---
title: "NSUN4-mediated m5C promotes mitochondrial fission and worsens diabetic nephropathy via the SMURF1/"
id: "pubmed-42661453"
canonical_url: "https://medichelpline.com/clinical-feed/pubmed-42661453"
content_type: "clinical_feed_article"
specialty: "Pharmacology"
source_name: "PubMed / NCBI"
source_url: "https://pubmed.ncbi.nlm.nih.gov/42661453/"
doi: "10.1002/ctm2.70783"
published_at: "2026-09-01T00:00:00.000Z"
evidence_level: "Journal Article"
license: "CC-BY-NC-4.0 / Informational Use"
---
# NSUN4-mediated m5C promotes mitochondrial fission and worsens diabetic nephropathy via the SMURF1/
## Provenance & Clinical Metadata
- **Canonical URL:** https://medichelpline.com/clinical-feed/pubmed-42661453
- **Specialty:** [Pharmacology](https://medichelpline.com/clinical-feed/pharmacology.md)
- **Primary Source:** PubMed / NCBI
- **Source URL:** [Original Journal Publication](https://pubmed.ncbi.nlm.nih.gov/42661453/)
- **DOI:** [10.1002/ctm2.70783](https://doi.org/10.1002%2Fctm2.70783)
- **Published At:** 2026-09-01T00:00:00.000Z
- **Evidence Rating:** Journal Article
## Executive GIST (TL;DR)
- NSUN4, a member of the NOP2/Sun RNA methyltransferase family, is upregulated in diabetic nephropathy (DN) and increases global **m5C** RNA modification levels in patient samples, STZ-induced DN mice, and high‑glucose (HG) HK-2 cells. - Genetic deletion or knockdown of **NSUN4** reduces m5C, alleviates STZ-induced renal injury in mice, and protects HK-2 cells from HG-induced apoptosis. - NSUN4 loss suppresses mitochondrial fission and promotes mitochondrial fusion, shown by changes in mitochondrial morphology, expression of fusion/fission proteins (Mfn1, Mfn2, Fis1, p-Drp1 Ser616, Drp1), mitochondrial membrane potential, ROS, and ATP levels. - NSUN4-mediated m5C enhances **SMURF1** mRNA stability via the m5C reader ALYREF; increased SMURF1 promotes mitochondrial fission and cell apoptosis. - **SMURF1** drives ubiquitination and degradation of **CAMK1**; reduced CAMK1 contributes to mitochondrial fission. Knockdown of CAMK1 reverses the protective effects of SMURF1 depletion on fission and apoptosis. - Rescue experiments show SMURF1 overexpression reverses effects of NSUN4 knockdown, and sh-CAMK1 abolishes effects of sh-SMURF1, supporting a functional NSUN4→SMURF1→CAMK1 axis. - The study integrates in vivo STZ mouse models and in vitro HG HK-2 cell experiments, providing mechanistic evidence that the **NSUN4/SMURF1/CAMK1 axis** promotes mitochondrial fragmentation and accelerates DN progression.
## Clinical Analysis & Structured Key Points
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Affiliations Expand ### Affiliations * 1 Department of Nephrology, The Second Xiangya Hospital of Central South University, Hunan Key Laboratory of Kidney Disease and Blood Purification, Changsha City, China. * 2 National Clinical Research Center for Metabolic Diseases, The Second Xiangya Hospital, Central South University, Changsha City, China. * 3 Department of Nephrology, Hunan Provincial People's Hospital, The First Affiliated Hospital of Hunan Normal University, Changsha City, China. * 4 Department of Nephrology, The First Affiliated Hospital of Changde Vocational Technical College, Changde City, China. * PMID: **42661453** * PMCID: [ PMC13522825 ](https://pmc.ncbi.nlm.nih.gov/articles/PMC13522825/) * DOI: [ 10.1002/ctm2.70783 ](https://doi.org/10.1002/ctm2.70783) Item in Clipboard # NSUN4-dependent m5C modification facilitates mitochondrial fission to accelerate diabetic nephropathy process via regulating SMURF1/CAMK1 axis Danyi Yang et al. Clin Transl Med. 2026 Sep. Show details Display options Display options Format Abstract PubMed PMID Clin Transl Med Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22Clin+Transl+Med%22%5Bjour%5D&sort=date&sort_order=desc) * [ Search in NLM Catalog ](https://www.ncbi.nlm.nih.gov/nlmcatalog?term=%22Clin+Transl+Med%22%5BTitle+Abbreviation%5D) * [ Add to Search ](https://pubmed.ncbi.nlm.nih.gov/42661453/) . 2026 Sep;16(9):e70783. doi: 10.1002/ctm2.70783. ### Authors [Danyi Yang](https://pubmed.ncbi.nlm.nih.gov/?term=Yang+D&cauthor_id=42661453)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42661453/#short-view-affiliation-1 "Department of Nephrology, The Second Xiangya Hospital of Central South University, Hunan Key Laboratory of Kidney Disease and Blood Purification, Changsha City, China.")[ 2 ](https://pubmed.ncbi.nlm.nih.gov/42661453/#short-view-affiliation-2 "National Clinical Research Center for Metabolic Diseases, The Second Xiangya Hospital, Central South University, Changsha City, China."), [Yinyin Chen](https://pubmed.ncbi.nlm.nih.gov/?term=Chen+Y&cauthor_id=42661453)[ 3 ](https://pubmed.ncbi.nlm.nih.gov/42661453/#short-view-affiliation-3 "Department of Nephrology, Hunan Provincial People's Hospital, The First Affiliated Hospital of Hunan Normal University, Changsha City, China."), [Hao Li](https://pubmed.ncbi.nlm.nih.gov/?term=Li+H&cauthor_id=42661453)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42661453/#short-view-affiliation-1 "Department of Nephrology, The Second Xiangya Hospital of Central South University, Hunan Key Laboratory of Kidney Disease and Blood Purification, Changsha City, China."), [Guoyong Liu](https://pubmed.ncbi.nlm.nih.gov/?term=Liu+G&cauthor_id=42661453)[ 4 ](https://pubmed.ncbi.nlm.nih.gov/42661453/#short-view-affiliation-4 "Department of Nephrology, The First Affiliated Hospital of Changde Vocational Technical College, Changde City, China."), [Yang Gao](https://pubmed.ncbi.nlm.nih.gov/?term=Gao+Y&cauthor_id=42661453)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42661453/#short-view-affiliation-1 "Department of Nephrology, The Second Xiangya Hospital of Central South University, Hunan Key Laboratory of Kidney Disease and Blood Purification, Changsha City, China."), [Liyu He](https://pubmed.ncbi.nlm.nih.gov/?term=He+L&cauthor_id=42661453)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42661453/#short-view-affiliation-1 "Department of Nephrology, The Second Xiangya Hospital of Central South University, Hunan Key Laboratory of Kidney Disease and Blood Purification, Changsha City, China.")[ 2 ](https://pubmed.ncbi.nlm.nih.gov/42661453/#short-view-affiliation-2 "National Clinical Research Center for Metabolic Diseases, The Second Xiangya Hospital, Central South University, Changsha City, China.") ### Affiliations * 1 Department of Nephrology, The Second Xiangya Hospital of Central South University, Hunan Key Laboratory of Kidney Disease and Blood Purification, Changsha City, China. * 2 National Clinical Research Center for Metabolic Diseases, The Second Xiangya Hospital, Central South University, Changsha City, China. * 3 Department of Nephrology, Hunan Provincial People's Hospital, The First Affiliated Hospital of Hunan Normal University, Changsha City, China. * 4 Department of Nephrology, The First Affiliated Hospital of Changde Vocational Technical College, Changde City, China. * PMID: **42661453** * PMCID: [ PMC13522825 ](https://pmc.ncbi.nlm.nih.gov/articles/PMC13522825/) * DOI: [ 10.1002/ctm2.70783 ](https://doi.org/10.1002/ctm2.70783) Item in Clipboard Full text links Cite Display options Display options Format Abstract PubMed PMID ## Abstract **Background:** NOP2/Sun RNA methyltransferase family member 4 (NSUN4)-mediated 5-methylcytosine (m5C) modification has been implicated in diabetes-related diseases. However, its role and molecular mechanism in diabetic nephropathy (DN) remain unclear. **Methods:** DN mouse model was constructed by injecting with streptozotocin (STZ). High glucose (HG)-induced HK-2 cells were used to establish a cellular model of DN. The levels of NSUN4, SMAD ubiquitination regulatory factor 1 (SMURF1), calcium/calmodulin-dependent protein kinase 1 (CAMK1), and mitochondrial-related proteins were analyzed using qRT-PCR, western blot, or immunohistochemical staining. Renal injury in mice was evaluated using relevant kits and histological staining. Cell apoptosis, ROS production and proliferation were examined by TUNEL, dihydroethidium, MitoSOX Red and EdU staining. The interaction between SMURF1 and Aly/REF export factor (ALYREF) or CAMK1 was confirmed by RNA immunoprecipitation, Co-IP and ubiquitination assay. **Results:** Upregulation of NSUN4 increased the m5C level in DN. Knockout of NSUN4 alleviated STZ-induced renal injury in mice by repressing renal tubule cell mitochondrial fission and promoting mitochondrial fusion. NSUN4 downregulation reduced mitochondrial fission and promoted mitochondrial fusion to relieve HG-induced HK-2 cell apoptosis. NSUN4-mediated m5C modification promoted SMURF1 mRNA stability by regulating ALYREF. SMURF1 overexpression rescued the suppressive effect of NSUN4 knockdown on cell mitochondrial fission and apoptosis. Besides, SMURF1 facilitated the ubiquitination and degradation of CAMK1. Furthermore, sh-CAMK1 abolished the inhibitory effect of sh-SMURF1 on cell mitochondrial fission and apoptosis. Meanwhile, downregulation of NSUN4 relieved STZ-induced renal injury in DN mice by reducing mitochondrial fission via the SMURF1/CAMK1 axis. **Conclusion:** NSUN4-mediated upregulation of SMURF1 promoted mitochondrial fission to accelerate DN progression via increasing CAMK1 ubiquitination. The discovery of the NSUN4/SMURF1/CAMK1 axis provides new insights into DN pathogenesis. **Keywords:** CAMK1; NSUN4; SMURF1; diabetic nephropathy; m5C modification; mitochondrial. © 2026 The Author(s). Clinical and Translational Medicine published by John Wiley & Sons Australia, Ltd on behalf of Shanghai Institute of Clinical Bioinformatics. [PubMed Disclaimer](https://pubmed.ncbi.nlm.nih.gov/disclaimer/) ## Conflict of interest statement The authors declare no conflicts of interest. ## Figures [ ![FIGURE 1](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/410b/13522825/0e86c3cc3d43/CTM2-16-e70783-g002.gif) ](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/410b/13522825/931045d368ef/CTM2-16-e70783-g002.webp) ** FIGURE 1 ** Effect of NSUN4 on m5C… ** FIGURE 1 ** Effect of NSUN4 on m5C level in DN. (A–F) m5C level was detected… **FIGURE 1** Effect of NSUN4 on m5C level in DN. (A–F) m5C level was detected with LC‐MS/MS and dot blot assay in DN patients (_n_ = 10), STZ‐induced DN mouse models (_n_ = 8), and HG‐induced HK‐2 cells (_n_ = 3). (G–I) The expression levels of 7 m5C writers were tested by qRT‐PCR in DN patients (_n_ = 10), mouse models (_n_ = 8), and cell models (n = 3). (J–L) WB was used to measure NSUN4 protein expression in DN patients (_n_ = 10), mouse models (_n_ = 8), and cell models (_n_ = 3). (M, N) m5C expression was checked using dot blot assay in HG‐induced HK‐2 cells transfected with pc‐NC/pc‐NSUN4/sh‐NC/sh‐NSUN4 (_n_ = 3). The data are presented as the mean ± SD. (A–N) Student's _t_ ‐test. *_p_ < .05, **_p_ < .01, ***_p_ < .001. [ ![FIGURE 2](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/410b/13522825/f08af0c5d207/CTM2-16-e70783-g005.gif) ](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/410b/13522825/8c41fb31ff8c/CTM2-16-e70783-g005.webp) ** FIGURE 2 ** Effect of NSUN4‐cKO on renal… ** FIGURE 2 ** Effect of NSUN4‐cKO on renal injury and mitochondrial fission in STZ‐induced DN mice.… **FIGURE 2** Effect of NSUN4‐cKO on renal injury and mitochondrial fission in STZ‐induced DN mice. (A) The flow chart of the model construction. (B) NSUN4 expression was measured using IHC staining in the kidney tissues of NSUN4‐WT, NSUN4‐cKO, STZ+NSUN4‐WT and STZ+NSUN4‐cKO groups. (C, D) Statistical chart of body weight and blood glucose changes for 12 consecutive weeks. (E–H) Serum BUN, serum Cre, U‐NAG, and ACR levels were detected with related kits. (I–L) Renal tubular damage score and glomerular damage score were assessed to evaluate renal injury by H&E and PAS staining. (M) Cell apoptosis was tested by TUNEL staining. (N) DHE staining was used to assess ROS production. (O) Representative image of fragmented mitochondria observed by TEM. (P) Mfn1, Mfn2, Fis1, p‐Drp1 Ser616, and Drp1 protein levels were evaluated using WB. The data are presented as the mean ± SD. (B, E–H, K–P) One‐way ANOVA with Tukey's post hoc test; (C, D) two‐way ANOVA with Tukey's post hoc test. _n_ = 8, *_p_ < .05, **_p_ < .01, ***_p_ < .001. [ ![FIGURE 3](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/410b/13522825/7fd266799673/CTM2-16-e70783-g009.gif) ](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/410b/13522825/b4fb13d9f6e6/CTM2-16-e70783-g009.webp) ** FIGURE 3 ** Effect of sh‐NSUN4 on HG‐induced… ** FIGURE 3 ** Effect of sh‐NSUN4 on HG‐induced HK‐2 cell apoptosis and mitochondrial fission. (A, B)… **FIGURE 3** Effect of sh‐NSUN4 on HG‐induced HK‐2 cell apoptosis and mitochondrial fission. (A, B) NSUN4 mRNA and protein expression were tested with qRT‐PCR and WB in HK‐2 cells transfected with sh‐NC/sh‐NSUN4. (C–J) HG‐induced HK‐2 cells were transfected with sh‐NC/sh‐NSUN4. (C, D) qRT‐PCR and WB were performed to measure NSUN4 mRNA and protein expression. (E) Mfn1, Mfn2, Fis1, p‐Drp1 Ser616, and Drp1 protein expression was detected using WB. (F) MMP depolarization was checked with JC‐1 staining. (G) Mitochondrial ROS level was examined by MotiSOX Red. (H) ATP level was tested using an ATP assay kit. (I, J) Cell proliferation and apoptosis were examined by EdU assay and TUNEL staining. The data are presented as the mean ± SD. (A, B) Student's _t_ ‐test; (C–J) one‐way ANOVA with Tukey's post hoc test. _n_ = 3, *_p_ < .05, **_p_ < .01, ***_p_ < .001. [ ![FIGURE 4](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/410b/13522825/2902d0c1ff73/CTM2-16-e70783-g010.gif) ](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/410b/13522825/b278e6
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