C57BL/6亚品系在多柔比星诱导FSGS模型中的差异研究OA

Objective To investigate the optimal protocol for a doxorubicin-induced chronic kidney disease mouse model used for studying human focal segmental glomerulosclerosis(FSGS).Methods C57BL/6J and C57BL/6N mice were randomly assigned using a random number sequence into control groups(J0 and N0,24 mice each)and FSGS model groups(J1,J2,N1,and N2,24 mice each).J1 and N1 groups received a single tail vein injection of 15 mg/kg doxorubicin,J2 and N2 groups received two injections(with a two-week interval),and J0 and N0 groups received an equal volume of 0.9%sodium chloride injection.The overall condition and body weight changes of the mice were observed,and general data were recorded.Urine was collected to evaluate proteinuria through urinary protein and urinary creatinine.Kidney tissue proteins were extracted for Western blot analysis to detect the expression of podocyte marker proteins.Kidney histopathological manifestations were observed using hematoxylin-eosin staining,periodic acid-Schiff staining and transmission electron microscopy.Results After modeling,C57BL/6N mice showed worse general condition and more significant weight loss than C57BL/6J mice(P<0.001).In both strains,mice receiving two doxorubicin injections showed poorer general condition and more pronounced weight loss(P<0.001).The urinary protein-to-creatinine ratio in the doxorubicin-injected groups of both C57BL/6J and C57BL/6N was higher than that in the control groups at all time points(P<0.05).In the single-injection C57BL/6J group,the expression of some podocyte marker proteins[synaptopodin,Wilms tumor protein 1(WT1)]showed no significant changes,while in the two-injection group,the expression levels of podocyte marker proteins(nephrin,synaptopodin,podocalyxin,WT1,α-tubulin,podocin)were all lower than those in the control group.Both single-and two-injection C57BL/6N groups showed lower expression level of podocyte marker proteins than that in the control group,with more significant decreases in the two-injection group(P<0.05).Light and electron microscopy both confirmed successful modeling in the single-and two-injection C57BL/6N groups,with more significant pathological changes in the two-injection group.Conclusions In the doxorubicin-induced FSGS model in C57BL/6 mice,the C57BL/6N substrain outperforms C57BL/6J substrain in modeling efficacy.The recommended optimal modeling protocol for FSGS is the single tail vein injection of 15 mg/kg doxorubicin in C57BL/6N mice.