Non-clinical pharmacology of traditional Chinese medicine (TCM) formulae plays a crucial role in elucidating their therapeutic mechanisms. With the integration of cutting-edge disciplines such as systems biology, big data, and artificial intelligence, the research depth and breadth in this field continue to expand. However, technical standards for non-clinical pharmacology research on TCM formulae have not yet been established. To address this, technical guidelines for non-clinical pharmacology research of TCM formulae were formed, aiming to scientifically standardize research content, elevate research quality, and facilitate the mutual translation between clinical and basic research. Guided by integrative pharmacology of TCM and driven by clinical value, these guidelines employ multi-dimensional qualitative pharmacokinetics (PK) and pharmacodynamics (PD) studies alongside quantitative "PK-PD" studies of key components. This approach enables the transition from panoramic description to precise mechanistic research, establishing a complete evidence chain system for the effectiveness of TCM formulae and demonstrating their holistic action characteristics of "multi-component, multi-pathway, multi-target". Meanwhile, the guidelines emphasize that animal models for TCM research should ensure stability and reproducibility. Additionally, pharmacodynamic evaluation is characterized by its measurability and quantifiability. The mechanisms of action at the molecular level should be in-depth and interpretable. These aspects collectively provide guidance for basic non-clinical pharmacology research on TCM formulae and the development of new TCM drugs. The formulation of these guidelines leveraged the research strengths of the drafting units' teams, incorporated literature review findings, and achieved consensus through extensive discussions among multidisciplinary experts. Broad expert opinions from universities, research institutes, and enterprises engaged in new drug development were solicited, ensuring the guidelines' practicality, standardization, scientific rigor, and feasibility.

Shengjiang Xiexin Decoction (SXD) is a widely recognized formula in Traditional Chinese Medicine (TCM) for treating diarrhea and is commonly used in clinical practice. Clostridium difficile infection (CDI) is a type of antibiotic-associated diarrhea with a rising incidence rate that has severe consequences for humans. Recent clinical applications have found significant efficacy in using SXD as an adjunct to CDI treatment. However, the pharmacodynamic substance basis and therapeutic mechanism of SXD remain unclear. This study aimed to systematically analyze the metabolic mechanisms and key pharmacodynamic components of SXD in CDI mice by combining non-targeted metabolomics of Chinese medicine and serum medicinal chemistry. We established a CDI mouse model to observe the therapeutic effect of SXD on CDI. We investigated the mechanism of action and active substance composition of SXD against CDI by analyzing 16S rDNA gut microbiota, untargeted serum metabolomics, and serum pharmacochemistry. We also constructed a multi-scale, multifactorial network for overall visualization and analysis. Our results showed that SXD significantly reduced fecal toxin levels and attenuated colonic injury in CDI model mice. Additionally, SXD partially restored CDI-induced gut microbiota composition. Non-targeted serum metabolomics studies showed that SXD not only regulated Taurine and hypotaurine metabolism but also metabolic energy and amino acid pathways such as Ascorbate and aldarate metabolism, Glycerolipid metabolism, Pentose and glucuronate interconversions, as well as body and other metabolite production in the host. Through the implementation of network analysis methodologies, we have discerned that Panaxadiol, Methoxylutcolin, Ginsenoside-Rf, Suffruticoside A, and 10 other components serve as critical potential pharmacodynamic substance bases of SXD for CDI. This study reveals the metabolic mechanism and active substance components of SXD for the treatment of CDI mice using phenotypic information, gut microbiome, herbal metabolomics, and serum pharmacochemistry. It provides a theoretical basis for SXD quality control studies.

Triptolide (TP) exerts a promising effect in the treatment of ulcerative colitis (UC). However, its toxicity seriously hinders its application in the clinic. Previous studies indicated that dendritic cells (DCs) are the main target through which TP exerts its immunoregulatory effect. Thus, we designed an approach to target DCs in vitro to avoid the direct exposure of organs to TP. Our results revealed that DCs pretreated with TP (DCTP) exerted satisfactory therapeutic effects in mice with colitis, resulting in improved colonic inflammation and alleviated local lesion damage. In addition, no obvious toxicity was observed. DCTP also reshaped the immune milieu by decreasing CD4⁺ T cell numbers and increasing regulatory T cell numbers in the spleen, mesenteric lymph nodes, peripheral blood and colon; these effects were further confirmed in vitro. Downregulation of CD80/86, ICAM-1, MHCI, TLR2/4, TNF-α, and IL-6 expression and upregulation of programmed cell death ligand 1 (PDL1) and IL-10 expression were observed, indicating that DCs were converted into tolerogenic DCs. In conclusion, DCTP can effectively reduce toxicity and alleviate colonic inflammation and local lesion damage in mice with colitis. The immune mechanism underlying the effects of DCTP included the conversion of DCs into tolerogenic DCs and the alteration of T cell differentiation to produce immunoinhibitory rather than immunostimulatory T cells.