Preparation of kidney-specific ALR knockout mice and the effect of ALR on renal tubulointerstitial fibrosis
10.13431/j.cnki.immunol.j.20240094
- VernacularTitle:肾脏特异性ALR基因敲除小鼠模型的制备及ALR对肾纤维化的影响
- Author:
Lili HUANG
1
;
Yixin MA
;
Fangyan TAN
;
Pengfei YANG
;
Qi LIU
;
Ling ZHANG
;
Xiaohui LIAO
Author Information
1. 400010,重庆医科大学附属第二医院肾内科
- Publication Type:Journal Article
- Keywords:
ALR;
Kidney gene knockout;
Kidney fibrosis;
Macrophage polarization
- From:
Immunological Journal
2024;40(8):679-685
- CountryChina
- Language:Chinese
-
Abstract:
Objective To generate a stable kidney-specific deletion of augmenter of liver regeneration(ALR)mice and provide an animal model for further studying the biological function of ALR in the kidney.In addition,we further explored the effect of ALR on renal tubulointerstitial fibrosis.Methods Mating and identification of ALRflox/+/Ggt1-Cre mice with ALRflox/flox mice was carried out,and the ALRflox/flox/Ggt1-Cre mice were screened.Mating and identification of ALRflox/flox/Ggt1-Cre mice with ALRflox/flox mice were carried out,and screening the ALRflox/flox/Ggt1-Cre mice was performed.The ALRflox/flox/Ggt1-Cre mouse was a kidney-specific ALR knockout mouse and the ALRflox/flox mouse was a control mouse.The mouse genotypes were identified by PCR;the expression of ALR protein and mRNA levels in mouse liver were detected by Western blotting and real-time PCR;ALR expression in mouse kidney was detected by immunofluorescence.The kidney tissue morphology of the mouse kidney was observed using hematoxylin-eosin staining,periodic acid Schiff's reaction and Masson stain.Flow cytometry,immunohistochemistry,immunofluorescence and real time PCR analyses were used to determine macrophage phenotype.Results PCR results indicate that mouse genotypes are consistent with ALRflox/flox/Ggt1-Cre.Compared with ALRflox/flox mice,ALRflox/flox/Ggt1-Cre mice showed lower levels of ALR mRNA and protein,more interstitial inflammatory cells in kidney tissue,broken basement membrane of the renal tubules,the brush-border damage and significant interstitial fibrosis.After the kidney-specific ALR gene was knocked out,macrophages infiltrated the kidney tissue o and the polarization process of macrophages from M1 to M2 was reduced.Conclusion In this study,the kidney-specific deletion of ALR mouse was successfully constructed using the Cre/Loxp technology,which providing an important animal model for further study of the role of ALR genes.And ALR deficiency promotes renal fibrosis in physiological conditions.
