Cell necrosis and neuroinflammation play an important role in brain injury induced by ischemic stroke. Previous studies reported that Taohong Siwu decoction (THSWD)can reduce heart muscle cell necrosis and has anti-inflammatory properties. In this study, we investigated the effects of THSWD on cell necrosis and neuroinflammation in a rat model of middle cerebral artery occlusion (MCAO). Thirty-six male Sprague-Dawley (SD) rats were randomly divided into three groups with 12 rats in each group. They were the sham operation group, MCAO model group, and MCAO + THSWD group. We used ELISA to determine the levels of TNF-α, Mcp-1, and IL-1β inflammatory factors in rat serum, qRT‐PCR to detect the expression of TNF‐α, Mcp‐1 and IL‐1β mRNA in rat brain, and immunohistochemistry to detect the number of microglia and neutrophils in rat brain. qRT-PCR and Western blot were used to detect the mRNA and protein expression levels of IBA-1 and MPO inflammatory factors and the TNF-α/RIP1/RIP3/MLKL pathway in the rat brain and protein expression levels. Compared with the sham operation group, the expression of MCP-1, IL-1β, IBA-1, and MPO inflammatory factors and the TNF-α/RIP1/RIP3/MLKL pathway were significantly upregulated in the MCAO group. Compared with the MCAO group, the expressions of MCP-1, IL-1β, IBA-1, and MPO inflammatory factors and the TNF-α/RIP1/RIP3/MLKL pathway were significantly downregulated in the MCAO + THSWD group. THSWD can reduce the expression levels of MCP-1, IL-1β, IBA-1, and MPO inflammatory factors as well as the TNF-α/RIP1/RIP3/MLKL pathway. Meanwhile, it can reduce the necrosis and inflammation of brain cells after cerebral ischemia, so as to protect the brain tissue of rats.

Background and aims: Taohong Siwu Decoction (THSWD) is a traditional Chinese herbal prescription that is effective for ischemic stroke, Whether THSWD regulates the CCL2/CCR2 axis and thus reduces the inflammatory response induced by ischemic stroke is not known. The aim of this study was to elucidate the mechanism of action of THSWD in the treatment of ischemic stroke using bioinformatics combined with in vitro and in vivo experiments.Methods: R language was used to analyze middle cerebral artery occlusion/reperfusion (MCAO/R) rat transcriptome data and to identify differential gene expression following THSWD treatment. Gene set enrichment analysis (GSEA) was used to analyze the gene set enrichment pathway of MCAO/R rats treated with THSWD. PPI networks screened key targets. The Human Brain Microvascular Endothelial Cells (HBMEC) Oxygen Glucose Deprivation/Reoxygenation (OGD/R) model and SD rat models of MCAO/R were established. FITC-dextran, immunofluorescence, flow cytometry, ELISA, immunohistochemistry, Western blotting, and RT-qPCR were performed to identify potential treatment targets.Results: A total of 515 differentially expressed genes of THSWD in MCAO/R rats were screened and 92 differentially expressed genes of THSWD potentially involved in stroke intervention were identified, including Cd68, Ccl2, and other key genes. In vitro, THSWD reversed the increase in permeability of HBMEC cells and M1/M2 polarization of macrophages induced by CCL2/CCR2 axis agonists. In vivo, THSWD improved nerve function injury and blood-brain barrier injury in MCAO/R rats. Further, THSWD inhibited the infiltration and polarization of macrophages, reduced the expression of IL-6, TNF-α, and MMP-9, and increased the expression of IL-4, while reducing the gene and protein expression of CCL2 and CCR2.Conclusion: THSWD may play a protective role in ischemic stroke by inhibiting the CCL2/CCR2 axis, reducing the infiltration of macrophages, and promoting the polarization of M2 macrophages, thereby reducing inflammatory damage, and protecting injury to the blood-brain barrier.

Background: The causes of acute liver injury (ALI) are complex and diverse, including alcohol, drugs, infections, and toxic substances, and it has become a major global health issue. Traditional Chinese medicine (TCM), with advantages such as broad-spectrum biological activity, low toxicity, and minimal side effects, has been widely used in drug research and development as well as disease management. Some TCMs have shown significant efficacy in treating ALI: Ganoderma lucidum polysaccharides, monoterpene glycosides from Paeonia lactiflora, glycyrrhizic acid saponins, and flavonoids all exhibit liver-protective effects; however, the protective effects of their compound preparations on liver injury remain unclear.Methods: The study optimized the water extraction process using orthogonal experiments with AHP-entropy weight scoring. The preparation process was verified by single-factor experiments, Plackeett–Burman and Box–Bohnken tests. The mechanism of action was validated using network pharmacology methods and a CCl4-induced acute liver injury animal model.Results: The developed extraction and granule formation processes were reliable after validation. The contents of active components in CEHG determined by HPLC were as follows: albiflorin 1.26%, paeoniflorin 5.42%, liquiritin 0.43%, glycyrrhizic acid 1.30%, and ganoderic acid A 0.14%, with batch-to-batch variation coefficients (RSD) of 1.66%, 0.87%, 2.32%, 1.60%, and 4.03%, respectively. The dry extract ratio was 18.23% with an RSD of 2.11%. Network pharmacology revealed that CEHG improved liver injury by regulating the HIF-1, p53, and FoxO signaling pathways. Animal experiments indicated that CEHG granules reduce liver function-related aspartate transaminase (AST), alanine transaminase (ALT), total cholesterol (TC), triglyceride (TG), total bilirubin (TBIL), and LDH levels, decrease the levels of inflammatory factors IL-6, IL-1β, and TNF-α and of oxidative stress-related MDA and ROS, and down-regulated the mRNA and protein levels of P53, Bax, and HIF-1α in rats with liver injury. Meanwhile, CEHG improved liver function-related Total Protein (TP), raised the levels of oxidative stress-related SOD, GSH-Px, and GSH, and up-regulated mRNA and protein expression of Bcl-2.Conclusion: This study successfully optimized the extraction and granule-formation of CEHG and revealed its hepatoprotective mechanism through network pharmacology and animal studies, providing scientific evidence supporting the further development and clinical use of CEHG.

In our previous studies, we have identified that a series of message RNAs (mRNAs), microRNAs (miRNAs) and long noncoding RNAs (lncRNAs) may contribute to stroke. However, the network of RNA regulation with stroke remains well-known enough. This study explored the regulatory mechanism and function of lncRNAs as ceRNAs in stroke. According to the competitive endogenous RNA(ceRNA) hypothesis, a group of lncRNA, miRNA and mRNAs were found, which revealed the interaction between lncRNA, miRNA and mRNA and stroke. Several important lncRNAs were selected for in-depth analysis including sub-ceRNA-network, GO enrichment analyis, and pathway enrichment analysis. Importantly, the candidate lncRNAs, miRNAs and mRNAs are contributed to the ceRNA network, which can be used as potential therapeutic targets for stroke for further research.