基于AMPK能量调节功能探讨中医实热“上火”的发病机制  

Discussion on pathogenesis of excessive heat 'shanghuo' based on the energy regulating function of AMPK

作　　者：包洁[1] 张喜召 窦晓兵[1] 范永升[1] BAO Jie;ZHANG Xi-zhao;DOU Xiao-bing;FAN Yong-sheng(Zhejiang Chinese Medical University,Hangzhou 310053,China)

机构地区：[1]浙江中医药大学,杭州310053

出　　处：《中华中医药杂志》2018年第9期4171-4176,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家重点基础研究发展计划(973计划)项目(No.2014CB543001),浙江省自然科学基金青年基金项目(No.LQl8H270004)

摘　　要：目的:以腺苷酸活化蛋白激酶(AMPK)为核心探讨实热"上火"人群体内能量代谢的变化,为阐明实热"上火"的发病机制提供参考依据。方法:磷钼酸比色法检测外周血中三磷酸腺苷(ATP)含量,RTPCR法检测外周血白细胞中AMPK、信号传导及转录激活因子3(STAT3)mRNA含量,Western blot法检测其蛋白表达水平,试剂盒检测细胞耗氧率(OCAR),ELISA法测定血浆中白介素(IL)-6、IL-17、肿瘤坏死因子α(TNF-α)等细胞因子含量。结果:与正常对照组人群比较,实热"上火"组人群外周血中ATP含量显著上升(P<0.01),外周血白细胞中AMPK蛋白的磷酸化水平显著下降(P<0.05),OCAR也显著下降(P<0.05);外周血白细胞中STAT3蛋白磷酸化水平下降(P<0.05),血浆中IL-6含量有所上升,而IL-17、TNF-α则下降(P<0.05)。结论:实热"上火"状态下,体内能量物质ATP显著增多,AMPK功能受到抑制,引起机体的正反馈调节,出现合成代谢减少,OCAR下降,分解代谢亢进的状态。同时,受AMPK活性下降的影响,体内STAT3活性也随之减弱,炎性细胞因子比例下调,出现反馈性的炎症免疫抑制反应。Objective： At the core of AMP-activated protein kinase （AMPK）, the changes of energy metabolism in the body of excessive heat were discussed in order to provide reference for the pathogenesis of ‘shanghuo＇. Methods： The content of adenosine triphosphate （ATP） was detected by the method of phosphomolybdic acid colorimetry. AMPK and STAT3 mRNA was detected by the method of RT-PCR and protein content by the method of Western blot. Peripheral blood leucocytes oxygen consumption rate （OCAR） was detected by corresponding kit. The content of IL-6, IL-17 and TNF-α in the plasma were measured by the method of ELISA. Results： The content of ATP in the people with excessive heat ‘shanghuo＇ was increased observably （P〈0.01）. The mRNA content of AMPK in peripheral blood leucocytes was increased similarly （P〈0.05）, while the phosphorylation of protein decreased significantly （P〈0.05）, and the rate of cell oxygen consumption also decreased significantly. On the other hand, the mRNA and protein phosphorylation of STAT3 showed tendency to ascend. As for inflammatory cytokines in the plasma, the content of IL-6 increased, and IL-17 and TNF-α decreased significantly （P〈0.05）. Conclusion： In the state of excessive heat ‘shanghuo＇, the energy of human body increased and the function of AMPK is restrained, causing the positive feedback regulation. The state of decreased anabolism, decreased oxygen consumption rate and hyper catabolism are found. Meanwhile the activity of STAT3 is impaired and inflammatory cytokines are decreased, leading to a state of immunosuppression.

关 键 词：实热“上火” 发病机制 腺苷酸活化蛋白激酶 三磷酸腺苷 信号传导及转录激活因子3 炎症 反应 

分 类 号：R254[医药卫生—中医内科学]