Figure 1

pri-miR-9-BMSCs could attenuate SAP. (A) BMSCs infected by pri-miR-9- and Empty- virus were expressing the Green Fluorescent Protein (GFP). (B) The identification of recombinant pri-miR-9-1-PCDH-CMV-MSCs-EF1-GFP-T2A-Puro plasmid (pri-miR-9-1-PCDH) by double enzyme digestion and a 368 bp DNA fragment (pri-miR-9-1) was shown as red arrow. (C) pri-miR-9-1 was amplified from pri-miR-9-1-PCDH by applying the special primers. (D and E) The expression of mature miR-9 in pri-miR-9-BMSCs was higher than that in Empty virus-BMSCs by gPCR and qRT-PCR. Data are shown as mean ± SD for at least 3 separate experiments. ^###p < 0.001, compared with Empty virus-BMSCs by paired t test. (F,H,K,G) pri-miR-9-BMSCs could markedly reduce the pancreatic edema, infiltration, hemorrhage and necrosis, decrease the levels of serum amylase, lipase and pro-inflammatory cytokines (IL-1β, IL-6, TNF-α, HMBG1, and MPO) and increase the levels of serum anti-inflammatory cytokines (IL-4, IL-10, and TGF-β), compared with SAP, SAP+PBS, BMSCs, Empty virus-BMSCs, or TuD-BMSCs groups. (M,W) The cell apoptosis was significantly reduced by pri-miR-9-BMSCs, compared with SAP, SAP+PBS, BMSCs, Empty virus-BMSCs, or TuD-BMSCs groups. (P and Q) The expression of miR-9 in damaged pancreas was significantly increased by pri-miR-9-BMSCs, compared with NC, SAP, SAP+PBS, BMSCs, or Empty virus-BMSCs groups. Data are shown as mean ± SD for at least 3 separate experiments. ^%P < 0.05, ^%%p < 0.01, ^%%%p < 0.001, compared with NC, ^**p < 0.01 and ^***p < 0.001, compared with NC, ^@@p < 0.01 and ^@@@p < 0.001, compared with SAP, ^&&p < 0.01 and ^&&&p < 0.001, compared with PBS treatment (SAP+PBS), ^#p < 0.05, ^##p < 0.01 and ^###p < 0.001, compared with BMSCs, ^$p < 0.05, ^$$p < 0.01 and ^$$$p < 0.001, compared with Empty virus-BMSCs, ⁺p < 0.05 and ⁺⁺⁺p < 0.001, compared with TuD-BMSCs by using two-tailed t test. (X) GFP-BMSCs could deliver exogenous Cy3-miR-9a-5p to the liver, spleen, lung and pancreas of SAP rats, of which the number was observed more in the liver and spleen. gPCR, General PCR, qRT-PCR, quantitative Real-time PCR, SAP, severe acute pancreatitis.