Exaggerated Renal Fibrosis In LncRNA Gas5-deficient Mice After Unilateral Ureteric Obstruction

Objective:This study set out to assess the effect of long non-coding RNA growth arrest-specific 5（lncRNA Gas5）in renal fibrosis induced by operation of unilateral ureteric obstruction（UUO）in mice.Methods:12 Gas5^+/-mice were constructed by CRISPR/Cas9technology.Meanwhile 12 wild-type mice（WT）were selected.Gas5^+/-and WT mice were subjected to either UUO or sham operation.The mice were sacrificed 10 Days after the operation,and the plasma and operative kidney were taken for various analyses.Blood plasma were obtained from the mice for the measurement of blood urea nitrogen（BUN）,uric acid（UA）,serum creatinine（SCr）,high sensitivity C reactive protein（hs CRP）,and urine albumin-to-creatinine ratio（UACR）.Masson staining was performed to observe the degree of collagen deposition,and immunohistochemical staining was performed to detect the distribution and expression ofα-smooth muscle actin（α-SMA）,vimentin,collagenⅣand fibronectin in the extracellular matrix of kidney.q RT-PCR and Western blotting was performed to detect the m RNA and protein expression ofα-SMA,vimentin,collagenⅣ,E-cadherin,transforming growth factor-β（TGF-β）,phosphatase and tensin homologue（PTEN）and matrix metalloprotease-9（MMP-9）.Results:Unlike the wild type mice,the obstructed kidney in Gas5^+/-mice showed more severe renal fibrosis and collagen deposition.In the UUO-Gas5^+/-group,the serum levels of uric acid,serum creatinine,and the urine levels of albumin-to-creatinine ratio were higher.Moreover,the expression of m RNA and protein ofα-smooth muscle actin（α-SMA）,vimentin,collagenⅣ,fibronectin,and matrix metalloproteinase 9（MMP-9）were higher,whereas that of phosphatase and tensin homolog（PTEN）were lower with the difference being statistically significant（p<0.05）.Conclusion:lncRNA Gas5 was up-regulated in renal fibrosis tissues,and its deficiency exacerbated renal fibrosis in the UUO mice model.