C57BL/6亚品系在多柔比星诱导FSGS模型中的差异研究

doi:10.16138/j.1673-6087.2026.02.04

内科理论与实践 ›› 2026, Vol. 21 ›› Issue (02): 124-131.doi: 10.16138/j.1673-6087.2026.02.04

C57BL/6亚品系在多柔比星诱导FSGS模型中的差异研究

孙羽斐¹^,^*(), 刘爽¹^,^*, 李慧凛²(), 蒋更如¹^,³()

^1. 上海交通大学医学院附属新华医院肾脏内科，上海　200092
^2. 上海市浦东新区公利医院肾脏内科，上海　200135
^3. 上海市罕见病中心，上海　200092

收稿日期:2025-03-31 修回日期:2026-04-02 出版日期:2026-04-25 发布日期:2026-06-15
通讯作者: 李慧凛 E-mail：medlhl@163.com; 蒋更如 E-mail：jianggengru@xinhuamed.com.cn
作者简介:作者贡献/Authors’ Contributions孙羽斐负责课题设计、分子及动物实验的实施、文稿撰写；刘爽负责课题设计、分子及动物实验的实施；李慧凛负责课题设计、文章撰写指导；蒋更如负责课题设计、文章撰写指导。
*：孙羽斐和刘爽为共同第一作者
基金资助:
国家自然科学基金(82170713);上海市卫生健康委员会科研项目(202040293);上海市浦东新区卫生健康系统医学领先人才培养计划(PWRl2024-01);上海市浦东新区卫生健康委员会学科建设计划(PWZzb2022-03)

Comparison of doxorubicin-induced FSGS models across C57BL/6 substrains

SUN Yufei¹^,^*(), LIU Shuang¹^,^*, LI Huilin²(), JIANG Gengru¹^,³()

^1. Department of Nephrology, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China
^2. Department of Nephrology, Shanghai Pudong Gongli Hospital, Shanghai 200135, China
^3. Shanghai Centre for Rare Disease, Shanghai 200092, China

Received:2025-03-31 Revised:2026-04-02 Online:2026-04-25 Published:2026-06-15

摘要/Abstract

摘要：

目的：探究用于人类局灶节段性肾小球硬化（focal segmental glomerulosclerosis，FSGS）研究的多柔比星诱导的小鼠慢性肾脏病模型的最佳方案。方法：将C57BL/6J和C57BL/6N小鼠分别按照随机数字序列进行随机分组，分为对照组（J0组和N0组，各24只）和FSGS造模组（J1组、J2组、N1组、N2组，各24只）。J1组和N1组小鼠尾静脉注射多柔比星15 mg/kg 1次，J2组和N2组注射2次（间隔2周），J0组和N0组注射等量0.9%氯化钠注射液。观察小鼠状态及体重变化，记录一般情况；收集尿液检测尿蛋白和尿肌酐，评估蛋白尿；提取肾组织蛋白行免疫印迹实验，检测足细胞标志蛋白表达。通过苏木精-伊红染色、过碘酸-希夫染色及透射电子显微镜观察各组小鼠肾组织病理表现。结果：C57BL/6N小鼠造模后一般情况较C57BL/6J小鼠差，体重下降更显著（P<0.001）。两种品系小鼠经2次注射多柔比星后，一般情况更差，体重下降更明显（P<0.001）。C57BL/6J和C57BL/6N多柔比星注射组各时间段尿蛋白/肌酐比值均高于对照组（P<0.05）。C57BL/6J单次注射组部分足细胞标志蛋白[突触足蛋白、肾母细胞瘤蛋白1（Wilms tumor protein 1，WT1）]表达无显著变化，2次注射组足细胞标志蛋白（肾病蛋白、突触足蛋白、足萼蛋白、WT1、α微管蛋白、足蛋白）表达量均较对照组降低。C57BL/6N单次及2次注射组足细胞标志蛋白表达量均较对照组降低，2次注射组下降更明显（P<0.05）。光镜和电镜均显示C57BL/6N单次及2次注射组造模成功，2次注射组病理改变更显著。结论：在多柔比星诱导的C57BL/6小鼠FSGS模型中，C57BL/6N品系造模效果优于C57BL/6J品系。推荐FSGS适宜造模方案为C57BL/6N小鼠尾静脉单次注射15 mg/kg多柔比星。

关键词: 局灶节段性肾小球硬化, 多柔比星, 足细胞, C57BL/6J, C57BL/6N

Abstract:

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.

Key words: focal segmental glomerulosclerosis, doxorubicin, podocyte, C57BL/6J, C57BL/6N

孙羽斐, 刘爽, 李慧凛, 蒋更如. C57BL/6亚品系在多柔比星诱导FSGS模型中的差异研究[J]. 内科理论与实践, 2026, 21(02): 124-131.

SUN Yufei, LIU Shuang, LI Huilin, JIANG Gengru. Comparison of doxorubicin-induced FSGS models across C57BL/6 substrains[J]. Journal of Internal Medicine Concepts & Practice, 2026, 21(02): 124-131.

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参考文献 36

[1]	Liu G, He L. Epigallocatechin-3-gallate attenuates adriamycin-induced focal segmental glomerulosclerosis via suppression of oxidant stress and apoptosis by targeting hypoxia-inducible factor-1α/ angiopoietin-like 4 pathway[J]. Pharmacology,2019,103(5-6):303-314. doi: 10.1159/000496799
[2]	Wei M, Qiu Z, Li H, et al. Integrating network pharmacology approach and experimental validation to reveal the alleviation of Shenkangning capsule on chronic nephritis[J]. J Ethnopharmacol,2022,299:115676. doi: 10.1016/j.jep.2022.115676
[3]	Qi H, Fu J, Luan J, et al. miR-150 inhibitor ameliorates adriamycin-induced focal segmental glomerulosclerosis[J]. Biochem Biophys Res Commun,2020,522(3):618-625. doi: 10.1016/j.bbrc.2019.11.096
[4]	Qiu Y, Zhou J, Zhang H, et al. Rhodojaponin Ⅱ attenuates kidney injury by regulating TGF-β1/Smad pathway in mice with adriamycin nephropathy[J]. J Ethnopharmacol,2019,243:112078. doi: 10.1016/j.jep.2019.112078
[5]	Hu SW, Wang YH, Huang JS, et al. The PDE5 inhibitor, vardenafil, ameliorates progressive pathological changes in a focal segmental glomerulosclerosis mouse model[J]. Life Sci,2022,309:120992. doi: 10.1016/j.lfs.2022.120992
[6]	de Mik SM, Hoogduijn MJ, de Bruin RW, et al. Pathophysiology and treatment of focal segmental glomerulosclerosis: the role of animal models[J]. BMC Nephrol,2013,14:74. doi: 10.1186/1471-2369-14-74
[7]	Yang JW, Dettmar AK, Kronbichler A, et al. Recent advances of animal model of focal segmental glomerulosclerosis[J]. Clin Exp Nephrol,2018,22(4):752-763. doi: 10.1007/s10157-018-1552-8
[8]	Weiss RB. The anthracyclines: will we ever find a better doxorubicin?[J]. Semin Oncol,1992,19(6):670-686.
[9]	Nemoto S, Kubota T, Ohno H. Metabolic differences and differentially expressed genes between C57BL/6J and C57BL/6N mice substrains[J]. PLoS One,2022,17(12):e0271651. doi: 10.1371/journal.pone.0271651
[10]	Mekada K, Yoshiki A. Substrains matter in phenotyping of C57BL/6 mice[J]. Exp Anim,2021,70(2):145-160. doi: 10.1538/expanim.20-0158
[11]	Watanabe M, Kakutani M, Hiura K, et al. Differences in susceptibility to ADR nephropathy among C57BL/6 substrains[J]. Exp Anim,2023,72(4):520-525. doi: 10.1538/expanim.23-0003
[12]	Bryant C, Cianciolo R, Govindarajan R, et al. Adriamycin-induced nephropathy is robust in N and modest in J substrain of C57BL/6[J]. Front Cell Dev Biol,2022,10:924751. doi: 10.3389/fcell.2022.924751
[13]	Arif E, Solanki AK, Nihalani D. Adriamycin susceptibility among C57BL/6 substrains[J]. Kidney Int,2016,89(3):721-723. doi: 10.1016/j.kint.2015.10.019
[14]	Pippin JW, Brinkkoetter PT, Cormack-Aboud FC, et al. Inducible rodent models of acquired podocyte diseases[J]. Am J Physiol Renal Physiol,2009,296(2):F213-F229. doi: 10.1152/ajprenal.90421.2008
[15]	Qiu Y, Lei C, Zeng J, et al. Asparagine endopeptidase protects podocytes in adriamycin-induced nephropathy by regulating actin dynamics through cleaving transgelin[J]. Mol Ther,2023,31(11):3337-3354. doi: 10.1016/j.ymthe.2023.09.003
[16]	Wu H, Liu Y, Jia Z, et al. Inhibition of RAC attenuates adriamycin-induced podocyte injury[J]. Biochem Biophys Res Commun,2024,709:149807. doi: 10.1016/j.bbrc.2024.149807
[17]	Shao G, Xu J, Hu C, et al. Podocyte YAP ablation decreases podocyte adhesion and exacerbates FSGS progression through α3β1 integrin[J]. J Pathol,2025,265(1):84-98. doi: 10.1002/path.6370
[18]	Ni Y, Wang X, Yin X, et al. Plectin protects podocytes from adriamycin-induced apoptosis and F-actin cytoskeletal disruption through the integrin α6β4/FAK/p38 MAPK pathway[J]. J Cell Mol Med,2018,22(11):5450-5467.
[19]	Liu L, Li Q, Zhang G. Systemic inflammation accelerates the development of focal segmental glomerulosclerosis in a mouse model of adriamycin induced nephrosis[J]. Sci Rep,2025,15(1):14304. doi: 10.1038/s41598-025-96125-0
[20]	Cui X, Fu J, Luan J, et al. CircZNF609 is involved in the pathogenesis of focal segmental glomerulosclerosis by sponging miR-615-5p[J]. Biochem Biophys Res Commun,2020,531(3):341-349. doi: 10.1016/j.bbrc.2020.07.066
[21]	Ji B, Liu J, Yin Y, et al. Minnelide combined with anti-ANGPTL3-FLD monoclonal antibody completely protects mice with adriamycin nephropathy by promoting autophagy and inhibiting apoptosis[J]. Cell Death Dis,2023,14(9):601. doi: 10.1038/s41419-023-06124-0
[22]	Zhuang Q, Li F, Liu J, et al. Nuclear exclusion of YAP exacerbates podocyte apoptosis and disease progression in adriamycin-induced focal segmental glomerulosclerosis[J]. Lab Invest,2021,101(2):258-270. doi: 10.1038/s41374-020-00503-3
[23]	Barutta F, Kimura S, Hase K, et al. Protective role of the M-Sec-tunneling nanotube system in podocytes[J]. J Am Soc Nephrol,2021,32(5):1114-1130. doi: 10.1681/ASN.2020071076
[24]	Mekada K, Abe K, Murakami A, et al. Genetic differences among C57BL/6 substrains[J]. Exp Anim,2009,58(2):141-149. doi: 10.1538/expanim.58.141
[25]	Simon MM, Greenaway S, White JK, et al. A comparative phenotypic and genomic analysis of C57BL/6J and C57BL/6N mouse strains[J]. Genome Biol,2013,14(7):R82. doi: 10.1186/gb-2013-14-7-r82
[26]	Freeman HC, Hugill A, Dear NT, et al. Deletion of nicotinamide nucleotide transhydrogenase: a new quantitative trait locus accounting for glucose intolerance in C57BL/6J mice[J]. Diabetes,2006,55(7):2153-2156.
[27]	Ronchi JA, Figueira TR, Ravagnani FG, et al. A spontaneous mutation in the nicotinamide nucleotide transhydrogenase gene of C57BL/6J mice results in mitochondrial redox abnormalities[J]. Free Radic Biol Med,2013,63:446-456. doi: 10.1016/j.freeradbiomed.2013.05.049
[28]	Williams JL, Hall CL, Meimaridou E, et al. Loss of Nnt increases expression of oxidative phosphorylation complexes in C57BL/6J hearts[J]. Int J Mol Sci,2021,22(11):6101. doi: 10.3390/ijms22116101
[29]	Williams JL, Paudyal A, Awad S, et al. Mylk3 null C57BL/6N mice develop cardiomyopathy, whereas Nnt null C57BL/6J mice do not[J]. Life Sci Alliance,2020,3(4):e201900593. doi: 10.26508/lsa.201900593
[30]	Ma Q, Grigorescu M, Schreiber A, et al. Genetic background but not intestinal microbiota after co-housing determines hyperoxaluria-related nephrocalcinosis in common inbred mouse strains[J]. Front Immunol,2021,12:673423. doi: 10.3389/fimmu.2021.673423
[31]	Bufi R, Korstanje R. The impact of genetic background on mouse models of kidney disease[J]. Kidney Int,2022,102(1):38-44. doi: 10.1016/j.kint.2022.03.020
[32]	叶秋萍, 陈思慧, 熊云峰, 等. C57BL/6不同亚型对阿霉素肾毒性的易感性[J]. 中华高血压杂志,2024,32(10):941-947. doi: 10.16439/j.issn.1673-7245.2024.10.008
	Ye QP, Chen SH, Xiong YF, et al. The susceptibility of C57BL/6 substrains to adriamycin nephropathy[J]. Chin J Hypertens,2024,32(10):941-947. doi: 10.16439/j.issn.1673-7245.2024.10.008
[33]	Simons M, Hartleben B, Huber TB. Podocyte polarity signaling[J]. Curr Opin Nephrol Hypertens,2009,18(4):324-330.
[34]	Jeansson M, Björck K, Tenstad O, et al. Adriamycin alters glomerular endothelium to induce proteinuria[J]. J Am Soc Nephrol,2009,20(1):114-122. doi: 10.1681/asn.2007111205
[35]	Dai R, Liu H, Han X, et al. Angiopoietin-like-3 knockout protects against glomerulosclerosis in murine adriamycin-induced nephropathy by attenuating podocyte loss[J]. BMC Nephrol,2019,20(1):185. doi: 10.1186/s12882-019-1383-1
[36]	Ji B, Liu J, Ma Y, et al. Minnelide combined with ANGPTL3 knockout completely protects mice with adriamycin nephropathy via suppression of TGF-β1-Smad2 and p53 pathways[J]. Int Immunopharmacol,2023,115:109656. doi: 10.1016/j.intimp.2022.109656

小鼠	组别	尿蛋白/肌酐（mg/g）				F_时间/P	F_交互/P	F_组间/P
小鼠	组别	0周	2周	4周	6周	F_时间/P	F_交互/P	F_组间/P
C57BL/6J	J0	19.63±4.73	24.53±3.80	25.43±2.91	29.94±5.53	781.60/ P<0.0001	267.90/ P<0.0001	759.60/ P<0.0001
	J1	23.70±2.92	89.88±4.64	211.53±12.77	440.66±13.86
	J2	23.32±2.48	90.05±4.90	423.95±35.00	895.40±72.83
C57BL/6N	N0	11.21±2.13	11.43±0.74	12.53±1.83	11.49±1.36	637.10/ P<0.0001	166.10/ P<0.0001	553.10/ P<0.0001
	N1	10.94±1.75	230.84±22.48	917.65±66.94	1758.64±222.61
	N2	10.27±1.34	234.44±6.30	1267.20±70.85	2149.02±139.38

足细胞相关标志蛋白（相对表达量）	J0	J1	J2	N0	N1	N2	F	P
与J0相比，1）：P<0.05; 与J1相比，2）：P<0.05；与N0相比，3）：P<0.05；与N1相比，4）：P<0.05。
肾病蛋白	0.99±0.02	0.38±0.01¹⁾	0.13±0.01¹⁾²⁾	1.21±0.04	0.67±0.03³⁾	0.28±0.03³⁾⁴⁾	712.0	<0.0001
足萼蛋白	1.26±0.14	0.85±0.16¹⁾	0.60±0.05¹⁾	1.21±0.10	0.77±0.13³⁾	0.42±0.08³⁾⁴⁾	25.05	<0.0001
微管蛋白	0.98±0.04	0.69±0.02¹⁾	0.44±0.17¹⁾	1.11±0.13	0.73±0.06³⁾	0.42±0.02³⁾⁴⁾	27.45	<0.0001
足蛋白	1.04±0.16	0.73±0.08¹⁾	0.58±0.15¹⁾	1.16±0.11	0.75±0.06³⁾	0.44±0.13³⁾⁴⁾	15.99	<0.0001
突触足蛋白	1.26±0.14	0.85±0.16	0.60±0.05¹⁾	1.21±0.10	0.77±0.13³⁾	0.42±0.08³⁾⁴⁾	10.34	<0.001
WT1	0.98±0.05	0.75±0.28	0.38±0.19¹⁾²⁾	1.12±0.10	0.68±0.06³⁾	0.32±0.01³⁾⁴⁾	14.60	<0.0001

C57BL/6亚品系在多柔比星诱导FSGS模型中的差异研究

Comparison of doxorubicin-induced FSGS models across C57BL/6 substrains

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