本研究旨在探讨黄连碱在急性肺损伤（ALI）中的作用及机制。采用脂多糖（LPS）刺激小鼠肺上皮12（MLE-12）细胞，构建体外肺损伤模型，研究黄连碱的作用。脓毒症引起的 ALI。通过细胞计数试剂盒 8 测定评估 MLE-12 细胞的活力。通过酶联免疫吸附血清学测定测定肿瘤坏死因子-α (TNF-α)、白细胞介素 6 (IL-6) 和 IL-1β 的细胞因子释放。通过逆转录定量聚合酶链反应检测TNF-α、IL-6和IL-1β mRNA的相对表达水平。通过Annexin V/碘化丙啶染色测定MLE-12细胞的细胞凋亡，并通过流式细胞仪进行分析。 Western blot分析观察凋亡相关蛋白Bax和Cleaved Caspase-3的表达。通过Western blot检测B细胞抑制剂α（IκBα）中的磷酸化p65、p65、磷酸化核因子κ轻链多肽基因增强子（IκBα）和IκBα，发现核因子κB（NF-κB）信号通路被激活。印迹分析。黄连碱处理可以显着恢复LPS刺激引起的MLE-12细胞活力下降。黄连碱治疗显着抑制TNF-α、IL-6和IL-1β的释放。黄连碱处理可抑制LPS诱导的MLE-12细胞凋亡，并抑制Bax和cleaved Caspase-3的表达水平。黄连碱联合LPS刺激，显着降低了磷酸-IκBα的蛋白水平，增加了IκBα的水平，并降低了磷酸-p65-p65的比率。这些结果表明，黄连碱通过抑制肺上皮细胞炎症和凋亡来减轻脓毒症肺损伤。 NF-κB 通路。因此，黄连碱可能具有治疗脓毒症引起的 ALI 的潜力。

This study aimed to investigate the functioning and mechanism of coptisine in acute lung injury (ALI).Murine Lung Epithelial 12 (MLE-12) cells were stimulated with lipopolysaccharide (LPS) to construct an in vitro pulmonary injury model to study the functioning of coptisine in sepsis-induced ALI. The viability of MLE-12 cells was assessed by the cell counting kit-8 assay. The cytokine release of tumor necrosis factor-α (TNF-α), interleukin 6 (IL-6), and IL-1β was measured by enzyme-linked-immunosorbent serologic assay. The relative expression levels of TNF-α, IL-6, and IL-1β mRNA were examined by reverse transcription-quantitative polymerase chain reaction. The cell apoptosis of MLE-12 cells was determined by Annexin V/propidium iodide staining and analyzed by flow cytometry. The expressions of apoptosis-related proteins Bax and cleaved Caspase-3 were observed by Western blot analysis. The activation of nuclear factor kappa B (NF-κB) signaling pathway was discovered by the determination of phospho-p65, p65, phospho-nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor alpha (IκBα), and IκBα through Western blot analysis.Coptisine treatment could significantly restore decrease in MLE-12 cell viability caused by LPS stimulation. The release of TNF-α, IL-6, and IL-1β was significantly inhibited by coptisine treatment. Coptisine treatment inhibited MLE-12 cell apoptosis induced by LPS, and also inhibited the expression levels of Bax and cleaved Caspase-3. Coptisine treatment along with LPS stimulation, significantly reduced the protein level of phospho-IκBα, increased the level of IκBα, and reduced phospho-p65-p65 ratio.These results indicated that coptisine attenuated sepsis lung injury by suppressing lung epithelial cell inflammation and apoptosis through NF-κB pathway. Therefore, coptisine may have potential to treat sepsis-induced ALI.