Fig 2: MiR-219c-3p overexpression ameliorated acute kidney inflammation.A Schematic overview of study in AKI model induced by UUO mice with miR-219c-3p overexpression via lentiviral vector transfection. B Expression of miR-219c-3p in the UUO kidney transfected with miR-219c lentiviral or NC as control. C–D Representative sections of mice kidney stained for F4/80+ macrophages. The number of infiltrated macrophages were quantified. Scale bar, 100 µm. E–G Mincle, MCP-1, TNF-a was detected by RT-PCR in the kidney which were inhibited by miR-219c-3p overexpression. Data were presented as mean ± SD. *p < 0.05 compared to NC_UUO. NC_UUO, UUO mice treated with empty lentiviral vector as a negative control. MiR-219c lentiviral_UUO, UUO mice treated with miR-219c-3p overexpression via lentiviral transfection.

Fig 3: Overexpression of Mincle recovered A&P and Bifidobacterium containing serum inhibited inflammation in LPS + IS stimulated RAW264.7 cells. (A–C) Real-time PCR was performed to detect the mRNA expression of inflammatory indicators (IL-1ß, IL-6, TNF-a) in RAW264.7 cells after overexpression of Mincle; (D–F) ELISA was used to detect the secretion of inflammatory factors (IL-1ß, IL-6, TNF-a) in supernatant of RAW264.7 cells after overexpression of Mincle; (G,H) The mRNA expression of iNOS and Mincle in RAW264.7 cells; (I) The protein levels of Mincle/iNOS/p-P65/p-65 in RAW264.7 cells after overexpression of Mincle. *P < 0.05, **P < 0.01, ***P < 0.001, vs. LPS + IS + A&P + BB Group.

Fig 4: Screening for universal shMincle sequences in human and mouse macrophagesKnockdown efficiencies of three designed sequences against Mincle in (A and B) murine macrophage RAW264.7 and (C and D) human macrophage phorbol 12-myristate 13-acetate (PMA)-stimulated THP-1 under cancer secretome stimulation in vitro. siMincle #3 showed the best silencing for Mincle in both human and murine macrophages. (E and F) Map of constructed shMincle plasmid using si#3 and pSUPER.puro vector. Data represent mean ± SEM of 3 independent in vitro experiments. Statistical analysis based on ordinary one-way ANOVA. ****p < 0.0001, ***p < 0.001, **p < 0.01 versus NC; ####p < 0.0001, ###p < 0.001 versus NC+CM.

Fig 5: USMB-shMincle effectively blocks the growth of human non-small-cell lung carcinoma (NSCLC) xenografts in vivoMice were inoculated with 1 × 106 human lung cancer A549 cells subcutaneously and then received USMB-mediated tumor-specific delivery with 100 µL of mixture with EV (USMB-EV) or shMincle plasmid (50 µg, 100 µg, and 200 µg/mouse; USMB-shMincle) every 5 days as treatment. (A–C) Imaging of A549 human lung cancer-bearing mice and tumors, tumor weight, and tumor size. (D) Mincle expression in TAMs was highly suppressed in 100-µg and 200-µg dosages of USMB-shMincle treatment. Data represent mean ± SEM of 5 mice/group. Statistical analysis based on ordinary one-way ANOVA. ****p < 0.0001, ***p < 0.001 versus Control; ####p < 0.0001, ###p < 0.001, #p < 0.05 versus EV-USMB; @@@@p < 0.0001, @@@p < 0.001, @@p < 0.01 versus 50 µg shMincle; ^p < 0.05 versus 100 µg shMincle. Scale bars, 100 µm.