Oxoglutarate dehydrogenase-L regulates mitochondrial metabolism reprogramming of renal tubular epithelial cells to alleviate renal fibrosis

YANG Yingxian; HUANG Yinghui; ZHAO Jinghong

doi:10.16016/j.2097-0927.202501053

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Oxoglutarate dehydrogenase-L regulates mitochondrial metabolism reprogramming of renal tubular epithelial cells to alleviate renal fibrosis

Basic Medicine | 更新时间：2025-06-11

- Oxoglutarate dehydrogenase-L regulates mitochondrial metabolism reprogramming of renal tubular epithelial cells to alleviate renal fibrosis
- Journal of Army Medical University Vol. 47, Issue 11, Pages: 1145-1154(2025)
- 作者机构：
  
  陆军军医大学（第三军医大学）第二附属医院肾内科，重庆市肾脏病防治重点实验室，重庆市肾病与泌尿系统疾病临床医学研究中心，重庆
- 作者简介：
- 基金信息：
  
  the Key Program of National Natural Science Foundation of China(82030023);the Natural Science Foundation for Outstanding Young Scholars of China(82322012);the Key Project of Joint Regional Innovation and Development Fund of National Natural Science Foundation of China(U22A20279);the Chongqing Talent Program(CSTC2021YCJH-BGZXM0145;CQYC20220511193)
- DOI：10.16016/j.2097-0927.202501053
  CLC： R692.9;R966;R977.3
- Received：02 February 2025，
  
  Revised：08 April 2025，
  
  Published：15 June 2025
- 稿件说明：
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杨应现，黄英辉，赵景宏.氧戊二酸脱氢酶L调控肾小管上皮细胞线粒体代谢重编程缓解肾纤维化［J］.陆军军医大学学报，2025，47（11）：1145-1154.DOI：10.16016/j.2097-0927.202501053.

ANG Yingxian，HUANG Yinghui，ZHAO Jinghong.Oxoglutarate dehydrogenase-L regulates mitochondrial metabolism reprogramming of renal tubular epithelial cells to alleviate renal fibrosis［J］.J Army Med Univ，2025，47（11）：1145-1154.DOI：10.16016/j.2097-0927.202501053.
杨应现，黄英辉，赵景宏.氧戊二酸脱氢酶L调控肾小管上皮细胞线粒体代谢重编程缓解肾纤维化［J］.陆军军医大学学报，2025，47（11）：1145-1154.DOI：10.16016/j.2097-0927.202501053. DOI：

ANG Yingxian，HUANG Yinghui，ZHAO Jinghong.Oxoglutarate dehydrogenase-L regulates mitochondrial metabolism reprogramming of renal tubular epithelial cells to alleviate renal fibrosis［J］.J Army Med Univ，2025，47（11）：1145-1154.DOI：10.16016/j.2097-0927.202501053. DOI：

摘要

目的

探究氧戊二酸脱氢酶L（oxoglutarate dehydrogenase-L，OGDHL）在肾纤维化中的作用及其机制。

方法

采用随机数字表法将12只雄性野生型C57BL/6J小鼠（8周龄，体质量约25 g）分为对照组与单侧输尿管梗阻（unilateral ureteral obstruction，UUO）组（

=6），通过UUO建立小鼠肾纤维化模型。采用苏木精-伊红（hematoxylin and eosin，HE）染色、免疫荧光染色、免疫组织化学染色、Masson染色检测模型的纤维化程度；培养人近端小管上皮细胞系-2（human kidney-2 proximal tubule cell line，HK-2），分为对照质粒空载体组（Vector组）、过表达OGDHL组（OGDHL OE组）、TGF-β1+对照质粒空载体组（TGF-β1+Vector组）、TGF-β1+过表达OGDHL组（TGF-β1+OGDHL OE组）4组，通过透射电镜观察线粒体形态；通过RT-qPCR、Western blot、免疫荧光染色检测OGDHL及线粒体代谢相关基因的表达水平，并通过试剂盒检测三磷酸腺苷（adenosine triphosphate，ATP）水平，通过安捷伦Seahorse细胞代谢分析仪检测氧耗率（oxygen consumption rate，OCR）、细胞外酸化率（extracellular acidification rate，ECAR）等线粒体代谢相关指标。

结果

UUO术后肾脏肾皮质明显变薄、肾积水显著。HE染色显示肾小管萎缩、管腔扩大；免疫荧光、免疫组织化学染色、Masson染色显示纤维化明显。肾纤维化时，肾脏OGDHL的表达在mRNA水平下降82.9%（

0.001），在蛋白水平下降81.9%（

0.001）。线粒体ATP生成减少（0.970±0.151）µmol/L（

0.01），线粒体结构损伤，最大呼吸能力降低（25.260±1.920） pmol/min（

0.001），脂肪酸氧化相关基因下降50.2%（

0.05），糖酵解增强，ECAR上升，糖酵解相关基因上升为对照组的2.5倍（

0.05）。 OGDHL过表达后线粒体形态学损伤显著改善，ATP生成增加（0.980±0.090） µmol/L（

0.001），能量代谢障碍显著缓解，肾纤维化明显减轻（

0.05）。

结论

OGDHL调控线粒体代谢重编程从而缓解肾小管上皮细胞的纤维化，提示OGDHL可能成为肾纤维化治疗的新靶点。

Abstract

Objective‍　‍To investigate the role and underlying mechanisms of oxoglutarate dehydrogenase-L (OGDHL) in renal fibrosis. Methods‍　‍Twelve male wild-type C57BL/6J mice (8 weeks old

weighting about 25 g) were randomly divided into control group and model group (

=6). A mouse model of renal fibrosis was established by unilateral ureteral obstruction (UUO). HE staining

immunofluorescence assay

and immunohistochemical and Masson staining were applied to assess the extent of ren

al fibrosis. Human kidney-2 proximal tubule cell line (HK-2) were randomly divided into blank plasmid group (Vector)

OGDHL overexpression group (OGDHL OE)

blank plasmid + TGF-β1 treatment group (TGF-β1+Vector)

TGF-β1 treatment and OGDHL overexpression group (TGF-β1+OGDHL OE). Transmission electron microscopy (TEM) was employed to observe mitochondrial morphology. RT-qPCR

Western blotting and immunofluorescence assay were used to detect the expression levels of OGDHL and mitochondrial metabolism-related genes. Adenosine triphosphate (ATP) level

oxygen consumption rate (OCR)

extracellular acidification rate (ECAR) and other mitochondrial metabolism-related indicators were detected with corresponding reagent kits and Agilent Seahorse. Results‍　‍UUO operation resulted in obvious thinning of the renal cortex

significant hydronephrosis. HE staining revealed that tubular atrophy accompanied by dilated tubular lumens in the kidneys. Immunofluorescence

immunohistochemical and Masson staining showed obvious renal fibrosis after UUO operation. In the mouse model of renal fibrosis

the expression of OGDHL was decreased by 82.9% at mRNA level (

0.001) and 81.9% at protein level (

0.001)

the production of mitochondrial ATP was reduced by 0.970±0.151 µmol/L (

0.01)

and notable damaged mitochondrial structure was observed

with maximum breathing capacity decreased by 25.260±1.920 pmol/min (

0.001)

expression levels of fatty acid oxidation-related genes were declined by 50.2% (

0.05)

while glycolysis was enhanced

with the genes related to glycolysis increased by 2.5 times when compared with those in the control group (

0.05). After OGDHL overexpression

mitochondrial morphological damage was significantly improved

ATP production was increased by 0.980±0.090 µmol/L (

0.05)

with significantly alleviated energy metabolism disorder and renal fibrosis (

0.05). Conclusion‍　‍OGDHL regulates mitochondrial metabolic reprogramming of renal tubular epithelial cells to alleviate fibrosis

suggesting that OGDHL may be a new therapeutic target for renal fibrosis.

关键词

Keywords

references

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