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MicroRNAs (miRNAs) are a novel class of powerful, endogenous regulators of gene expression. This study identified 16 differentially expressed miRNAs in ischemic myocardium of rats using TaqMan Low Density Array. In addition, bioinformatics analyses, such as Gene ontology and Pathway assays, were applied to determine the apoptosis pathway, only regulated by miR-384-5p, and all the associated target genes (PIK3CD, PPP3CA, PPP3CB, PPP3R1, CASP3 and IL1A). These target genes, besides PIK3CB, were shown to be significantly up-regulated by qRT-PCR assay, which further suggested that PIK3CD, PPP3CA, PPP3R1, CASP3, IL1A could be regulated by miR-384-5p. MTT, Western blot, qRT-PCR and luciferase assays were used to investigate the role of miR-384-5p in myocardial ischemia. We found that cleaved caspase3 expression was up-regulated by miR-384-5p and down-regulated by miR-384-5p inhibitor suggesting that apoptosis pathway was regulated by miR-384-5p. We also found that miR-384-5p suppressed cell viability while miR-384-5p inhibitor improved it, confirming H9c2 cell survival was affected by miR-384-5p. In addition, the PIK3CD protein level in H9c2 cells was up-regulated by miR-384-5p inhibitor. We found that miR-384-5p expression level decreased and PIK3CD protein level increased in both ischemic myocardium of rats and hypoxic H9c2 cells, and that miR-384-5p suppress PIK3CD expression through a miR-384-5p binding site within the 3' untranslational region of PIK3CD. These results show that miR-384-5p, an important protecting factor, plays a significant role in cardioprotection by regulating PIK3CD in myocardial ischemia.

• A combined strategy of network pharmacology, transcriptomics, proteomics and molecular biology was used to study the active ingredients and mechanisms of TCM. • A network pharmacology approach based on the active ingredients of TCM absorbed into the brain was first proposed. • The pharmacodynamics of YQTL on CIRI with different period were comprehensively evaluated. • The active ingredients and mechanisms of YQTL was preliminarily clarified. • 54 direct targets and 28 indirect targets of YQTL against CIRI were preliminarily clarified. Ischemic stroke is caused by local lesions of the central nervous system and is a severe cerebrovascular disease. A traditional Chinese medicine, Yiqi Tongluo Granule (YQTL), shows valuable therapeutic effects. However, the substances and mechanisms remain unclear. We combined network pharmacology, multi-omics, and molecular biology to elucidate the mechanisms by which YQTL protects against CIRI. We innovatively created a combined strategy of network pharmacology, transcriptomics, proteomics and molecular biology to study the active ingredients and mechanisms of YQTL. We performed a network pharmacology study of active ingredients absorbed by the brain to explore the targets, biological processes and pathways of YQTL against CIRI. We also conducted further mechanistic analyses at the gene and protein levels using transcriptomics, proteomics, and molecular biology techniques. YQTL significantly decreased the infarction volume percentage and improved the neurological function of mice with CIRI, inhibited hippocampal neuronal death, and suppressed apoptosis. Fifteen active ingredients of YQTL were detected in the brains of rats. Network pharmacology combined with multi-omics revealed that the 15 ingredients regulated 19 pathways via 82 targets. Further analysis suggested that YQTL protected against CIRI via the PI3K-Akt signaling pathway, MAPK signaling pathway, and cAMP signaling pathway. We confirmed that YQTL protected against CIRI by inhibiting nerve cell apoptosis enhanced by the PI3K-Akt signaling pathway. [Display omitted] [ABSTRACT FROM AUTHOR]

Background: Breast cancer is still the most common malignant tumor that threatens the female's life in the world, especially triple-negative breast cancer (TNBC), one of the most difficult subtypes. Lack of targeted therapies brings about urgent demand for novel treatments. In this study we aim to investigate the anti-tumor activity of Berberine (BBR), a Chinese plant-derived alkaloid, against the TNBC cell line MDA-MB-231 and elucidate its mechanism referring to anti-inflammation. Methods: Cell inhibition rate was measured by Cell Proliferation Assay, the cytotoxic effects was detected by Lactate dehydrogenase (LDH) leakage assay, the colony formation and migration potential were evaluated by colony formation assay and wound healing assay, the release of inflammatory cytokines was detected by EMD multifactor detection, and alterations of proteins and genes related to the NLR family pyrin domain containing 3 (NLRP3) inflammasome pathway were analyzed using western blotting and real-time Polymerase Chain Reaction (PCR). Results: BBR reduce the viability of MDA-MB-231 cells and increased the release of LDH from the cells in a dose-dependent manner, with and inhibition of colony formation potential and migration of the cells. BBR also caused a marked reduction in the secretion of proinflammatory cytokines, Interleukin-1α (IL-1α), Interleukin-1β (IL-1β), Interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). Besides, a down-regulated behavior was observed with the expression of P2X purinoceptor 7 (P2X7), NLRP3, pro-caspase-1, apoptosis-associated speck-like protein containing a caspase-activation and recruitment domain (ASC), caspase-1 p20, Interleukin-18 (IL-18), IL-1β proteins and NLRP3, Caspase-1 and ASC mRNAs in the NLRP3 inflammasome cascade. Conclusions: Our results confirmed that BBR can effectively affect both tumor outgrowth and spontaneous metastasis in TNBC, and that we identified a new mechanism associated with inhibition the NLRP3 inflammasome pathway, suggesting its potential therapeutic relevance in clinical use. [ABSTRACT FROM AUTHOR]