电针足三里对严重烫伤大鼠凝血功能和血浆促炎性细胞因子的影响

目的:探讨电针“足三里”穴对严重烫伤大鼠凝血功能指标血浆凝血酶原时间(PT)、活化部分凝血活酶时间(APTT)、凝血酶时间(TT)、纤维蛋白原(Fbg)含量和血浆促炎性细胞因子肿瘤坏死因子α(TNF-α)、白介素1β(IL-1β)、白介素6(IL-6)及高迁移率族蛋白B1 (HMGB1)水平的影响.方法:健康雄性Sprague-Dawlay大鼠40只,随机分为5组(每组n=8):假烫伤(NC)组、烫伤(Tem)组、足三里+烫伤(ST36)组、非经非穴+烫伤(NA)组和足三里+烫伤+α-银环蛇毒素(α-BGT)组.建立大鼠30％总体表面积(TBSA)Ⅲ度烫伤模型;“足三里”穴取双侧膝关节外下方腓骨小头下5 mm处;烫伤后立即行(3V,2 ms,3 Hz)脉冲电流持续刺激12 min,每8h刺激1次.烫伤后48 h,经右颈总动脉取血,全自动血凝仪检测血浆PT、APTT、TT和Fbg含量;酶联免疫吸附试验(ELISA)检测血浆TNF-α、IL-1β、IL-6和HMGB1的水平.结果:严重烫伤后48 h,烫伤组大鼠血浆PT、APTT缩短,TT延长,Fbg升高;与烫伤组相比,电针足三里组大鼠PT、APTT延长(P＜0.05),TT缩短(P＜0.05),Fbg含量下降(P＜0.05);同时,烫伤使大鼠血浆IL-1β、IL-6和HMGB1水平明显增高;而TNF-α并未发现明显变化;针刺足三里后血浆IL-1β、IL-6和HMGB1水平均显著下降(P＜0.01);而α-BGT的处理显著逆转针刺足三里这一效应;非经非穴组、α-BGT组PT、APTT、TT和Fbg,血浆促炎性细胞因子IL-1β、IL-6和HMGB1水平与“足三里”组比较差异显著(P＜0.05,P＜0.01).结论:电针足三里穴能明显改善严重烫伤后大鼠的凝血功能状态,降低血浆促炎性细胞因子水平,该效应可能是通过激活含α7亚基N型胆碱能受体介导的胆碱能抗炎通路实现的.

Effects of Electro-Acupuncture Zusanli(ST36) on Coagulation and Pro-inflammatory Cytokines in Severely Thermal Injury Rats

Objective: To investigate the effects of electro-acupuncture zusanli on the plasma prothrombin time(PT),activated partial thromboplastin time(APTT),thrombin time(TT),fibrinogen(Fbg),the plasma tumor necrosis factor-α(TNF-α),interleukin 1β(IL-1β),interleukin 6(IL-6) and high mobility group box-1(HMGB1) protein levels in thermal injury rats.Methods: Male Sprague-Dawley(SD) rats subjected to 30 percent of total body surface area(TBSA) Ⅲ degree thermal injury were employed as the models.Forty rats were randomly divided into five groups: sham thermal injury/normal control(NC) group,scalded/thermal injury(Tem) group,zusanli + thermal injury(ST36) group,non-acupoints + thermal injury(NA) group,and zusanli + thermal injury + α-bungarotoxin(α-BGT) group,with 8 rats in each group.Zusanli points were electro-acupunctured with constant voltage(3 V,2 ms,3 Hz) for 12 minutes(one per 8 hours) immediately after 30% TBSA Ⅲ degree thermal models were established.At the 48th hour after thermal injury,the blood samples were taken via the right common carotid artery in every group,the plasma levels of PT,APTT,TT and Fbg were determined by Sysmex CA6000;the plasma levels of TNF-α,IL-1β,IL-6 and HMGB1 were measured using enzyme-linked immunosorbent assays(ELISA).Results: Thermal injury produced shorter plasma PT and APTT,longer TT and elevated the fibrinogen levels;the plasma concentration of IL-1β,IL-6 and HMGB1 significantly increased after thermal injury.Electrical stimulation of ST36 not only significantly lengthened the plasma PT and APTT,shortened TT and decreased fibrinogen levels(P<0.05,respectively),but also inhibited the plasma pro-inflammatory cytokines IL-1β,IL-6 and HMGB1 production(P<0.01,respectively) as compared with thermal injury controls.However,α-bungarotoxin pre-treatment reversed the electrical stimulation effects(P<0.05,P<0.01,respectively).Conclusion: Electro-acupuncture at zusanli points improved coagulation function,attenuated pro-inflammatory cytokine responses,and the effects might be realized by limiting the inflammatory responses via alpha 7-nicotinic acetylcholine receptors activated cholinergic anti-inflammatory pathway in rats.