Various fish species in China's aquaculture industry are impacted by hemorrhagic disease, the culprit being Grass carp reovirus genotype (GCRV). In spite of extensive research, the causative factors behind GCRV's disease development are poorly understood. A rare minnow is an exemplary model system for scrutinizing the development of GCRV disease. Metabolic changes in the spleen and hepatopancreas of rare minnows injected with virulent GCRV isolate DY197 and attenuated isolate QJ205 were investigated using liquid chromatography-tandem mass spectrometry metabolomics. Metabolic profiling after GCRV infection indicated changes in both the spleen and hepatopancreas, where the more aggressive DY197 strain displayed a more marked variation in metabolites (SDMs) than the attenuated QJ205 strain. Besides this, most SDMs displayed a diminished expression in the spleen, in contrast to an enhanced expression in the hepatopancreas. Kyoto Encyclopedia of Genes and Genomes pathway analysis revealed tissue-specific metabolic adjustments in response to viral infection. The virulent DY197 strain induced more amino acid metabolism pathways in the spleen, particularly tryptophan, cysteine, and methionine pathways, essential for the host's immune response. Simultaneously, both virulent and attenuated viral strains enhanced nucleotide metabolism, protein synthesis, and associated pathways within the hepatopancreas. The study of rare minnow metabolism in response to variable GCRV infections, from attenuated to virulent, will significantly improve our comprehension of viral pathogenesis and host-pathogen interactions.
China's southern coastal aquaculture industry centers on the humpback grouper, Cromileptes altivelis, because of its notable economic contribution. Toll-like receptor 9 (TLR9), a key player within the toll-like receptor family, identifies unmethylated CpG motifs in oligodeoxynucleotides (CpG ODNs) originating from bacterial and viral genomes, thereby functioning as a pattern recognition receptor to activate the host immune system. The C. altivelis TLR9 (CaTLR9) ligand CpG ODN 1668 was found to substantially enhance the antibacterial immunity of humpback grouper, both in living specimens and in cultured head kidney lymphocytes (HKLs) in vitro. Furthermore, CpG ODN 1668 additionally fostered the growth of cells and upregulated immune gene expression in HKLs, while also fortifying the phagocytic capabilities of head kidney macrophages. Silencing CaTLR9 expression within the humpback group resulted in a considerable decrease in the expression levels of TLR9, MyD88, TNF-, IFN-, IL-1, IL-6, and IL-8, consequently abolishing most of the antibacterial immune effects attributable to CpG ODN 1668. In conclusion, CpG ODN 1668's ability to induce antibacterial immune responses was fundamentally linked to the CaTLR9-dependent pathway. These outcomes illuminate the antibacterial immune responses within fish TLR signaling pathways, underscoring the potential of this research for the discovery of natural antibacterial compounds from fish.
Remarkably tenacious, Marsdenia tenacissima (Roxb.) exhibits an enduring nature. Wight et Arn. constitutes a facet of traditional Chinese medicine. Xiao-Ai-Ping injection, a standardized extract (MTE), is widely employed in the treatment of cancer. MTE's pharmacological impact on cancer cells, leading to their demise, has been a subject of detailed study. Nonetheless, the question of whether MTE initiates tumor endoplasmic reticulum stress (ERS)-associated immunogenic cell death (ICD) remains unanswered.
To investigate the possible role of endoplasmic reticulum stress in the anti-cancer effects of MTE, and to identify potential mechanisms of endoplasmic reticulum stress-induced immunogenic cell death by MTE.
The influence of MTE on tumor growth inhibition in non-small cell lung cancer (NSCLC) was assessed using CCK-8 and a wound healing assay. Network pharmacology analysis, in conjunction with RNA sequencing (RNA-seq), was undertaken to verify the biological modifications within NSCLC cells following treatment with MTE. Our analysis of endoplasmic reticulum stress relied on Western blot, qRT-PCR, reactive oxygen species (ROS) assay, and mitochondrial membrane potential (MMP) assay. The immunogenic cell death-related markers were studied using ELISA in conjunction with an ATP release assay. Salubrinal played a role in inhibiting the endoplasmic reticulum stress response mechanism. To hinder AXL's activity, siRNAs and bemcentinib (R428) were utilized. Following treatment with recombinant human Gas6 protein (rhGas6), AXL phosphorylation returned. MTE's effect on endoplasmic reticulum stress and the immunogenic cell death response was unequivocally proven through in vivo models. The AXL inhibiting compound from MTE was explored by molecular docking, and its effect was further confirmed by means of Western blot analysis.
MTE's presence led to a reduction in the viability and migratory abilities of PC-9 and H1975 cells. Post-MTE treatment, the enrichment analysis showcased a pronounced enrichment of differential genes directly involved in endoplasmic reticulum stress-related biological processes. Subsequent to MTE administration, a decrease in mitochondrial membrane potential (MMP) and an increase in ROS levels were detected. Subsequent to MTE treatment, endoplasmic reticulum stress-related proteins (ATF6, GRP-78, ATF4, XBP1s, and CHOP) and immunogenic cell death markers (ATP, HMGB1) displayed increased expression, and AXL phosphorylation was correspondingly decreased. Simultaneous exposure of cells to salubrinal, an endoplasmic reticulum stress inhibitor, and MTE caused a diminished suppression of PC-9 and H1975 cells by MTE. Fundamentally, curtailing AXL expression or activity also prompts the expression of markers signifying both endoplasmic reticulum stress and immunogenic cell death. MTE's mechanistic action resulted in suppressed AXL activity, inducing endoplasmic reticulum stress and immunogenic cell death; this effect lessened when AXL activity was re-established. Correspondingly, MTE substantially increased the expression of endoplasmic reticulum stress-related indicators in the tumor tissues of LLC-bearing mice, and correspondingly elevated plasma levels of ATP and HMGB1. The molecular docking studies indicated that kaempferol possesses the strongest binding energy with AXL, thus inhibiting AXL phosphorylation activity.
MTE's action results in endoplasmic reticulum stress and subsequent immunogenic cell death within NSCLC cells. For the anti-tumor activity of MTE to manifest, endoplasmic reticulum stress must be present. AXL activity is suppressed by MTE, thereby triggering endoplasmic reticulum stress-associated immunogenic cell death. serious infections In MTE, kaempferol acts as an active inhibitor of AXL activity. This study uncovered AXL's function in modulating endoplasmic reticulum stress, expanding the anti-tumor strategies of MTE. Additionally, kaempferol has the potential to be considered a novel substance that inhibits AXL.
The induction of endoplasmic reticulum stress-associated immunogenic cell death in NSCLC cells is a consequence of MTE. Anti-tumor effects of MTE are contingent on a stimulated endoplasmic reticulum stress response. selleckchem The inhibition of AXL activity by MTE is a crucial step in triggering endoplasmic reticulum stress-associated immunogenic cell death. MTE cells experience a suppression of AXL activity due to the active component, kaempferol. The current investigation uncovered the function of AXL in modulating endoplasmic reticulum stress, thus augmenting the anti-tumor effects of MTE. Beyond that, kaempferol is potentially a novel inhibitor targeting the AXL receptor.
Individuals with chronic kidney disease stages 3 through 5 develop complications in their skeletal systems, which are medically termed Chronic Kidney Disease-Mineral Bone Disorder (CKD-MBD). This condition has a strong correlation with an elevated rate of cardiovascular diseases and a critical impact on patients' quality of life. In the realm of traditional Chinese medicine for treating CKD-MBD, salt Eucommiae cortex, featuring its kidney-tonifying and bone-strengthening abilities, stands out in clinical application more so than Eucommiae cortex. However, the mechanics involved in its operation are still not clear.
This research integrated network pharmacology, transcriptomics, and metabolomics to examine the impacts and processes of salt Eucommiae cortex on CKD-MBD.
Eucommiae cortex salt was administered to CKD-MBD mice, which were generated by 5/6 nephrectomy and a low calcium/high phosphorus diet. Renal functions and bone injuries were diagnosed by means of serum biochemical detection, histopathological analysis, and femur Micro-CT imaging. androgen biosynthesis By analyzing transcriptomic data, differentially expressed genes (DEGs) were identified in comparisons between the control group and the model group, between the model group and the high-dose Eucommiae cortex group, and between the model group and the high-dose salt Eucommiae cortex group. The study employed metabolomics to analyze the differentially expressed metabolites (DEMs) comparing the control group to the model group, the model group to the high-dose Eucommiae cortex group, and the model group to the high-dose salt Eucommiae cortex group. Through an integrated approach employing transcriptomics, metabolomics, and network pharmacology, common targets and pathways were discovered and subsequently proven by in vivo experimentation.
Salt Eucommiae cortex therapy efficiently ameliorated the negative effects on the renal system and bone integrity. In comparison to CKD-MBD model mice, the serum BUN, Ca, and urine Upr levels were demonstrably lower in the salt Eucommiae cortex group. The integrated analysis of network pharmacology, transcriptomics, and metabolomics showcased Peroxisome Proliferative Activated Receptor, Gamma (PPARG) as the only shared target, primarily operating within AMPK signaling pathways. In CKD-MBD mice, kidney tissue PPARG activation displayed a pronounced decline, which was substantially counteracted by treatment with salt Eucommiae cortex.