南通大学学报(医学版)

目的：探讨丹酚酸B(salvianolic acid B, SAB)对脂多糖(lipopolysaccharide, LPS)诱导急性肺损伤(acute lung injury, ALI)的影响。方法：体内采用LPS诱导小鼠模型，分为5组：空白组、LPS组(10 mg/kg)、LPS+地塞米松(dexamethasone,Dex)组[LPS+Dex(2 mg/kg)]、LPS+SAB低剂量组(25 mg/kg)、LPS+SAB高剂量组(50 mg/kg)。体外采用LPS诱导小鼠肺上皮细胞MLE-12细胞模型，分为空白组、LPS组(50 mmol/L)、LPS+Dex组(1μmol/L)、LPS+SAB低剂量组(10μmol/L)、LPS+SAB高剂量组(20μmol/L)。末次给药24 h后检测各组小鼠血常规指标[WBC、中性粒细胞(neutrophils, NEU)、淋巴细胞(lymphocytes, LYM)]，肺功能指标[肺总阻力(lung resistance, RL)、肺出气阻力(pulmonary expiratory resistance, Re)、肺动态顺应性(lung dynamic compliance, Cydn)、呼气峰值流速(peak expiratory flow rate, PEF)和最大通气量(maximum ventilation volume, MVV)]，检测小鼠肺湿干比重(wet dry ratio, W/D)、髓过氧化物酶(myeloperoxidase, MPO)、检测小鼠血清与MLE-12细胞上清液TNF-α、IL-1β和IL-6、超氧化物歧化酶(superoxide dismutase, SOD)、丙二醛(malondialdehyde, MDA),H-E染色观察小鼠肺组织病理改变，Western Blot检测小鼠与MLE-12细胞NOD样受体家族含pyrin结构域蛋白3(NOD-like receptor family pyrin domain-containing protein 3, NLRP3)/凋亡相关颗粒样蛋白(apoptosis-associated speck-like protein containing a CARD, ASC)/Caspase-1/IL-1β通路蛋白表达。构建NLRP3沉默的MLE-12细胞株验证SAB的作用。结果：与空白组比较，LPS组小鼠体质量在造模后显著下降，而LPS+SAB组差异无统计学意义。与空白组比较，LPS组小鼠WBC、NEU、LYM水平均升高。与LPS组比较，LPS+SAB组WBC、NEU、LYM显著降低。与空白组比较，LPS组肺功能指标RL、Re水平升高，Cydn、PEF、MVV水平降低，与LPS组比较，LPS+SAB组RL、Re水平显著降低，Cydn、PEF、MVV显著升高。SAB显著降低脓毒症小鼠肺W/D和MPO。SAB显著降低了小鼠血清和MLE-12细胞上清液中TNF-α、IL-1β和IL-6水平、升高SOD水平与降低MDA水平。Western Blot实验显示SAB显著降低小鼠肺组织和MLE-12细胞中的NLRP3、ASC、Caspase-1和IL-1β的蛋白表达水平。当沉默NLRP3后，SAB的药效显著增强。结论：SAB显著减轻LPS诱导的ALI，其机制与抑制NLRP3相关，为治疗脓毒症ALI提供了潜在的药物。

Objective: To investigate the effect of salvianolic acid B(SAB) on lipopolysaccharide(LPS) induced acute lung injury(ALI). Methods: In vivo, a LPS-induced mouse model was used, divided into five groups: blank group, LPS group(10 mg/kg), LPS+dexamethasone(Dex) group[LPS+Dex(2 mg/kg)], LPS+SAB low-dose group(25 mg/kg), and LPS+SAB high-dose group(50 mg/kg).In vitro, LPS-induced mouse lung epithelial cell MLE-12 cell model was used, divided into blank group, LPS group(50 mmol/L),LPS+Dex group(1 μmol/L), LPS+SAB low-dose group(10 μmol/L), and LPS+SAB high-dose group(20 μmol/L). 24 hours after the last administration, blood routine indicators[including total WBC, neutrophils(NEU), lymphocytes(LYM)], lung function indicators[including lung resistance(RL), pulmonary expiratory resistance(Re), lung dynamic compliance(Cydn), peak expiratory flow rate(PEF), and maximum ventilation volume(MVV)] were measured in each group of mice. The dry wet ratio(W/D) of mouse lung, myeloperoxidase(MPO), TNF-α, IL-1β, IL-6 and superoxide dismutase(SOD), malondialdehyde(MDA), in the supernatant of mouse and MLE-12 cells were detected. H-E staining was used to detect the pathological changes of lung tissue in mice and Western Blot were used to detect the expression of NOD-like receptor family pyrin domain-containing protein 3(NLRP3)/apoptosis-associated speck-like protein containing a CARD(ASC)/Caspase-1/IL-1β pathway protein in mice and MLE-12 cells. Construct an NLRP3 silenced MLE-12 cell line to verify the role of SAB. Results: Compared with the blank group,the weight of the LPS group mice significantly decreased during modeling. There was no significant difference in the LPS+SAB group. Compared with the blank group, the levels of WBC, NEU, and LYM in the LPS group mice were increased. Compared with the LPS group, LPS+SAB significantly reduced WBC, NEU, and LYM. Compared with the blank group, the lung function indicators RL and Re levels in the LPS group increased, while Cydn, PEF, and MVV levels decreased. Compared with the LPS group, LPS+SAB reduced RL and Re levels, and increased Cydn, PEF, and MVV significantly. SAB significantly reduced lung W/D and MPO in septic mice. SAB significantly reduced TNF-α, IL-1β and IL-6 levels in mouse serum and MLE-12 cell supernatant, increased SOD levels, and decreased MDA levels. Western Blot experiments showed that SAB significantly reduced the protein expression of NLRP3, ASC, Caspase-1 and IL-1β in mouse lung tissue and MLE-12 cells. Furthermore,it was found that when NLRP3 was silenced, the efficacy of SAB was significantly enhanced. Conclusion: SAB significantly alleviates LPS induced ALI, and its mechanism is related to the inhibition of NLRP3, providing potential drugs for the treatment of sepsis induced ALI.