丹酚酸B对脓毒症小鼠血管平滑肌细胞炎症反应的影响:circACTA2在其中的作用  

Effect of salvianolic acid B on inflammatory responses of vascular smooth muscle cells in septic mice:role of circACTA2

作　　者：张曼莉[1] 赵景茹[2] 尹化 张曼娜 宋璇 佟飞[1] Zhang Manli;Zhao Jingru;Yin Hua;Zhang Manna;Song Xuan;Tong Fei(Department of Critical Care Medicine,The Second Hospital of Hebei Medical University,Shijiazhuang 050000,China;Department of Neurology,Hebei General Hospital,Shijiazhuang 050051,China;Department of Clinical Laboratory,The Second Hospital of Hebei Medical University,Shijiazhuang 050000,China)

机构地区：[1]河北医科大学第二医院重症医学科,石家庄050000 [2]河北省人民医院神经内科,石家庄050051 [3]河北医科大学第二医院检验科,石家庄050000

出　　处：《中华麻醉学杂志》2024年第2期225-231,共7页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金项目青年科学基金(82001229);河北省自然科学基金中医药联合基金培育项目(H2023206276);河北省政府资助临床医学优秀人才培养项目(ZF2023113)。

摘　　要：目的评价丹酚酸B(Sal B)对脓毒症小鼠血管平滑肌细胞(VSMC)炎症反应的影响及circACTA2在其中的作用。方法在体实验:健康雄性C57BL/6小鼠81只,6~8周龄,采用随机数字表法分为3组(n=27):假手术组、脓毒症组和Sal B组。采用盲肠结扎穿孔法制作脓毒症模型。模型制备成功后,Sal B组腹腔注射Sal B 7 mg/kg,1次/d,连续2 d。每组随机取20只小鼠用于造模后7 d内测定SBP、DBP、MAP、全血乳酸(Lac)浓度,记录生存情况。造模后48 h时每组随机取7只小鼠,取动脉血管组织,采用免疫荧光染色法测定IL-1β表达,分别采用Western blot法和qRT-PCR法测定IL-1β、TNF-α、IL-6及其mRNA的表达,采用qRT-PCR法测定circACTA2的表达。细胞实验:小鼠动脉VSMC培养后,采用随机数字表法分为6组(n=3):对照组(C组)、LPS组、Sal B组、si-circACTA2+C组、si-circACTA2+LPS组和si-circACTA2+Sal B组。LPS组采用LPS(终浓度1 μg/ml)、Sal B组采用LPS(终浓度1 μg/ml)和Sal B (终浓度5 μmol/L)孵育24 h。si-circACTA2+C组仅用si-circACTA2转染VSMC。si-circACTA2转染VSMC 24 h后,si-circACTA2+LPS组用LPS (终浓度1 μg/ml)、si-circACTA2+Sal B组采用LPS(终浓度1 μg/ml)和Sal B (终浓度5 μmol/L)孵育24 h。分别采用Western blot法和qRT-PCR法测定IL-1β、TNF-α、IL-6及其mRNA的表达,采用qRT-PCR法测定circACTA2的表达。结果在体实验:与假手术组相比,脓毒症组SBP、DBP及MAP降低,全血Lac浓度升高,7 d生存率降低,动脉血管组织IL-1β、TNF-α和IL-6及其mRNA表达上调,circACTA2表达下调(P<0.05),IL-1β荧光增强。与脓毒症组相比,Sal B组SBP、DBP及MAP升高,全血Lac浓度降低,7 d生存率升高,动脉血管组织IL-1β、TNF-α和IL-6及其mRNA表达下调,circACTA2表达上调(P<0.05),IL-1β荧光减弱。细胞实验:与C组相比,LPS组IL-1β、TNF-α和IL-6及其mRNA表达上调,circACTA2表达下调(P<0.05)。与LPS组相比,Sal B组IL-1β、TNF-α和IL-6及其mRNA表达下调,circACTA2表达上调(P<0.05)。Objective To evaluate the effect of Salvianolic acid B(Sal B)on the inflammatory responses of vascular smooth muscle cells(VSMCs)in septic mice and the role of circACTA2.Methods In vivo experiment Eighty-one healthy male C57BL/6 mice,aged 6-8 weeks,were divided into 3 groups(n=27 each)by a random number table method:sham operation group,sepsis group and Sal B group.Sepsis model was developed by cecal ligation and puncture.After sucessful preparation of the model,Sal B 7 mg/kg/d was intraperitoneally injected once a day for 2 consecutive days in Sal B group.Twenty mice in each group were randomly selected to measure systolic blood pressure(SBP),diastolic blood pressure(DBP),mean arterial pressure(MAP)and whole blood lactic acid(Lac)and to record the survival within 7 days after developing the model.Seven mice in each group were randomly selected at 48 h after developing the model,and the arterial vascular tissues were collected for determination of the expression of interleukin-1beta(IL-1β)(by immunofluorescence staining),expression of IL-1β,tumor necrosis factor-alpha(TNF-α)and IL-6 protein and mRNA(by Western blot and quantitative real-time polymerase chain reaction,respectively),and expression of circACTA2(by quantitative real-time polymerase chain reaction).Cell experiment Mouse VSMCs were cultured and divided into 6 groups(n=3 each)by a random number table method:control group(C group),lipopolysaccharide(LPS)group,Sal B group,si-circACTA2+C group,si-circACTA2+LPS group,and si-circACTA2+Sal B group.The cells were incubated for 24 h with LPS(final concentration 1μg/ml)in LPS group and with LPS(final concentration 1μg/ml)and Sal B(final concentration 5μmol/L)in Sal B group.VSMCs were transfected with si-circACTA2 only in si-circACTA2+C group.At 24 h after transfection of si-circACTA2 into VSMCs,the cells were incubated with LPS(final concentration 1μg/ml)in si-circACTA2+LPS group and with LPS(final concentration 1μg/ml)and Sal B(final concentration 5μmol/L)for 24 h in si-circACTA2+Sal B group.The expres

关 键 词：丹参酸B RNA 环状 脓毒症 肌细胞 平滑肌 炎症 

分 类 号：R459.7[医药卫生—急诊医学]