丹参酮ⅡA对脓毒症急性肺损伤小鼠多糖包被的保护作用

Protective effect of tanshinoneⅡA on glycocalyx in septic mice with acute lung injury

目的探讨丹参酮ⅡA对脓毒症急性肺损伤小鼠多糖包被的作用及机制。方法将8周龄雄性昆明小鼠随机分为3组:假手术组(S组)、脓毒症组(CLP组)及丹参酮ⅡA治疗组(TSN组)。采用盲肠结扎穿孔法建立脓毒症模型;TSN组于术后3h和12h用丹参酮ⅡA(15mg/kg,腹腔内注射)干预,S组和CLP组于同时间点给予等量生理盐水。观察CLP组及TSN组术后7d生存率。于术后24h时收集BALF及肺组织,采用ELISA法测定BALF中炎症因子及多糖包被标志物水平;HE染色观察肺部病理变化;采用比色法检测肺组织MDA含量及SOD活性。结果与S组比较,CLP组BALF中TNF-α及IL-6水平升高(P<0.01),肺损伤评分增加(P<0.01),肺组织MDA含量增加、SOD活性下降(P<0.01),肺组织syndecan-1、heparin sulfate及thrombomodulin水平明显升高(P<0.01);与CLP组比较,TSN组7d生存率升高(P<0.05),BALF中TNF-α和IL-6水平降低(P<0.05),肺损伤评分下降(P<0.05),肺组织MDA含量下降(P<0.01),SOD活性增加(P<0.05),肺组织syndecan-1、heparin sulfate及thrombomodulin水平下降(P<0.01)。结论丹参酮ⅡA可减轻肺脏多糖包被损伤,其机制可能与抑制炎症反应及氧化应激有关。

Objective To investigate effects of tanshinoneⅡA on glycocalyx in mice with acute lung injury induced by sepsis and its underlying mechanisms.Methods Seventy-two eight-week old male kunming mice were randomly divided into a sham group,a septic model group,and an tanshinoneⅡA treatment group.Septic model was established by cecal ligation and puncture(CLP)in mice.Mice in the tanshinoneⅡA treatment group were administered with tanshinoneⅡA(15 mg/kg,intraperitoneally injection,ip)at 3 hand 12 h,while mice in the model and sham group were administered with the same amount of saline.Fifteen mice in CLP group and tanshinoneⅡA treatment group were randomly chosen to observe the 7 dsurvival rate.The other 12 mice were killed at 24 hand the bronchoalveolar lavage fluid(BALF)and lung tissue were collected.ELISA was used to detect the cytokines and degradation markers of glycocalyx in the BALF.Hematoxylin-eosin staining was preformed to show the morphological changes in the lung tissues.Malondialdehyde(MDA)content and the activity of superoxide dismutase(SOD)in the lung tissues were examined by colorimetry.Results Compared with the sham group,the lung morphological damage score,the levels of TNF-αand IL-6 in the BALF,MDA content in the lung,and expression of syndecan-1,heparin sulfate,and thrombomodulin increased greatly in the CLP group(P<0.05 or P<0.01).Compared with the CLP group,the 7 d survival rate and lung SOD activity increased significantly in the tanshinoneⅡA treatment group(P<0.01);the lung morphological damage score,the levels of the cytokines(TNF-αand IL-6),the levels of the degradation markers of glycocalyx(syndecan-1,heparin sulfate,and thrombomodulin)in the BALF and MDA content in the lung decreased obviously in the tanshinoneⅡA treatment group(P<0.05 or P<0.01).Conclusions TanshinoneⅡA could ameliorate the damage of lung glycocalyx,and its mechanism may be related to the inhibition of inflammatory response and oxidative stress.