Renal tubular epithelial cell-specific c-kit gene knockout alleviates renal interstitial fibrosis induced by unilateral ischemia-reperfusion injury in mice

doi:10.3877/cma.j.issn.2095-3216.2026.05.002

Abstract

Abstract:

Objective

To investigate the effects of renal tubular epithelial cell-specific c-kit gene knockout on macrophage infiltration and renal interstitial fibrosis following unilateral ischemia-reperfusion injury (uIRI) in mice.

Methods

Eight-week-old male renal tubular epithelial cell-specific c-kit knockout mice (Ggt1-Cre c-kit^-/-), c-kit^loxp/loxp mice, and wild-type (WT) C57BL/6J mice were used to establish the uIRI model. Experimental mice were divided into six groups (n= 12 per group): Ggt1-Cre c-kit^-/- uIRI group, c-kit^loxp/loxp uIRI group, C57BL/6J uIRI group (WT uIRI group), Ggt1-Cre c-kit^-/- sham group, c-kit^loxp/loxp sham group, and C57BL/6J sham group (WT sham group). Kidney tissue and blood samples were collected at 1, 3, 14, and 28 days post-operation. Serum creatinine (SCr) and blood urea nitrogen (BUN) levels were measured, and renal histopathological changes were evaluated. Sirius red staining was used to assess renal interstitial fibrosis. Macrophage infiltration was examined by immunohistochemistry. The protein and mRNA expression levels of α-smooth muscle actin (α-SMA), vimentin, and collagen I were detected by Western blotting and RT-qPCR, respectively. ANOVA software was used for statistical analysis.

Results

Compared with the WT uIRI group and c-kit^loxp/loxp uIRI group, the Ggt1-Cre c-kit^-/- uIRI group exhibited significantly lower SCr and BUN levels at all time points after uIRI operation (all P<0.05). On 3 and 14 days post-operation, the number of infiltrating macrophages in the Ggt1-Cre c-kit^-/- uIRI group was significantly reduced compared with the other two uIRI groups. On postoperative day 28, the renal interstitial fibrosis score and the mRNA and protein expression levels of fibrosis markers (α-SMA, vimentin, and collagen I) in the Ggt1-Cre c-kit^-/- uIRI group were significantly lower than those in the other two uIRI groups (all P<0.05).

Conclusion

Renal tubular epithelial cell-specific c-kit gene knockout significantly attenuates renal interstitial fibrosis and inhibits macrophage infiltration, suggesting that c-kit gene may serve as a new therapeutic target for kidney diseases.

Key words: Renal tubular epithelial cell, c-kit gene, Gene knockout, Ischemia-reperfusion injury, Macrophage, Renal interstitial fibrosis

Zenghui Xing, Wenkai Guo, Chao Liu, Hongfeng Wang, Shengchun Zheng, Yan Chen, Bing Han, Yixuan Zhang, Guixiu Wang, Xiangmei Chen, Xuefeng Sun. Renal tubular epithelial cell-specific c-kit gene knockout alleviates renal interstitial fibrosis induced by unilateral ischemia-reperfusion injury in mice[J]. Chinese Journal of Kidney Disease Investigation(Electronic Edition), 2026, 15(05): 253-259.

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Figures/Tables 7

References 13

[1]	Ferenbach DA, Bonventre JV. Mechanisms of maladaptive repair after AKI leading to accelerated kidney ageing and CKD [J]. Nat Rev Nephrol, 2015, 11(5): 264-276.
[2]	张一绚，韩冰，刘超，等. 年轻化内环境改善老年小鼠肾缺血再灌注损伤诱导的肾间质纤维化[J/OL]. 中华肾病研究电子杂志，2024, 13(3): 129-133.
[3]	Huang Q, Liu D, Cui S, et al. Effect of youthful blood environment and its key stem cell factor on renal interstitial fibrosis in elderly mice [J]. Gerontology, 2023, 69(5): 628-640.
[4]	Stokman G, Stroo I, Claessen N, et al. Stem cell factor expression after renal ischemia promotes tubular epithelial survival [J]. PLoS One, 2010, 5(12): e14386.
[5]	Grgic I, Campanholle G, Bijol V, et al. Targeted proximal tubule injury triggers interstitial fibrosis and glomerulosclerosis [J]. Kidney Int, 2012, 82(2): 172-183.
[6]	Song Y, Li N, Jiang S, et al. Microbiota-derived H₂S induces c-kit^＋ cDC1 autophagic cell death and liver inflammation in metabolic dysfunction-associated steatohepatitis [J]. Nat Commun, 2025, 16(1): 2222.
[7]	Duan JL, Liu JJ, Ruan B, et al. Age-related liver endothelial zonation triggers steatohepatitis by inactivating pericentral endothelium-derived c-kit[J]. Nat Aging, 2023, 3(3): 258-274.
[8]	Song L, Chang X, Hu L, et al. Accelerating wound closure with Metrnl in normal and diabetic mouse skin [J]. Diabetes, 2023, 72(11): 1692-1706.
[9]	Alexopoulos E, Seron D, Hartley RB, et al. The role of interstitial infiltrates in IgA nephropathy: a study with monoclonal antibodies [J]. Nephrol Dial Transplant, 2009, 24(1): 187-195.
[10]	Nilsson G, Butterfield JH, Nilsson K, et al. Stem cell factor is chemotactic for human mast cells [J]. J Immunol, 1994, 153(8): 3718-3723.
[11]	Bradding P, Feather LH, Wilson S, et al. Immunolocalization of cytokines in the nasal mucosa of normal and perennial rhinitic subjects: the mast cell as a source of IL-4, IL-5, and IL-6 in human allergic mucosal inflammation [J]. J Immunol, 1993, 151(7): 3853-3865.
[12]	Zhang YL, Tang TT, Wang B, et al. Identification of a novel ECM remodeling macrophage subset in AKI to CKD transition by integrative spatial and single-cell analysis [J]. Adv Sci (Weinh), 2024, 11(38): e2309752.
[13]	Ma T, Li X, Zhu Y, et al. Excessive activation of notch signaling in macrophages promote kidney inflammation, fibrosis, and necroptosis [J]. Front Immunol, 2022, 13: 835879.

引物组	扩增子长度(bp)	基因型	序列(5′→3′)
c-kitloxp正向引物	188/247	WT/c-kit^loxp	AATGATGTTCATCGAGCATGAGAG
c-kitloxp反向引物			GAGACATGAACATTAAGGGATGCTG
Ggt1-Cre正向引物	428	Ggt1-Cre	CATCACATCAG GCACCCCAGAA
Ggt1-Cre反向引物			GAACATCTTCAGGTTCTGCGGGA

引物组	扩增子长度(bp)	基因型	序列(5′→3′)
c-kitloxp正向引物	188/247	WT/c-kit^loxp	AATGATGTTCATCGAGCATGAGAG
c-kitloxp反向引物			GAGACATGAACATTAAGGGATGCTG
Ggt1-Cre正向引物	428	Ggt1-Cre	CATCACATCAG GCACCCCAGAA
Ggt1-Cre反向引物			GAACATCTTCAGGTTCTGCGGGA

检测指标	种属	上游引物序列(5′-3′)	下游引物序列(5′-3′)
18S	小鼠	CCTCTATGCCAACACAGTGCTGTCT	GCTCAGGAGGAGCAATGATCTTGA
α-平滑肌肌动蛋白	小鼠	AGCCATCTTTCATTGGGATGG	CCCCTGACAGGACGTTGTTA
波形蛋白	小鼠	GGCTGCGAGAGAAATTGCAGG	AAGACGTGCCAGAGAAGCATT
Ⅰ型胶原	小鼠	ACACGCAAGGCAATGAGACT	TGGTCACTTGCACTGGTTGA

检测指标	种属	上游引物序列(5′-3′)	下游引物序列(5′-3′)
18S	小鼠	CCTCTATGCCAACACAGTGCTGTCT	GCTCAGGAGGAGCAATGATCTTGA
α-平滑肌肌动蛋白	小鼠	AGCCATCTTTCATTGGGATGG	CCCCTGACAGGACGTTGTTA
波形蛋白	小鼠	GGCTGCGAGAGAAATTGCAGG	AAGACGTGCCAGAGAAGCATT
Ⅰ型胶原	小鼠	ACACGCAAGGCAATGAGACT	TGGTCACTTGCACTGGTTGA

肾功能指标	分组	假手术	uIRI 1 d	uIRI 3 d	uIRI 14 d	uIRI 28 d
血清肌酐(μmol/L)	WT组	12.301±0.466	179.094±9.407^ab	50.420±8.807^ab	28.335±2.880^ab	20.917±0.798^b
	c-kit^loxp/loxp组	12.949±2.137	175.612±5.704^ac	52.197±4.542^ac	28.073±0.324^ac	20.077±2.196^b
	Ggt1-Cre c-kit^－/－组	12.253±3.031	114.227±9.124^a	35.710±1.040^a	23.189±6.712^a	16.907±1.642
血尿素氮(mmol/L)	WT组	6.923±1.714	51.143±4.493^ab	27.472±2.560^ab	20.672±2.560^ab	12.552±0.756^ab
	c-kit^loxp/loxp组	6.610±1.286	51.326±1.248^ab	27.400±0.544^ab	20.637±0.574^ab	12.947±0.962^ab
	Ggt1-Cre c-kit^－/－组	6.886±0.792	36.162±2.876^a	22.706±0.419^a	17.490±1.325^a	10.482±0.270

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