Yang Lan, Wang Xiu-Wen, Zhou Bing, Yang En-Hua, Gong Wen-Qiao, Yao Jing-Chun, Sun Cheng-Hong, Pan Li-Hong, Cheng Guo-Liang
School of Pharmacy, Shandong University of Traditional Chinese Medicine Ji'nan 250355, China.
State Key Laboratory of Integration and Innovation of Classic Formula and Modern Chinese Medicine,Lunan Pharmaceutical Group Co., Ltd. Linyi 276005, China.
Zhongguo Zhong Yao Za Zhi. 2024 Jun;49(12):3302-3311. doi: 10.19540/j.cnki.cjcmm.20240208.401.
This study aims to investigate the mechanism of Mailuo Shutong Pills(MLST) on posterior limb muscle swelling caused by femoral fracture(SCFF) through network pharmacology and animal experiments. The plasma components of MLST were analyzed by LC-MS, and the target and signal pathway of SCFF were predicted by network pharmacology and verified by molecular docking. SCFF model rats were established through animal experiments, and different doses of MLST were administered to detect the degree of limb swelling. Hematoxylin-eosin(HE) staining was used to observe pathological changes in muscle tissue, and interleukin-6(IL-6), interleukin-1β(interleukin-1β), and tumor necrosis factor-α(TNF-α) in peripheral blood were determined by enzyme-linked immunosorbent assay(ELISA). The expression of relevant signaling pathways was measured by Western blot. Network pharmacological results showed that MLST and SCFF had a total of 153 disease targets, and the key targets were IL-6, TNF, etc., involving mitogen-activated protein kinase(MAPK) signaling pathway, phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT) signaling pathway, etc. The binding energies of the main components and key targets were lower than-7.0 kcal·mol~(-1), indicating that the network analysis results were reliable. The results of animal experiments showed that MLST could reduce the swelling degree and pathological damage of the posterior limb muscles of SCFF rats compared with the model group. ELISA results showed that MLST could reduce the levels of IL-6, IL-1β, and TNF-α in the serum of SCFF rats. Western blot results showed that MLST can reduce the expression of p-AKT, p-PI3K, p-NF-κB, p-p38 MAPK, and p-ERK in SCFF rats. MLST may reduce the content of inflammatory factors in serum by regulating the expression of PI3K/AKT and MAPK-related signaling pathway protein and improving posterior limb muscle SCFF in rats.
本研究旨在通过网络药理学和动物实验探讨脉络舒通丸（MLST）对股骨骨折致后肢肌肉肿胀（SCFF）的作用机制。采用液相色谱-质谱联用（LC-MS）分析MLST的血浆成分，通过网络药理学预测SCFF的靶点和信号通路，并进行分子对接验证。通过动物实验建立SCFF模型大鼠，给予不同剂量的MLST，检测肢体肿胀程度。采用苏木精-伊红（HE）染色观察肌肉组织病理变化，采用酶联免疫吸附测定（ELISA）法检测外周血白细胞介素-6（IL-6）、白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）水平。采用蛋白质免疫印迹法检测相关信号通路的表达。网络药理学结果显示，MLST与SCFF共有153个疾病靶点，关键靶点为IL-6、TNF等，涉及丝裂原活化蛋白激酶（MAPK）信号通路、磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（AKT）信号通路等。主要成分与关键靶点的结合能均低于-7.0 kcal·mol~(-1)，表明网络分析结果可靠。动物实验结果显示，与模型组相比，MLST可减轻SCFF大鼠后肢肌肉的肿胀程度和病理损伤。ELISA结果显示，MLST可降低SCFF大鼠血清中IL-6、IL-1β和TNF-α水平。蛋白质免疫印迹结果显示，MLST可降低SCFF大鼠中p-AKT、p-PI3K、p-NF-κB、p-p38 MAPK和p-ERK的表达。MLST可能通过调节PI3K/AKT和MAPK相关信号通路蛋白的表达，降低大鼠血清中炎症因子含量，改善后肢肌肉SCFF。
