M2 macrophage-derived exosomal miR-145-5p protects against the hypoxia/reoxygenation-induced pyroptosis of cardiomyocytes by inhibiting TLR4 expression

Abstract

Background: Exosomes carrying micro ribonucleic acids (miRNAs) protect against myocardial ischemic injury. In the study, we sought to investigate the protective effect mechanism of M2 macrophage-derived exosome miR-145-5p in hypoxia-reoxygenation (H/R)-induced cardiomyocytes.Methods: M2 macrophages were isolated and induced from blood donated by healthy donors. M2 macrophages were transfected with or without miR-145-5p. Exosomes derived from M2 macrophages were isolated and identified by flow cytometry, nanoparticle tracking analysis, and transmission electron microscopy (TEM). AC16 cells were used to establish an H/R model, and cell activity was detected using a Cell Counting Kit 8 (CCK-8). Western blot was used to detect the expression of gasdermin D (GSDMD), nucleotide-binding domain-like receptor protein 3 (NLRP3), and caspase-1 in the H/R-induced AC16 cells to evaluate pyroptosis.Immunofluorescence staining was used to detect the positive rates of PKH26 and caspase-1. Combined with database prediction, dual luciferase reporter assays were used to validate toll-like receptor 4 (TLR4) as a downstream target molecule of miR-145-5p. A real-time quantitative polymerase chain reaction (RT-qPCR) analysis and western blot were used to detect the expression of TLR4 in the AC16 cells.Results: Flow cytometry, western blot, nanoparticle tracking and TEM results confirmed the successful isolation of M2 macrophage-derived exosomes. CCK-8 results showed M2 macrophage-derived exosomes decreased the viability of the H/R-induced cells. Western blot results showed the expressions of GSDMD, caspase-1, and NLRP3 were significantly downregulated in the H/R group. Moreover, CCK-8 results showed the M2 macrophage-derived exosome miR-145-5p significantly ameliorated H/R-induced AC16 cellular activity. Western blot results confirmed the expressions of GSDMD, NLRP3, and caspase-1 were significantly downregulated in the macrophage-derived exosome miR-145-5p group compared to the M2 macrophage-derived exosome NC (normal control) group. Immunofluorescence staining results displayed the same trend in terms of the caspase-1 positivity rate. Further, we demonstrated overexpression of TLR4 partially reversed the protective effect of M2 macrophage-derived exosome miR-145-5p in the H/R-induced AC16 cells. Additionally, overexpression of TLR4 reversed the protein expression associated with pyroptosis in M2 macrophage-derived exosome miR-145-5p in the H/R-induced AC16 cells.Conclusions: Our study indicated M2 macrophage-derived exosomes carrying miR-145-5p inhibited H/ R-induced cardiomyocyte pyroptosis by downregulating the expression of TLR4.

Advanced Nanoarchitectonics of Drug Delivery Systems with Pyroptosis Inhibition for Noncancerous Disease Treatment

“…Reduced the activation of caspase-1 PFTU@DEX NPs [ 109] Dexamethasone Acute lung injury LPS-induced ALI mice model Inhibited ROS-NLRP3 pathway NM-LaCD NPs [ 52] 𝛼-cyclodextrin Neutrophilic asthma Neutrophilic asthma mice Attenuate the NETs-mediated inflammasome activation DNA nano-prism [ 97] Buformin/p65 siRNA Acute lung injury LPS-induced ALI mice model Inhibiting the NF𝜅B p65/NLRP3 pathway Disease of cardiovascular system M2 macrophage-derived exosome [ 110] MicroRNA-145-5p Myocardial ischemia/reperfusion injury Hypoxia/reoxygenation AC16 cells model Downregulate the TLR4-NLRP3 pathway PDA-based biomimetic nanoplatform [ 111] Polydopamine Myocardial Ischemia/reperfusion injury MI/RI rat model Suppressing the NLRP3-caspase-1 pathway Calcium carbonate nanoarchitectonics [ 76] Colchicine Acute myocardial infarction MI rat Model Suppressing TLR4/NF𝜅B/NLRP3 pathway…”

“…[120] Using the advantages of exosomes, Li et al harnessed exosomes derived from M2 macrophages to deliver miRNA-145-5p. [110] This approach resulted in the development of a naturally occurring nanomaterial with low immunogenicity and low toxicity, which effectively suppressed myocardial cell apoptosis induced by H/R by downregulating the expression of TLR4. Compared to free drugs, nanoarchitectonics-based drug delivery systems exhibit a prolonged circulation time in the bloodstream, which allows for enhanced accumulation of drugs at the target site, consequently improving therapeutic efficacy.…”

Adipose tissue macrophage-derived exosomal miR-210-5p in modulating insulin sensitivity in rats born small for gestational age with catch-up growth

“…MicroRNAs (miRNAs) are a class of highly conserved, endogenous, single-stranded noncoding small RNAs that regulate gene expression by binding to the 3' untranslated region (UTR) of target mRNA, causing translational arrest or degradation ( 14 ). Emerging evidence has revealed that exosomal miRNAs play a crucial role in insulin resistance and glucose intolerance in both animals and humans ( 15 , 16 ). For example, adipose tissue macrophage (ATM)–derived exosomal miR-155 can cause insulin resistance in vivo and in vitro ( 17 ).…”

Enhanced Osteolysis Targeted Therapy through Fusion of Exosomes Derived from M2 Macrophages and Bone Marrow Mesenchymal Stem Cells: Modulating Macrophage Polarization

“…[21] Concurrent studies corroborate the capacity of M2-Exos to modulate macrophage polarization and temper inflammation. [22,23] Considering the feasibility of biofilm fusion and exosomal liposome fusion, along with the regulatory influence wielded by M2-Exos and BMSCs-Exos on macrophage polarization, a novel milestone has been achieved through the creation of M2-Exos and BMSCs-Exos fused exosomes (M2-BMSCs-Exos). This groundbreaking achievement seamlessly amalgamates immune resistance with precise targeting of osteolysis sites, effectively harnessing the therapeutic potential of two distinct exosome types.…”