Department of Respiratory and Critical Care Medicine, Renmin Hospital of Wuhan University, Wuhan, China.
Department of Emergency, Renmin Hospital of Wuhan University, Wuhan, China.
J Pharm Pharmacol. 2022 Aug 19;74(8):1117-1124. doi: 10.1093/jpp/rgac035.
Our previous study found that Lianhuaqingwen reduces lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice by suppressing p53-mediated apoptosis. To identify the type of lung cells affected by Lianhuaqingwen, we conducted a cell experiment.
C57/B6 mice and A549 cells were administered Lianhuaqingwen and LPS. A549 cells were transfected with p53 siRNA to inhibit p53. The degree of ALI in mice was validated by haematoxylin and eosin staining as well as measurement of IL-1β and MCP-1 levels. In A549 cells, Cell Counting Kit-8 (CCK-8), DHE and TUNEL assays were used to assess cell viability, reactive oxygen species (ROS) production and apoptosis, respectively. Western blot analysis was used to evaluate the protein expression of p53, Bcl-2, Bax, caspase-9 and caspase-3. Co-immunofluorescence was used to detect cytochrome C distribution.
Lianhuaqingwen alleviated LPS-induced ALI in vivo. Lianhuaqingwen at 300 μg/ml increased cell viability, lowered ROS production and reduced apoptotic cells in vitro. Lianhuaqingwen enhanced Bcl-2 expression and reduced Bax, caspase-9 and caspase-3 expression as well as blocked cytochrome C release under LPS stimulation. Treatment with a combination of Lianhuaqingwen and p53 siRNA was more effective than treatment with Lianhuaqingwen alone.
Lianhuaqingwen inhibits p53-mediated apoptosis in alveolar epithelial cells, thereby preventing LPS-induced ALI.
我们之前的研究发现，连花清瘟通过抑制 p53 介导的细胞凋亡来减轻脂多糖（LPS）诱导的急性肺损伤（ALI）。为了确定连花清瘟作用的肺细胞类型，我们进行了细胞实验。
给 C57/B6 小鼠和 A549 细胞施用连花清瘟和 LPS。用 p53 siRNA 转染 A549 细胞以抑制 p53。通过苏木精和伊红染色以及测量白细胞介素-1β（IL-1β）和单核细胞趋化蛋白-1（MCP-1）水平来验证小鼠的 ALI 程度。在 A549 细胞中，使用细胞计数试剂盒-8（CCK-8）、二氢乙锭（DHE）和 TUNEL 测定法分别评估细胞活力、活性氧（ROS）产生和细胞凋亡。使用 Western blot 分析评估 p53、Bcl-2、Bax、caspase-9 和 caspase-3 的蛋白表达。共免疫荧光检测细胞色素 C 分布。
连花清瘟在体内减轻 LPS 诱导的 ALI。连花清瘟在 300μg/ml 时增加细胞活力，降低 ROS 产生并减少体外 LPS 刺激下的凋亡细胞。连花清瘟增强 Bcl-2 的表达，降低 Bax、caspase-9 和 caspase-3 的表达，并阻断细胞色素 C 的释放。与单独使用连花清瘟相比，联合使用连花清瘟和 p53 siRNA 的治疗效果更显著。
连花清瘟抑制肺泡上皮细胞中 p53 介导的细胞凋亡，从而预防 LPS 诱导的 ALI。
