动物造模,肾纤维化 Z-bio 2023-03-20 322

      Objective To investigate the expression of miR-146b-5p in a mouse model of renal fibrosis, and to investigate the effect of knocking down miR-146b-5p in vivo on renal injury and fibrosis in mice.

　　Methods 24 8-week-old C57BL/6 male mice were randomly divided into sham operation group (sham), UUO model group (UUO), and UUO+renal miR-146b-5p electroknockdown group (UUO-KD), with 8 mice in each group. In the Sham group, only skin incision was performed to expose and dissociate the right renal ureter without ligation or disconnection. Animal models of unilateral ureteral obstruction (UUO) were established in the UUO group. In the UUO-KD group, a specific miR-146b-5p knockdown was performed on the mouse kidney by electroporation of the CRISPR/RfxCas13 d plasmid. After 24 hours, a UUO mouse model was established according to the model group method. After 7 days, the mice were killed to collect kidney samples. HE staining was used to observe renal pathological changes, Masson to detect the degree of renal interstitial fibrosis, and immunohistochemistry to detect fibrosis related proteins（ α- SMA, FN, Col-1) expression, Western Blot, Real-time PCR detection of miR-146b-5p α- SMA、FN、IL-1 β、 IL-6、TNF- α Changes in isogenes.

　　Results miR 146b-5p significantly increased in the UUO model, and the gene significantly decreased after knocking down miR 146b-5p by electroporation (P<0.05) 05), while IL-1, IL-6, TNF- α The expression of inflammatory factors was significantly decreased (P<0.01) 05) After HE and Masson staining, it was observed that the renal structure in the UUO-KD group was better than that in the UUO group, with slight tubular deformation, and significant improvement in renal injury and fibrosis. Immunohistochemical results showed that: α- Fibrosis indicators such as SMA, FN, and Col-1 were also significantly decreased in the UUO-KD group (P<0.01) 0001)。

　　Conclusion Inhibiting the overexpression of miR-146b-5p in UUO can significantly improve renal fibrosis, and miR-146b-5p may be a potential therapeutic target for renal fibrosis.