Elucidation of Oxymatrine’s Therapeutic Mechanism in Cryptosporidiosis via TNF/NF-kappaB Signaling Pathway: An Integrated Approach Utilizing Network Pharmacology and Experimental Validation

About the Study

A study published in scientific reports in June 2024 investigates the mechanism of oxymatrine in treating cryptosporidiosis via the TNF/NF-kappaB signaling pathway. The research team utilized network pharmacology and animal experiments to identify the key targets and signaling pathways involved in oxymatrine’s therapeutic action against cryptosporidiosis. The researchers employed a mouse model of cryptosporidiosis to validate the involvement of the TNF/NF-κB signaling pathway, elucidating the efficacy and potential mechanism of oxymatrine in treating this parasitic infection. This comprehensive approach, combining network pharmacology with experimental validation, not only advances our understanding of oxymatrine's role in cryptosporidiosis treatment but also presents novel therapeutic strategies. The study presents a significant contribution to the field of traditional Chinese medicine modernization, bridging the gap between ancient remedies and contemporary scientific methodologies.

Cryptosporidiosis (CPS), a zoonotic parasitic disease caused by Cryptosporidium species, primarily manifests as acute diarrhea. The World Health Organization has classified CPS as one of the six major global diarrheal diseases, and it is a significant cause of mortality in AIDS patients [1]. Moreover, CPS ranks as the second leading cause of diarrhea in children under five years of age [2-4]. The potential for large-scale epidemics exists due to the contamination of water sources by Cryptosporidium oocysts, presenting a substantial public health concern [5]. Despite considerable progress in understanding the etiology, molecular biology and comprehensive prevention and control technologies for Cryptosporidium, the immunoregulatory mechanisms underlying infection remain elusive. Furthermore, there is a notable absence of effective therapeutic agents or preventive vaccines. Consequently, elucidating the molecular mechanism of Cryptosporidium parvum infection and developing efficacious therapeutic interventions with minimal adverse effects have emerged as critical priorities in the scientific community.

Over an extended period, researchers have investigated more than 200 compounds for the treatment of cryptosporidiosis, including anticoccidial agents, antiprotozoal drugs, anthelminthics, broad-spectrum antibiotics, biological agents and parasite protein kinase inhibitors. However, none of these interventions have demonstrated effective control of Cryptosporidium infection [6]. Nitazoxanide, approved by the U.S. Food and Drug Administration for treating cryptosporidiosis in young children, remains the sole authorized therapeutic option. Nevertheless, its efficacy is limited in immunocompromised or immunodeficient patients, such as those with AIDS, organ transplant recipients and malnourished children [7]. This underscores the ongoing challenge in developing effective treatments for cryptosporidiosis, particularly for vulnerable populations.

Recent research has highlighted the potential of Traditional Chinese Medicine (TCM) in addressing cryptosporidiosis, owing to its diverse resources, multi-target mechanisms of action and relatively low incidence of adverse effects. Studies have demonstrated the efficacy of various TCM compounds, such as allicin, Sophora flavescens and Astragalus, in treating cryptosporidiosis [6]. Sophora flavescens Ait., a member of the Fabaceae family, has been utilized in TCM since its first mention in the "Shennong's Classic of Materia Medica" during the Han Dynasty. The dried roots of this plant, known as Sophora flavescens in TCM, are characterized by their cold nature and bitter taste. They are traditionally employed for their heat-clearing, dampness-drying, anti-inflammation and antiparasitic properties. Previous investigations by our research team have revealed that Sophora flavescens mixture exhibits promising therapeutic effects against cryptosporidiosis. Its mechanism of action primarily involves inhibition and elimination of Cryptosporidium parvum, coupled with enhancement of intestinal mucosal immune function. Oxymatrine (OMT), the principal bioactive monomer in Sophora flavescens, has garnered significant attention due to its diverse pharmacological effects, including anti-tumor, anti-fibrotic, antiviral and anti-inflammatory properties [8]. The relative safety, cost-effectiveness and accessibility of OMT have contributed to its emergence as a focal point in TCM monomer compound research. Our team’s findings indicate that OMT can facilitate the repair of Cryptosporidium-infected intestinal mucosa through modulation of Toll-Like Receptors (TLRs) in murine intestinal mucosal tissues [9]. Given these promising results, elucidating the precise mechanisms underlying OMT’s therapeutic action in cryptosporidiosis has become a critical research objective.

This study employed network pharmacology and molecular docking technologies to investigate the multi-target and multi-pathway synergistic anti-infection mechanisms of TCM in cryptosporidiosis treatment. Initially, bioinformatics techniques and data resources were utilized to predict the core targets (RELA, AKT1, TNF, CASP3, IL-6) and principal signaling pathways involved in OMT’s therapeutic action against cryptosporidiosis. The research focused on the role of the TNF/NF-κB signaling pathway in cryptosporidiosis treatment. TNF and NF-κB are crucial mediators of inflammatory responses and immune regulation, and network pharmacology analysis suggested their significant involvement in Cryptosporidium infection processes. Experimental validation, a critical component of the study, involved the development of a murine cryptosporidiosis model to examine OMT’s effects on the TNF/NF-κB signaling pathway and elucidate its therapeutic mechanism. Results demonstrated that OMT inhibits the TNF/NF-κB signaling pathway and reduces expression levels of related factors TNF-α, NF-κB, and IL-6, thereby modulating intestinal inflammation and effectively treating cryptosporidiosis. In conclusion, OMT, as a TCM component with diverse biological activities, demonstrates significant potential in the treatment of cryptosporidiosis. However, current research remains in its nascent stages, necessitating more in-depth and comprehensive investigations. Future studies should focus on elucidating the specific mechanisms by which OMT modulates the TNF/NF-κB pathway to affect Cryptosporidium infection processes, thereby more precisely defining its potential targets and mode of action in cryptosporidiosis treatment. It is important to note that in the anti-infection process involves complex interactions and overlaps among various signaling pathways, resulting in synergistic signal transmission effects. This complexity warrants further exploration to fully understand the multi-faceted action of OMT. Given that cryptosporidiosis represents a global public health concern, future research efforts should prioritize international collaboration and knowledge exchange. Such cooperation can leverage the unique advantages of TCM in providing characteristic treatments, potentially leading to the development of novel strategies and pharmaceutical interventions for the effective prevention and management of cryptosporidiosis on a global scale.

Competing Interests

The authors declare no competing interests.