Innovation Research Institute of Traditional Chinese Medicine (IRI), Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
Integrated Traditional Chinese and Western Medicine Hospital, Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
Comb Chem High Throughput Screen. 2023;26(2):392-409. doi: 10.2174/1386207325666220418095316.
The "Ruan Jian Qing Mai (RJQM) recipe" is a traditional Chinese medicine (TCM), which has been found to have significant curative effects on diabetic ulcers in the clinic for a long time. Previous research has shown that RJQM can improve diabetic skin wound healing and promote angiogenesis. However, the active ingredients of the RJQM recipe and its pharmacological mechanism of treatment for diabetic skin wound healing still remain unclear.This study aims to investigate the effect of the RJQM recipe on diabetic wound healing, and to identify the possible active ingredients and their mechanism.
First, a skin injury model was established in diabetic mice, and wound healing was evaluated by hematoxylin-eosin (HE) staining, quantitative reverse transcription-polymerase chain reaction (RT-qPCR), and western blot analysis. Second, the chemical constituents of the RJQM recipe were analyzed and identified by ultra pressure liquid chromatography-mass spectrometry (UPLC-MS). Finally, the possible targets of drug treatment for diabetic skin injury were analyzed by network pharmacology and verified by in vitro experiments using cell culture.
(1) In the full-thickness skin injury model, the skin wound healing rate and healing area were significantly increased in mice treated with the RJQM recipe compared with those of the model group. The results of immunofluorescence staining showed that the RJQM recipe could increase the expression of VEGF protein and promote the proliferation of vascular smooth muscle cells and the formation of microvessels, and RT-qPCR results found that the mRNA expression of angiogenesis-related factors in the RJQM recipe group was significantly higher than that in the model group. (2) A total of 25 compounds were identified by UPLC-MS. (3) According to the results of network pharmacology, the therapeutic effect of the RJQM recipe on diabetic skin injury may be related to S6 (quercetin), S1 (typhaneoside), S18 (isoliquiritigenin), protein kinase B-α (Akt1), phosphoinositide-3-kinase regulatory subunit 1 (PIK3R1), insulin-like growth factor I receptor (IGF1R), vascular endothelial growth factor-a (VEGF-a), signal transducer and activator of transcription-3 (STAT3) and phosphoinositide 3-kinase-protein kinase B (PI3K-Akt) signaling pathways. Based on the predictions by network pharmacology, we proved that the drug could treat diabetic skin damage by activating the PI3K-Akt-VEGF signaling pathway.
The RJQM recipe promotes the formation of granulation tissue during the process of wound healing and exerts a good therapeutic effect on diabetic skin wound healing.
“软煎青盲（RJQM）方”是一种中药（TCM），在临床上发现对糖尿病溃疡有显著的疗效。先前的研究表明，RJQM 可以改善糖尿病皮肤伤口愈合，促进血管生成。然而，RJQM 方的活性成分及其治疗糖尿病皮肤伤口愈合的药理机制仍不清楚。本研究旨在探讨 RJQM 方对糖尿病伤口愈合的影响，并确定可能的活性成分及其机制。
首先，在糖尿病小鼠中建立皮肤损伤模型，通过苏木精-伊红（HE）染色、定量逆转录-聚合酶链反应（RT-qPCR）和 Western blot 分析评估伤口愈合情况。其次，采用超高效液相色谱-质谱联用（UPLC-MS）分析 RJQM 方的化学成分，并进行鉴定。最后，通过网络药理学分析糖尿病皮肤损伤药物治疗的可能靶点，并通过细胞培养的体外实验进行验证。
（1）在全层皮肤损伤模型中，与模型组相比，RJQM 方治疗的小鼠皮肤伤口愈合率和愈合面积显著增加。免疫荧光染色结果表明，RJQM 方可以增加 VEGF 蛋白的表达，促进血管平滑肌细胞的增殖和微血管的形成，RT-qPCR 结果发现 RJQM 方组血管生成相关因子的 mRNA 表达明显高于模型组。（2）通过 UPLC-MS 共鉴定出 25 种化合物。（3）根据网络药理学的结果，RJQM 方治疗糖尿病皮肤损伤的疗效可能与 S6（槲皮素）、S1（知母皂苷 BⅡ）、S18（异甘草素）、蛋白激酶 B-α（Akt1）、磷酸肌醇-3-激酶调节亚基 1（PIK3R1）、胰岛素样生长因子 I 受体（IGF1R）、血管内皮生长因子-a（VEGF-a）、信号转导和转录激活因子 3（STAT3）和磷酸肌醇 3-激酶-蛋白激酶 B（PI3K-Akt）信号通路有关。基于网络药理学的预测，我们证明了该药物可以通过激活 PI3K-Akt-VEGF 信号通路来治疗糖尿病皮肤损伤。
RJQM 方促进肉芽组织形成，对糖尿病皮肤伤口愈合有较好的治疗效果。
