推拿退热六法对大肠杆菌内毒素致热幼兔外周环氧合酶2、前列腺素E2表达的影响

Effects of six antipyretic methods of tuina on COX-2/PGE2 expression in peripheral of LPS-induced young rabbits

目的观察推拿退热六法对大肠杆菌内毒素致热幼兔的退热作用及外周作用机制。方法将24只幼兔随机分为盐水对照组、模型组、退热六法组,每组8只。模型组、退热六法组采用耳缘静脉大肠杆菌内毒素注射法制备发热模型。模型建立后60分钟,退热六法组给予退热六法干预:开天门、推坎宫、揉太阳、揉耳后高骨、清天河水、推脊。造模后每20分钟记录体温1次,连续测量温度180分钟。观察各组幼兔体温变化及血常规白细胞计数、中性粒细胞百分比,采用酶联免疫吸附试验(ELISA法)检测幼兔外周血、肝、肺组织前列腺素E2(prostaglandin E2,PGE2)表达水平,采用免疫印迹法(Western Blot法)检测幼兔肝、肺组织环氧合酶2(cyclooxygenase,COX-2)表达水平。结果与盐水对照组相比,模型组幼兔体温升高(P<0.05),退热六法组幼兔体温升高明显低于模型组(P<0.05)。干预后120分钟,与盐水对照组相比,模型组幼兔血白细胞降低、中细粒细胞百分比显著升高(P<0.05)。退热六法组中性粒细胞百分比较模型组降低明显(P<0.05),白细胞降低无统计学意义(P>0.05)。ELISA结果显示,与盐水对照组比较,模型组外周血、肝、肺组织PGE2均显著上升(P<0.05)。退热六法组经干预后,各组织PGE2表达量均较模型组降低(P<0.05),其中外周血PGE2表达量较盐水对照组无统计学意义(P>0.05),肝、肺组织表达较盐水对照组差异显著(P<0.05)。Western Blot结果显示,模型组肝、肺组织COX-2蛋白表达增加,与盐水对照组相比具有统计学差异(P<0.05)。与模型组比较,退热六法组肝、肺组织COX-2表达水平显著下降(P<0.05)。结论推拿退热六法对大肠杆菌内毒素致热幼兔有良好的退热作用,其体温调节机制是通过下调血的中性粒细胞百分比,降低外周血、肝、肺PGE2表达水平,抑制肝、肺COX-2蛋白表达来调节机体抗炎免疫功能实现。

Objective To observe the antipyretic effect and peripheral mechanism of six methods of tuina antipyretic on endotoxin-induced fever in young rabbits.Methods 24 young rabbits were randomly divided into saline control group(n=8),model group(n=8)and antipyretic six methods group(n=8).The fever model was established by endotoxin injection of E.coli from ear vein in model group and antipyretic six methods group.Sixty minutes after the establishment of the model,the six antipyretic methods group was given six-antipyretic interventions:opening Tianmen,pushing Kangong,rubbing Taiyang,rubbing Erhougaogu,clearing the Tianheshui and pushing the spine.After modeling,the body temperature was recorded once every 20 minutes,and the temperature was continuously measured for 180 minutes.The changes of body temperature,white blood cell count and neutrophil percentage of young rabbits in each group were observed.The expression level of PGE2 in peripheral blood,liver and lung of young rabbits was detected by ELISA,and the expression level of COX-2 in liver and lung was detected by Western Blot.Results Compared with saline control group,the body temperature in the model group increased(P<0.05),and the body temperature in the antipyretic six methods group was significantly lowered than that in the model group(P<0.05).120 minutes after intervention,the white blood cells of young rabbits in model group decreased and the percentage of neutrophils increased significantly compared with saline group(P<0.05).Compared with model group,the percentage of neutrophils in antipyretic group decreased significantly(P<0.05).Elisa results showed that compared with saline control group,PGE2 in peripheral blood,liver and lung of model group increased significantly(P<0.05),and the expression of PGE2 in peripheral blood was lower than that of model group(P<0.05).Western blot results showed that the expression of COX-2 in liver and lung of model group increased significantly compared with saline control group(P<0.05).Compared with the model group,the expression level of COX-2 in liver and lung of the six-antipyretic method group decreased significantly(P<0.05).Conclusion The six antipyretic methods of tuina have a good antipyretic effect on endotoxin-induced fever in young rabbits,and its body temperature regulation mechanism is realized by down-regulating the percentage of neutrophils in blood,reducing the expression level of PGE2 in peripheral blood,liver and lung,and inhibiting the expression of COX-2 in liver and lung.