Exploring the underlying mechanism of Huanglian Jiedu Decoction on sepsis-induced coagulopathy via network pharmacology and experiment validation

Yiming ShenBing ShaoYing-hao PeiSuzhen ZhangHaiyi DingQiuhua ChenLinfeng DaiXiaowen ZhouYuanyuan GuXing WangHua Jiang^*

• Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, China

Background: Sepsis-induced coagulopathy (SIC) is a critical complication associated with sepsis, characterized by disruptive coagulation. This study aims to examine the therapeutic effects of Huanglian Jiedu Decoction (HLJDD) on SIC and elucidate the underlying mechanisms using network pharmacology methodologies.Methods: The cecal ligation and puncture (CLP) method was used to establish a septic rat model. The rats were divided into three groups: the sham group, the CLP group, and the HLJDD group. Mortality rates, vital signs, histopathological testing of the lung and kidney, blood cell counts, and coagulation function were assessed to evaluate the severity of coagulation disorders and inflammatory injury across the groups. Network pharmacology was used to identify candidate targets, and the potential mechanism of HLJDD in alleviating SIC was elucidated through experimental verification.Results: The results showed that HLJDD significantly improved the survival rate of rats with CLP-induced sepsis, reduced inflammatory response and lung, kidney pathological damage, and alleviated coagulation dysfunction. This suggests that HLJDD has a good effect on improving SIC. Network pharmacology identified a total of 87 active compounds as the main components of HLJDD. At the same time, 21 potential targets were considered as candidate targets of HLJDD against SIC. GO and KEGG enrichment analysis showed that the candidate targets were significantly associated with lipid metabolism, atherosclerosis, the AGE-RAGE signaling pathway in diabetic complications, pathways in cancer, and tuberculosis. Estrogen receptor 1(ESR1), Interleukin-6, and C-X-C Motif Chemokine Ligand 8 were considered to play a key role in the candidate targets. Molecular docking suggested that the main components of HLJDD had good binding affinity with these genes. Furthermore, ESR1 expression was markedly downregulated in both lung tissue and kidney tissue in the CLP group. HLJDD restored the decreased ESR1 expression in the lung and kidney tissues of septic rats. Western blot analysis showed that HLJDD can regulate the levels of ESR1, IL-6, and CXCL8 proteins.Conclusion: Our data demonstrated that HLJDD treatment ameliorates coagulopathy in SIC conditions, including reductions in PT and APTT levels and increases the Fib contents. Network pharmacology analysis suggests that these protective effects of HLJDD are associated with the modulation of ESR1.