He Xu, Chen Guo-Feng, Tao Wei-Ting, Huang Xiao-Jia, Lin Ye, Sun Jing, Li Yan
School of Pharmacy & School of Biological and Food Engineering, Changzhou University, Changzhou, China.
School of Medical and Health Engineering, Changzhou University, Changzhou, China.
J Thorac Dis. 2025 Mar 31;17(3):1605-1616. doi: 10.21037/jtd-24-1561. Epub 2025 Mar 26.
Tetramethylpyrazine (TMP) possesses anti-inflammatory and antioxidant properties and plays a crucial role in mitigating acute lung injury (ALI). However, the specific underlying mechanisms remain elusive. High mobility group box 1 (HMGB1), a pro-inflammatory factor, can bind to Toll-like receptor 4 (TLR4), activating downstream signaling pathways, which in turn activate nuclear factor kappa-B (NF-κB). This study aims to explore the preventive effects of TMP on lipopolysaccharide (LPS)-induced ALI and its influence on the HMGB1/TLR4/NF-κB signaling pathway.
C57BL/6 mice were randomly divided into a control group (CON group), a model group (LPS group), a heparin (Hep) group, and a TMP group. In the model group, mice received an initial intraperitoneal injection of LPS followed by a second airway injection three hours later to induce ALI. In the TMP group, LPS was administered 30 minutes before the first intraperitoneal injection and followed by intratracheal atomization of LPS. TMP was injected intraperitoneally 30 minutes after the second LPS administration. The Hep group received heparin at 200 IU/kg following the same schedule as the TMP group. Peripheral capillary oxygen saturation (SpO) was measured 24 hours after the initial intraperitoneal injection. Lung tissues were harvested for wet to dry weight ratio (W/D) calculation and pathological assessment via hematoxylin and eosin (HE) staining. Western blot analysis was performed to evaluate the protein levels of HMGB1, TLR4, and NF-κB p65 in lung tissues, along with the assessment of leukocyte content and protein concentration in bronchoalveolar lavage fluid (BALF).
Compared to the control group, the model group showed reduced SpO levels, increased inflammatory indicators, and elevated expression levels of HMGB1, TLR4, and phosphorylated p65 (p-p65) proteins. TMP treatment led to a decrease in the W/D, reduced alveolar exudation, improved alveolar structure, and decreased total white blood cell count and protein concentration in BALF. Notably, the protein levels of HMGB1, TLR4, and p-p65 were significantly reduced in the TMP and Hep groups, while there were no significant differences in p65 expression among the groups.
TMP can alleviate LPS-induced ALI in mice by reducing lung inflammatory response through inhibiting the HMGB1/TLR4/NF-κB pathway.
川芎嗪（TMP）具有抗炎和抗氧化特性，在减轻急性肺损伤（ALI）中起关键作用。然而，具体的潜在机制仍不清楚。高迁移率族蛋白B1（HMGB1）作为一种促炎因子，可与Toll样受体4（TLR4）结合，激活下游信号通路，进而激活核因子κB（NF-κB）。本研究旨在探讨TMP对脂多糖（LPS）诱导的ALI的预防作用及其对HMGB1/TLR4/NF-κB信号通路的影响。
将C57BL/6小鼠随机分为对照组（CON组）、模型组（LPS组）、肝素（Hep）组和TMP组。模型组小鼠先腹腔注射LPS，3小时后再气道注射一次以诱导ALI。TMP组在首次腹腔注射前30分钟给予LPS，随后气管内雾化LPS。在第二次给予LPS后30分钟腹腔注射TMP。Hep组按照与TMP组相同的给药方案给予200 IU/kg肝素。在首次腹腔注射后24小时测量外周毛细血管血氧饱和度（SpO）。采集肺组织计算湿干重比（W/D），并通过苏木精-伊红（HE）染色进行病理评估。进行蛋白质印迹分析以评估肺组织中HMGB1、TLR4和NF-κB p65的蛋白水平，同时评估支气管肺泡灌洗液（BALF）中的白细胞含量和蛋白质浓度。
与对照组相比，模型组SpO水平降低，炎症指标升高，HMGB1、TLR4和磷酸化p65（p-p65）蛋白表达水平升高。TMP治疗导致W/D降低，肺泡渗出减少，肺泡结构改善，BALF中白细胞总数和蛋白质浓度降低。值得注意的是，TMP组和Hep组中HMGB1、TLR4和p-p65的蛋白水平显著降低，而各组之间p65表达无显著差异。
TMP可通过抑制HMGB1/TLR4/NF-κB通路减轻肺部炎症反应，从而缓解LPS诱导的小鼠ALI。
