Mechanism of electroacupuncture in acute lung injury induced by endotoxin in mice
KONG Chang,LIU Wei-qiang,LI Meng-han
Abstract:
Objective To investigate the role of cAMP response element-binding protein 3 (CREB3) in alleviating Golgi stress in endotoxin -induced acute lung injury in mice through electroacupuncture(EA). Methods 24 healthy male SPF-grade C57BL/6J mice, aged 7-8 weeks and weighing 25~30 g, were randomly divided into 4 groups (n=6):WT operation group (WT group), endotoxin-induced acute lung injury model group(LPS group), model+electroacupuncture group (EA+LPS group), and model+electroacupuncture at non-acupoint group (SEA +LPS group). The EA +LPS group received electroacupuncture at bilateral Zusanli and Feishu acupoints for 5 days prior to model induction, while the SEA+LPS group was stimulated at distant non-acupoints using the same method. The LPS group, EA+LPS group, and SEA+LPS group received intraperitoneal injections of lipopolysaccharide (LPS) at a dose of 12 mg/kg, while the WT group was given an equivalent volume of normal saline. 12 hours after the injection of LPS or normal saline, blood samples were collected, and serum was separated to measure the concentrations of tumor necrosis factor (TNF)-a, Interleukin (IL)-1茁, and IL-6. Lung tissues were collected for pathological injury scoring, immunofluorescence staining to observe Golgi morphology,and detection of CREB3 protein expression levels, as well as the protein expression of Golgi stress-related markers Golgin97, GRASP65, and GOLPH3. Results Compared with the LPS group, the EA+LPS group showed a decrease in lung injury scores,reduced Golgi damage under the microscope, and lower concentrations of serum TNF-a, serum IL-1,and serum IL-6 (P<0.05). Additionally, the expression levels of CREB3 protein in lung tissue were downregulated, while the expression of Golgi stress-related proteins Golgin97 and GRASP65 were upregulated,and GOLPH3 expression was downregulated (P<0.05). However, in the SEA +LPS group, there were no statistically significant differences in these indicators (P>0.05). CREB3 knockout mice can reverse the therapeutic effects of electroacupuncture on Golgi apparatus stress and sepsis induced lung injury. Conclusion The mechanism by which CREB3 mediates the alleviation of endotoxin -induced acute lung injury in mice through electroacupuncture at Zusanli and Feishu acupoints may be related to Golgi stress.
