Acute lung injury (ALI) is a respiratory disorder characterized by severe inflammation of the alveoli and lung parenchyma. Tetramethylpyrazine (TMP), the main active compound in Ligusticum chuanxiong Hort (LC), can protect against lipopolysaccharide (LPS)-induced ALI. Our study aimed to investigate how TMP protects the endothelial cell barrier in pulmonary capillaries. We administered TMP intraperitoneally at different doses and found that acute lung injury in mice was improved, but not in a dose-dependent manner. TMP toxicity was tested in vitro. We observed that LPS-induced cytoskeletal remodeling was inhibited by TMP. Murine ALI was induced as follows: For the 1st hit, LPS (2 mg/kg) was injected intraperitoneally; after 16 h, for the 2nd hit, LPS (4 mg/kg) was instilled intratracheally. The mice in treatment groups had TMP or dexamethasone administered intraperitoneally 30 min prior to the 1st hit and 30 min past the 2nd hit. Mice were euthanized 24 h after the last injecting. We measured protein and mRNA levels using enzyme-linked immunosorbent assay (ELISA) and reverse transcriptase real-time PCR (RT-qPCR), respectively. The ultrastructural analysis was performed with transmission electron microscopy (TEM) and the cytoskeleton was observed by immunofluorescence. Immunohistochemistry and Western blotting were used to detect protein expression in the Rac1/LIMK1/ZO-1/occludin signal pathway. The results showed that TMP treatment decreased inflammatory cell infiltration and alleviated LPS-induced damage in lung tissue. Also, TMP significantly inhibited the Rac1/LIMK1/ZO-1/occludin signaling pathway. Our findings show that using TMP during sepsis can protect the pulmonary microvascular endothelial cell barrier and suppress inflammation. Therefore, TMP may have a promising therapeutic role in preventing acute lung injury from sepsis.

目的 观察川芎嗪对脂多糖诱导的小鼠ALI血管通透性的保护作用并探讨其可能分子机制.方法 C57BL/6 小鼠(n =50)随机分为5 组:CON组(n =10)、ALI组(n =10)、TMP低(50 mg/kg,n =10),中(100 mg/kg,n =10),高(150 mg/kg,n =10)浓度干预组.ELISA检测IL-1β及TNF-α水平;HE染色观察肺部病理变化;湿干比检测肺水肿程度;BALF白细胞计数、总蛋白量测定及EB实验检测肺血管通透性;WB检测P-mTOR、RhoA蛋白水平;电镜下观察内皮细胞间紧密连接.结果 ALI组IL-1β、TNF-α水平较于CON组升高(P均<0.05);中浓度TMP组IL-1β、TNF-α水平下降明显(P均<0.05),低浓度和高浓度TMP组无显著变化(P均>0.05).HE结果提示TMP中浓度可以显著减缓肺损伤的病理变化.ALI组湿干比高于CON组(P<0.01),TMP治疗可降低湿干比(P<0.05).TMP可降低BALF白细胞计数(P<0.01)、蛋白浓度(P<0.01)及Evans Blue渗漏(P<0.05).ALI组P-mTOR(P<0.01)及RhoA(P<0.01)蛋白表达量高于CON组.TMP组P-mTOR(P<0.01)及RhoA蛋白表达量下降(P<0.05).ALI组紧密连接受损,TMP治疗后紧密连接断裂减轻.结论 TMP可通过活化mTOR/RhoA通路改善肺血管通透性从而对LPS引起的小鼠ALI起到保护作用.

该文旨在从Ras相关的C3肉毒素底物1（Ras-related C3 botulinum toxin substrate 1,Rac1）/LIM激酶1（LIM kinase 1,LIMK1）通路探讨川芎嗪（tetramethylpyrazine,TMP）对脂多糖（lipopolysaccharide,LPS）诱导的人脐静脉内皮细胞（human umbilical vein endothelial cells,HUVECs）骨架重构的保护作用。采用Ⅷ因子免疫荧光鉴定原代内皮细胞;分别用0、50、100、200、300 ng/mL的LPS处理对数生长期的原代HUVECs;采用低、中、高3个剂量的TMP对HUVECs预处理12 h;利用F-actin免疫荧光染色观察细胞骨架变化;通过跨膜电阻值（transepithelial electrical resistance,TEER）和FITC-葡聚糖Transwell实验检测人脐静脉内皮细胞通透性;Western blot检测Rac1、LIMK1的表达以及相应磷酸化水平;免疫荧光观察磷酸化LIMK1蛋白的细胞定位和表达水平变化;实时荧光定量PCR（quantitative real-time PCR,qPCR）检测Rac1和LIMK1的mRNA水平。结果显示,Ⅷ因子免疫荧光鉴定原代提取的内皮细胞符合要求;HUVECs经LPS诱导后,镜下形态由卵圆形向长梭形改变,纤维状肌动蛋白（filament actin,F-actin）增多增粗,TEER降低及单层内皮细胞通透性增加;Rac1、LIMK1的表达及磷酸化水平呈LPS梯度依赖性升高（P＜0.05）;低浓度的TMP降低LPS诱导的F-actin增多增粗作用,并降低Rac1、LIMK1的表达及其磷酸化水平（P＜0.05）;高浓度的TMP可以抑制Rac1激活（P＜0.05）,但对LPS诱导的HUVECs细胞骨架重构和通透性增加没有保护作用（P＞0.05）。该研究结果表明,TMP通过Rac1/LIMK1通路保护LPS诱导的HUVECs骨架重构,抑制由LPS诱导的HUVECs通透性增大,对临床运用TMP改善由内皮细胞高通透性引起的急性肺损伤提供理论依据。

目的 观察川芎嗪(Tetramethylpyrazine,TMP)对细菌脂多糖(LPS)诱导的脐静脉内皮细胞屏障损伤(HUVEC)的保护作用,并探索可能的作用机制.方法 将鉴定好的人脐带静脉内皮细胞根据加药处理因素不同,将实验分为空白对照组、LPS组,LPS+TMP组.对三组进行免疫荧光细胞骨架染色及细胞通透性实验、跨内皮电阻实验.应用Westernblot及qPCR方法进行mTOR表达量的定量分析.结果 免疫荧光细胞骨架染色表明与空白组相比,LPS组应力纤维明显增多、增粗.而应用TMP治疗后应力纤维变少、变细.跨内皮电阻实验表明,加药刺激前三组电阻值差无统计学意义(P＞0.05).加入药物刺激后,LPS组较空白组,电阻值明显降低,而TMP组较LPS组明显增加(P＜0.05).细胞通透性实验表明加药刺激后,LPS组较空白组,通透系数明显增加,而TMP组较LPS组明显降低(P＜0.05).Westernblot及qPCR方法表明LPS组较空白组mTOR表达量增加,而TMP治疗之后mTOR表达量较LPS组下降.结论 TMP可通过调控mTOR的表达量对LPS诱导的HUVEC内皮细胞屏障破坏起到保护作用.

急性肺损伤( acute lung injury,ALI)是一种毁灭性的呼吸系统疾病,导致内皮细胞障碍和细胞骨架改变,从而进展为肺水肿、呼吸衰竭及多器官功能障碍综合征 ( multiple organ dysfunction syndrome, MODS) ,最终导致死亡的发生[1].

目的 探讨慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)患者肺功能对单孔胸腔镜肺部手术相关指标的影响,并评估中重度COPD应用单孔胸腔镜进行肺手术的安全性.方法 回顾性分析上海市肺科医院胸外科2020年1-9月1030例COPD患者行单孔胸腔镜的临床资料.根据慢性阻塞性肺疾病全球倡议解读(GOLD)分级标准将肺功能分为轻度、中重度两组,比较不同程度下患者术中失血量(intraoperative blood loss,IBL)、麻醉时间(anesthesia time,AT)、手术时间(operation time,OT)及术后并发症(postoperative complications,PC)、术后住院天数(postoperative hospital stay days,POD).结果 所有患者均成功完成手术,安全出院.轻度组与中重度组在术中IBL、AT、OT、POD、PC等指标上差异无统计学意义(P>0.05).轻度组与中重度组患者术后并发症的差异无统计学意义.结论 中重度COPD患者较轻度COPD患者术中、术后相关指标改变不明显,应用单孔胸腔镜行肺部手术是安全、可行的.