Research on the material basis of anti inflammation of Zhimu Huangbai medicine based on network pharmacology, molecular docking, and experimental verification
Inflammation is a complex interaction between soluble factors and cells that occurs in the body after trauma, infection, or autoimmune damage. The occurrence of inflammation is related to multiple factors, and the invasion of pathogens such as viruses, bacteria, and fungi can induce inflammation in the body. Once inflammation occurs, it may trigger serious medical consequences. Studies have shown that chronic or frequent inflammation can trigger cancer and autoimmune diseases.
A medicinal pair is a compatibility unit between a single herb and a compound, which has the characteristics of flexible addition and subtraction in compound medicine, as well as the relatively simple chemical composition of a single herb. Zhimu Huangbai Medicine is from the “Secret Collection of the Orchid Chamber” and has the effects of nourishing yin, clearing heat, purging fire, and detoxifying. It is mainly used for treating Yin deficiency and excessive fire. Anemarrhena Rhizoma Phellodendri Phellodendri pair is a common pair of drugs in clinical practice, and many prescriptions and traditional Chinese patent medicines and simple preparations contain this compatibility. Research shows that this drug has certain anti-inflammatory effect on or including the prescriptions of this drug pair. Mao et al. found that Zhibai Dihuang Pill can regulate inflammatory factors and promote the healing of inflammatory response injuries. Zuo et al. found through clinical research that the Sanhua decoction containing Zhimu Huangbai was effective in treating 108 cases of chronic prostatitis, with an effective rate of 83.5%. And modern pharmacological studies have shown that Zhimu Huangbai medicine also has significant therapeutic effects on chronic prostatitis. Our preliminary research has shown that Zhimu Huangbai medicine has a significant anti-inflammatory effect on the 30% ethanol elution site of the water extract at the cellular level. In order to further explore the anti-inflammatory mechanism of Zhimu Huangbai, this study intends to first predict the anti-inflammatory mechanism of Zhimu Huangbai through network pharmacology and molecular docking technology, and verify it through a cellular inflammation model. This study will provide some reference for the clinical application and basic research of Zhimu Huangbai medicine.

Our research group found in the early stage that Zhimu Huangbai has significant anti-inflammatory effects. In order to explore its material basis and possible mechanism of anti-inflammatory effects, this study first screened the core chemical components of Zhimu Huangbai’s anti-inflammatory effects, including quercetin, β – sitosterol, and dehydrated gibberellin, through network pharmacology. Studies have shown that quercetin can inhibit inflammation in mouse RAW264.7 cells through the TLR4/NF – κ B signaling pathway, and effectively alleviate inflammatory factors in human rheumatoid arthritis fibroblast like synovial cells by inhibiting the expression of NF – κ B. β – sitosterol has good anti-inflammatory activity against yeast induced fever in rats in beer. Dehydrated icariin has a significant protective effect on yeast polysaccharide induced peritonitis in mice, which may be related to its inhibition of macrophage calcium influx and iNOS expression. The molecular docking results showed that dehydrated icariin has high binding activity with PPARG, β – sitosterol, and quercetin, all of which bind to NR3C1. Among them, dehydrated icariin scored the highest in the docking results with PPARG. This suggests that dehydrated icariin may be the main active ingredient in the anti-inflammatory properties of Anemarrhena henryi and Phellodendron amurense.
Based on the prediction results of network pharmacology and molecular docking, this study detected the content of dehydrated icariin in Zhimu Huangbai. The results showed that the content of dehydrated icariin in Zhimu Huangbai was 18.04%. To further investigate the anti-inflammatory activity and possible mechanisms of Zhimu Huangbai and dehydrated icariin, we studied the effects of Zhimu Huangbai and dehydrated icariin on the production of related inflammatory factors at the cellular level. The results showed that Zhimu Huangbai and dehydrated icariin could significantly inhibit LPS induced TNF – α, IL-6, and NO production. The TNF signaling pathway is the most important pathway predicted by KEGG for the anti-inflammatory effect of Zhimu Huangbai. Tumor necrosis factor alpha (TNF – α) is one of the tumor necrosis factor receptors (TNFR), and the peroxisome proliferator activated receptor (PPAR) is a key transcriptional regulator in the nuclear receptor superfamily. Research has shown that TNF – α can inhibit the activity of peroxisome proliferator activated receptor gamma (PPAR gamma), reduce the expression of PPAR gamma, and further activate the NF – κ B pathway in a PPAR gamma dependent manner. NF – κ B further promotes the expression of downstream inflammation related genes such as IL-6 and iNOS, while NO is catalyzed by iNOS. Therefore, this study speculates that the anti-inflammatory effect of Zhimu Huangbai through the TNF pathway is one of its important mechanisms.
This study screened the anti-inflammatory components, targets, and signaling pathways of Zhimu Huangbai through network pharmacology. Molecular docking technology was used to verify the binding ability of dehydrated icariin to core targets, and the content of dehydrated icariin in Zhimu Huangbai was determined through content determination. Cell experiments were conducted to verify the anti-inflammatory effects of Zhimu Huangbai and dehydrated icariin, and their anti-inflammatory mechanisms were preliminarily explored. This study provides a reference method for the material basis and mechanism research of drug intervention in diseases, and provides a research basis for the clinical use of Zhimu Huangbai.