电针调控CD73及腺苷信号对脓毒症急性肾损伤的影响
Effect of Eletroacupuncture on CD73 and Adenosine Signaling in Sepsis-Associated Acute Kidney Injury
程辞云 1杨玲玲 1胡秀敏 1陈莎莎 1王蕊 2余常1
作者信息
- 1. 成都中医药大学针灸推拿学院,四川成都 611137
- 2. 四川中医药高等专科学校针灸推拿学院,四川绵阳 621000
- 折叠
摘要
目的:观察电针对小鼠脓毒症急性肾损伤的影响,明确电针改善脓毒症急性肾损伤与调控CD73及下游腺苷信号之间的联系.方法:将C57BL/6J小鼠随机分为空白组(Control)、模型组(LPS)、电针组(EA)、假电针组(shamEA);采用脂多糖(Lipopolysaccharide,LPS)腹腔注射建立脓毒症急性肾损伤小鼠模型;在造模前及造模后12 h,给予小鼠"足三里"穴10 Hz、3.0 mA、30 min的电针干预,假电针干预仍连接电针仪但不通电;采用酶联免疫吸附法(Enzyme Linked Immunosorbent Assay,ELISA)检测肾组织肿瘤坏死因子-α(Tumor Necrosis Factor Alpha,TNF-α)水平变化;采用免疫印记法(Western Blot,WB)检测肾组织 IL-6、CD73、A2AR、HIF-1α蛋白表达的变化.结果:与Control组比较,LPS组小鼠肾组织中TNF-α水平、IL-6蛋白表达升高(P<0.001);与LPS组比较,EA组小鼠肾组织中TNF-α水平、IL-6蛋白表达降低(P<0.001).与Control组比较,LPS组小鼠肾组织中CD73、A2A R蛋白表达减少(P<0.001)、HIF-1α蛋白表达上升(P<0.001);与LPS组比较,EA组小鼠肾组织中CD73、A2AR蛋白表达增强(P<0.001)、HIF-1 α蛋白表达下降(P<0.001).结论:电针可降低脓毒症急性肾损伤小鼠肾组织中IL-6蛋白表达,降低TNF-α水平,具有抗炎效应;电针减轻脓毒症急性肾损伤可能与其调控CD73及下游腺苷信号相关.
Abstract
Objective:To observe the effect of electroacupuncture on sepsis-associated acute kidney injury(SA-AKI)and clarify the link between electroacupuncture(EA)to improve SA-AKI and the regulation of CD73 and adenosine signaling.Methods:C57BL/6J mice were randomly divided into Control,LPS,EA and shamEA group.The murine model of acute kidney injury was established by an intraperitoneal injection of LPS.EA stimulation was performed 12 hours before and after intraperitoneal injection of LPS and applied at the bilateral Zusanli(足三里,ST36)acupoint,with a parameter of 10 Hz and 3.0 mA for an amount of time equaling 30 minutes each.The content of the Tumor Necrosis Factor Alpha(TNF-α)in kidney tissue was quantified by the Enzyme-Linked Immunosorbent Assay(ELISA)kits.Meanwhile,the protein expression of IL-6,CD73,A2AR,and HIF-1α in kidney tissue was extracted from the Western Blot(WB).Results:Compared with Control group,the content of TNF-α and the protein expression of IL-6 in kidney tissue were significantly increased in LPS group(P<0.001).In contrast,the EA group has a significant reduction in the level of TNF-α and protein expression of IL-6 compared with the LPS group(P<0.001).There was a significantly low protein expression of CD73 and A2AR in LPS group(P<0.001),in comparison to the Control group,whereas that of HIF-1α was significantly higher.While in EA group,the expression of CD73 and A2AR increased dramatically(P<0.001),but HIF-1α protein expression decreased significantly compared to the LPS group(P<0.001).Conclusion:Electroacupuncture,hence,through downregulation of IL-6 protein expression and TNF-α content in kidney tissues of the mouse model with SA-AKI,exerts anti-inflammatory effects.The efficacy of electroacupunc-ture to ameliorate SA-AKI may be associated with modulation of CD73 and downstream adenosine signaling.
关键词
电针/抗炎效应/脓毒症急性肾损伤/腺苷/胞外-5'-核苷酸酶/腺苷信号A2A受体Key words
Electroacupuncture/Anti-inflammatory effect/Sepsis-associated acute kidney injury/Adenosine/CD73/A2AR引用本文复制引用
基金项目
四川省中医药局中医药科研专项(2021MS103)
四川省中医药局中医药科研专项(2021MS231)
四川省自然科学基金(2022NSFSC0852)
出版年
2024