统计学意义(均为P<0.05);拮抗加针刺组兔泪腺中JAK2 mRNA相对表达量显著降低,差异有统计学意义(P<0.05)。结论针刺能够促进干眼模型兔泪液分泌,可以增加泪腺中α7nAChR和ACh含量,同时增加泪腺内STAT3、JAK2 mRNA表达,针刺治疗干眼的抗炎作用可能与JAK2/STAT3信号通路的激活相关。

Objective To investigate the effect of acupuncture on the related factors in the signal pathway of Janus kinase 2/signal transducer and activator of transcription3(JAK2/STAT3) in the dry-eye model rabbits and explore the mechanism of acupuncture on dry eye.Methods Totally 42 healthy New Zealand white rabbits were randomly divided into seven groups:blank group,model group,western medicine group,acupuncture group,sham acupuncture group,antagonist group,and antagonistic plus acupuncture group,with six rabbits in each group.The rabbits in the blank group were not treated and the rabbits in the other groups were injected subcutaneously with scopolamine hydrobromide at 2.0 mg every time for 21 days at at 8:00 am,14:00 pm and 18:00 pm to prepare dry eye inflammation model rabbits.The rabbits in the acupuncture group began to receive acupuncture treatment after successful modeling.The acupoints were jingming,cuanzhu,sizhukong,taiyang and tongziliao.Rabbits in other groups were given corresponding treatments after successful modeling.Schirmer test strips were used to detect tear secretions of rabbits in different groups at different time points.The contents of acetylcholine(ACh) and its receptor named α7 nAChR in lacrimal glands of dry eye model rabbits were detected by ELISA.The expressions of STAT3 and JAK2 mRNA in the lacrimal glands were detected by real-time quantitative PCR.Results The results of Schirmer I test indicated that scopolamine hydrobromide was effective for 21 days.Western medicine and acupuncture treatments could increase the tear secretions of dry eye model rabbits.Compared with the blank group,the contents of α7 nAChR and ACh in the lacrimal glands of the model group,the sham acupuncture group,the antagonist group and the antagonistic plus acupuncture group were significantly lower,and the differences were statistically significant(all P<0.05).Compared with the model group,the contents of α7 nAChR and ACh in the lacrimal glands of the blank group,western medicine group and acupuncture group were significantly increased,and the differences were statistically significant(all P<0.05).Compared with the blank group,the relative expressions of JAK2 mRNA in the lacrimal glands of western medicine group and acupuncture group were significantly increased,and the differences were statistically significant(both P<0.05) and the relative expressions of JAK2 mRNA in the lacrimal glands of antagonist group and antagonistic plus acupuncture group were significantly decreased,and the differences were statistically significant(both P<0.05).Compared with the model group,the relative expressions of STAT3 mRNA and JAK2 mRNA in the lacrimal glands of the western medicine group and the acupuncture group were significantly increased,and the differences were statistically significant(all P<0.05),and the relative expression of JAK2 mRNA in the lacrimal gland of the antagonistic plus acupuncture group was significantly decreased,and the difference was statistically significant(P<0.05).Conclusion Acupuncture can promote the lacrimal secretions of dry eye model rabbits and increase the contents of α7 nAChR and ACh in the lacrimal glands and enhance the expressions of STAT3 and JAK2 mRNA in the lacrimal glands.The anti-inflammatory effect of acupuncture on dry eye may be related to the activation of JAK2/STAT3 signal pathway.