动物造模,小鼠肾纤维化 中国实验动物信息网 2023-06-12 802

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

　　Methods Twenty four eight week old C57BL/6 male mice were randomly divided into a Sham surgery group (sham), a UUO model group (UUO), and a UUO+renal miR-146b-5p electroporation knockdown group (UUO-KD) with 8 mice in each group. The Sham group only underwent skin incision, exposing and dissociating the right renal ureter without ligation or disconnection. Unilateral ureteral obstruction (UUO) animal model was established in the UUO group. The UUO-KD group underwent specific miR-146b-5p knockdown in the mouse kidneys by first electroporating the CRISPR/RfxCas13 d plasmid. After 24 hours, a UUO mouse model was established using the model group method. Seven days later, the mice were euthanized and kidney specimens were collected. HE staining was used to observe renal pathological changes, Masson was used to detect the degree of renal interstitial fibrosis, and immunohistochemistry was used 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.

　　The results showed that miR -146b-5p significantly increased in the UUO model, and after knocking down miR -146b-5p by electroporation, the gene significantly decreased (P<0.05) 05), at the same time, IL-1, IL-6, TNF- α The expression of inflammatory factors was significantly downregulated (P<0.05) 05) After HE and Masson staining, it was observed that the UUO-KD group had a better renal structure compared to the UUO group, with slight tubular deformation, and significant improvement in renal injury and fibrosis. And the immunohistochemical results showed that: α- Fibrosis indicators such as SMA, FN, and Col-1 were also significantly reduced in the UUO-KD group (P<0.05) 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.