The mechanism of 5-hydroxytryptamine in electroacupuncture pre-treatment for myocardial injury in sepsis mice
10.3760/cma.j.cn121430-20231220-01098
- VernacularTitle:5-羟色胺在电针预处理脓毒症小鼠心肌损伤中的作用机制研究
- Author:
Yunfeng LI
1
;
Luyao QI
;
Bingqing OUYANG
;
Huan GENG
;
Xiaoxiao LI
;
Ming LEI
Author Information
1. 上海中医药大学附属第七人民医院重症医学科,上海 201203
- Keywords:
Sepsis;
Electroacupuncture;
Zusanli;
Myocardial injury;
MB isoenzyme of creatine kinase;
Mice;
Glycolysis
- From:
Chinese Critical Care Medicine
2024;36(8):835-840
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To observe the changes of 5-hydroxytryptamine (5-HT) level in myocardial tissue of pre-treatment mice with sepsis myocardial injury by electroacupuncture at Zusanli point, and to explore the protective effect and possible mechanism of electroacupuncture on myocardial injury in sepsis.Methods:Twenty male C57BL/6 mice were divided into control group (NC group), sepsis model group (LPS group), electroacupuncture group (EA group) and electroacupuncture + fluoxetine group (EA+FLU group) by random number table method, with 5 mice in each group. The myocardial injury model of sepsis was established by intraperitoneal injection of lipopolysaccharide (LPS) 10 g/L. The NC group was intraperitoneally injected with the same amount of normal saline. 3 days before mold making, EA group and EA+FLU group were electrocuted at Zusanli point on both sides for 15 minutes, once a day for 3 days. The EA+FLU group was intraperitoneally injected fluoxetine 1.4 g/L before electroacupuncture. After modeling, the cardiac histopathological changes were observed by hematoxylin-eosin (HE) staining. The serum levels of inflammatory cytokines interleukins (IL-6, IL-8), and tumor necrosis factor-α (TNF-α), the content of 5-HT in myocardial tissue, myocardial injury markers MB isoenzyme of creatine kinase (CK-MB), and cardiac troponin I (cTnI), and the levels of adenosine triphosphate (ATP) and lactic acid in myocardial tissue were detected. Quantitative polymerase chain reaction (qPCR) was used to detect the mRNA expressions of 5-hydroxytryptamine transporter (5-HTT), hexokinase 2 (HK2) and glucose transporter 4 (GLUT4) in myocardial tissue. GLUT4 expression in myocardial tissue was detected by immunofluorescence assay.Results:Compared with NC group, the serum levels of IL-6, IL-1β and TNF-α, myocardial 5-HT content, myocardial tissue injury markers CK-MB, cTnI in LPS group and EA+FLU group were significantly increased. Compared with LPS group, the above indexes in EA group were significantly decreased [IL-6 (ng/L): 443.03±156.16 vs. 19?843.75±0.00, IL-1β (ng/L): 75.72±10.60 vs. 894.66±350.88, TNF-α (ng/L): 46.17±4.71 vs. 533.01±170.58, 5-HT (μg/L): 161.19±5.96 vs. 244.74±14.38, CK-MB (ng/L): 468.21±12.46 vs. 662.02±22.54, cTnI (ng/L): 0.83±0.05 vs. 0.99±0.08, all P < 0.05]. Compared with NC group, the levels of ATP in myocardium of LPS group, EA group and EA+FLU group were significantly decreased, the levels of lactic acid in myocardium were significantly increased. Compared with LPS group, the level of ATP in myocardium of EA group was significantly increased, the level of lactic acid in myocardium was significantly decreased [ATP (mmol/L): 0.10±0.01 vs. 0.08±0.01, lactic acid (mmol/L): 56.03±1.07 vs. 72.45±4.32, both P < 0.05]. Compared with NC group, the mRNA expression of HK2 in myocardium of LPS group was significantly increased, and the mRNA expressions of GLUT4 and 5-HTT were significantly decreased. Compared with LPS group, the mRNA expression of HK2 in myocardium of EA group was significantly decreased, the mRNA expressions of GLUT4 and 5-HTT were significantly increased [HK2 mRNA (relative expression level): 0.73±0.19 vs. 1.82±0.57, GLUT4 mRNA (relative expression level): 1.00±0.33 vs. 0.47±0.18, 5-HTT mRNA (relative expression level): 1.18±0.31 vs. 0.38±0.15, all P < 0.05]. Compared with NC group, the fluorescence intensity of GLUT4 in LPS group and EA+FLU group were significantly decreased. Compared with LPS group, the fluorescence intensity of GLUT4 in EA group was significantly enhanced. Conclusions:Electroacupunctureat Zusanli can reduce the content of 5-HT in myocardial tissue of sepsis mice, and its regulatory mechanism may be related to the regulation of 5-HTT and GLUT4.