清肾颗粒通过调控糖酵解重编程及组蛋白 H3K18 乳酸化减轻小鼠肾纤维化

doi:10.12122/j.issn.1673-4254.2026.03.12

南方医科大学学报 ›› 2026, Vol. 46 ›› Issue (3): 582-591.doi: 10.12122/j.issn.1673-4254.2026.03.12

• 基础研究 • 上一篇

清肾颗粒通过调控糖酵解重编程及组蛋白 H3K18 乳酸化减轻小鼠肾纤维化

陈义珍(), 王伟丽, 程梦, 张威, 高怡琳, 洪馨, 张磊, 戴荣(), 王亿平()

安徽中医药大学第一附属医院肾内科，安徽合肥 230031

收稿日期:2025-08-04 出版日期:2026-03-20 发布日期:2026-03-26
通讯作者: 戴荣,王亿平 E-mail:chenyizhen1996@126.com;azydairong@163.com;wypwyp54@aliyun.com
作者简介:陈义珍，博士，医师，E-mail: chenyizhen1996@126.com
基金资助:
国家自然科学基金(82274307);安徽省自然科学基金项目(2308085MH292);安徽省自然科学基金项目(202407290785);安徽省卫生健康科研项目(AHWJ2024BAc20084);安徽省卫生健康科研项目(AHWJ2024Aa30409);安徽中医药大学临床科研项目(2024YFYLCZX28);合肥市科技局生命健康专项(GJ2022SM03);2023年度合肥综合性国家科学中心大健康研究院新安医学与中医药现代化研究所科研项目(2023CXMMTCM018);国家中医优势专科(国中医药医政函[2024]90号)

Qingshen Granules inhibits renal fibrosis in mice by regulating glycolytic reprogramming and H3K18 lactylation

Yizhen CHEN(), Weili WANG, Meng CHENG, Wei ZHANG, Yilin GAO, Xin HONG, Lei ZHANG, Rong DAI(), Yiping WANG()

Department of Nephrology, First Affiliated Hospital of Anhui University of Traditional Chinese Medicine, Hefei 230031, China

Received:2025-08-04 Online:2026-03-20 Published:2026-03-26
Contact: Rong DAI, Yiping WANG E-mail:chenyizhen1996@126.com;azydairong@163.com;wypwyp54@aliyun.com
Supported by:
National Natural Science Foundation of China(82274307)

摘要/Abstract

摘要：

目的探讨清肾颗粒对叶酸诱导的肾纤维化小鼠模型及转化生长因子-β1（TGF-β1）诱导的HK-2细胞的作用及其调控糖酵解重编程、乳酸累积和组蛋白 H3K18 乳酸化的机制。方法构建叶酸诱导的肾纤维化小鼠模型，分为正常组、模型组和清肾颗粒组（4.0 g·kg^-1·d^-1），6只/组，灌胃4周。HE 和 Masson 染色观察肾组织病理；Western blotting检测肾组织纤维化标志（α-SMA、FN、E-cad）、糖酵解关键蛋白（HIF-1α、HK2、PFKM、PKM2、LDHA）及组蛋白乳酸化相关蛋白（Pan Kla、H3/H4 乳酸化位点）表达；免疫荧光检测 α-SMA、FN、E-cad、LDHA、H3K18la表达；ELISA检测血清肌酐（SCR）、尿素氮（BUN）；比色法检测肾组织乳酸含量。细胞实验：构建TGF-β1刺激下的HK-2细胞模型，SD大鼠灌胃制备清肾颗粒含药血清用于细胞实验，分为正常组、模型组和清肾颗粒含药血清组。CCK-8测细胞活力，EdU染色检测增殖；Western blotting检测细胞内 α-SMA、E-cad、LDHA、H3K18la表达；Seahorse XFe96 检测细胞外氧耗率（OCR）和细胞外酸化率（ECAR）；比色法检测细胞内乳酸含量。结果肾脏病理检查提示，与模型组相比，清肾颗粒可以明显减轻小鼠肾组织纤维化（P<0.05）。与模型组相比，清肾颗粒组小鼠肾组织中α-SMA、FN、HIF-1α、HK2、PFKM、PKM2、LDHA、Pan Kla、H3K18la 表达均显著减少，E-cad 表达显著增加（P<0.05）。与模型组相比，清肾颗粒组肾组织内 α-SMA、FN、LDHA 表达显著减少，E-cad 表达显著增加（P<0.05），且LDHA与H3K18la在肾小管的共定位水平降低。此外，与模型组相比，清肾颗粒组小鼠血清 Scr、BUN 水平及肾组织内乳酸含量均显著降低（P<0.05）。细胞实验中，与模型组相比，清肾颗粒含药血清提高细胞活力、促进细胞增殖（P<0.05）。与模型组相比，清肾颗粒含药血清组细胞内α-SMA、LDHA、H3K18la 表达减少（P<0.05）；同时显著提高细胞外OCR、降低ECAR，减少细胞内乳酸含量（P<0.05）。外源性补充乳酸后，与清肾颗粒含药血清组相比，细胞内LDHA、H3K18la及α-SMA的抑制作用被显著逆转（P<0.05）。结论清肾颗粒通过调控糖酵解重编程、减少乳酸堆积及抑制组蛋白H3K18乳酸化，发挥减轻肾纤维化的作用。

关键词: 清肾颗粒, 糖酵解重编程, 组蛋白乳酸化, 肾纤维化

Abstract:

Objective To investigate the effects of Qingshen Granules (QSG) on folic acid (FA)-induced renal fibrosis in mice and TGF-β1-stimulated HK-2 cells and the underlying mechanism. Methods A mouse model of FA-induced renal fibrosis were treated daily with QSG at 4.0 g·kg^-¹ via gavage for 4 weeks. Renal histopathological changes were evaluated using HE and Masson staining, and renal expressions of fibrosis markers, key glycolytic pathway proteins, and histone lactylation-related proteins (Pan Kla, and lactylation sites of H3/H4) were detected using Western blotting. Renal expressions of α-SMA, FN, E-cad, LDHA, and H3K18 lactylation (H3K18la) were also examined with immunofluorescence staining. Serum creatinine (SCR) and blood urea nitrogen (BUN) were measured by ELISA, and renal lactate content was determined with colorimetric assay. In HK-2 cells stimulated with TGF-β1, the effects of QSG-medicated serum from SD rats on cell viability, proliferation, protein expressions of α‑SMA, E-cad, LDHA, and H3K18la, extracellular oxygen consumption rate (OCR), extracellular acidification rate (ECAR), and intracellular lactate levels were analyzed. Results The mouse models of renal fibrosis showed increased renal expressions of α‑SMA, FN, HIF-1α, HK2, PFKM, PKM2, LDHA, Pan Kla, and H3K18la, elevated serum SCR and BUN and renal lactate level, and decreased E-cad expression. QSG treatment effectively reversed these changes and alleviated renal fibrosis in the mouse models. Immunofluorescence staining showed that QSG treatment significantly reversed the elevation of renal α‑SMA, FN and LDHA expressions and reduction of LDHA/H3K18la co-localization, and enhanced E-cad expression. In HK-2 cells, TGF‑β1 treatment significantly reduced cell viability, proliferation and OCR, and increased ECAR, lactate, α‑SMA, LDHA, and H3K18la, which were all reversed by treatment with QSG-medicated serum. Exogenous lactate obviously reversed the inhibitory effects of QSG on these proteins. Conclusion QSG alleviates renal fibrosis in mice by modulating glycolytic reprogramming, reducing lactate accumulation, and inhibiting H3K18la.

Key words: Qingshen Granules, glycolytic reprogra-mming, histone lactylation, renal fibrosis

陈义珍, 王伟丽, 程梦, 张威, 高怡琳, 洪馨, 张磊, 戴荣, 王亿平. 清肾颗粒通过调控糖酵解重编程及组蛋白 H3K18 乳酸化减轻小鼠肾纤维化[J]. 南方医科大学学报, 2026, 46(3): 582-591.

Yizhen CHEN, Weili WANG, Meng CHENG, Wei ZHANG, Yilin GAO, Xin HONG, Lei ZHANG, Rong DAI, Yiping WANG. Qingshen Granules inhibits renal fibrosis in mice by regulating glycolytic reprogramming and H3K18 lactylation[J]. Journal of Southern Medical University, 2026, 46(3): 582-591.

图/表 6

图1 清肾颗粒抑制叶酸诱导的小鼠肾纤维化

Fig.1 QSG inhibits FA-induced renal fibrosis in mice. A, B: HE and Masson staining (Scale bar=50 µm) and the quantitative data (n=6). C, D: Western blotting and quantitative analysis of expression levels of α-SMA, FN and E-cad in the renal tissues in each group (n=3). E, F: Immunofluorescence staining of α-SMA, FN and E-cad (Scale bar=50 µm) and the quantitative data (n=6). G, H: Serum creatinine (SCR) and blood urea nitrogen (BUN) levels of the mice (n=6). *P<0.05 vs Vehicle group; #P<0.05 vs FA group.

图2 清肾颗粒调控叶酸诱导肾纤维化小鼠肾组织的糖酵解重编程

Fig.2 QSG regulates glycolytic reprogramming in renal tissues of the mice with FA-induced renal fibrosis. A, B: Western blotting and quantitative analysis of expression levels of HIF-1α, HK2, PFKM, PKM2 and LDHA in the renal tissues in each group (n=3). C, D: Immunofluorescence analysis of LDHA (Scale bar=50 µm) and the quantitative data (n=6). *P<0.05 vs Vehicle group; #P<0.05 vs FA group.

图3 清肾颗粒抑制小鼠肾组织乳酸堆积及组蛋白乳酸化

Fig.3 QSG inhibits lactate accumulation and histone lactylation in renal tissues of the mice. A: Renal tissue lactate levels in each group (n=6). B: Western blotting of Pan Kla in the renal tissues in each group (n=3). C, D:Western blotting and quantitative analysis of expression levels of H3K9la, H3K14la, H3K18la, H3K23la, H3K56la, H4K5la, H4K8la, H4K12la and H4K16la in the renal tissues in each group (n=3). E: Immunofluorescence co-staining of LDHA (green) and H3K18la (red) in renal tissues (Scale bar=20 µm). F: Quantitative analysis of Pearson colocalization coefficient for LDHA/H3K18la in the renal tissues (n=6). *P<0.05 vs Vehicle group; #P<0.05 vs FA group.

图4 清肾颗粒含药血清抑制 TGF-β1诱导的HK-2细胞纤维化

Fig.4 QSG-medicated serum inhibits TGF-β1-induced fibrosis in HK-2 cells. A: Cell viability of HK-2 cells treated with different concentrations of QSG (n=6). B: Cell viability of TGF‑β1-stimulated HK-2 cells treated with different concentrations of QSG (n=6). C, D: EdU cell proliferation assay (Scale bar=100 μm; n=3). E, F: Western blotting and quantitative analysis of expression levels of α-SMA and E-cad in the renal tissues in each group (n=3). *P<0.05 vs Control group; #P<0.05 vs TGF-β1 group.

图5 清肾颗粒含药血清调控TGF-β1诱导的HK-2细胞糖酵解重编程

Fig.5 QSG-medicated serum regulates TGF-β1-induced glycolytic reprogramming in HK-2 cells. A, B: Changes in extracellular oxygen consumption rate (OCR) in HK-2 cells in each group (n=3). C, D: Changes in extracellular acidification rate (ECAR) in HK-2 cells in each group (n=3). *P<0.05 vs Control group; #P<0.05 vs TGF-β1 group.

图6 清肾颗粒含药血清抑制TGF-β1诱导的HK-2细胞乳酸堆积及组蛋白乳酸化

Fig.6 QSG-medicated serum inhibits TGF-β1-induced lactate accumulation and histone lactylation in HK-2 cells. A: Renal tissue lactate levels in each group (n=6). B, C: Western blotting and quantitative analysis of expression levels of LDHA, H3K18la and α‑SMA in the HK-2 cell in each group (n=3). *P<0.05 vs Control group; #P<0.05 vs TGF‑β1 group; †P<0.05 vs QSG-medicated serum group.

参考文献 33

[1]	Stevens PE, Ahmed SB, Carrero JJ, et al. KDIGO 2024 clinical practice guideline for the evaluation and management of chronic kidney disease[J]. Kidney Int, 2024, 105(4): S117-314. doi：10.1016/j.kint.2023.10.018
[2]	Wang LM, Xu X, Zhang M, et al. Prevalence of chronic kidney disease in China: results from the sixth China chronic disease and risk factor surveillance[J]. JAMA Intern Med, 2023, 183(4): 298-310. doi：10.1001/jamainternmed.2022.6817
[3]	Huang RS, Fu P, Ma L. Kidney fibrosis: from mechanisms to therapeutic medicines[J]. Signal Transduct Target Ther, 2023, 8(1): 129. doi：10.1038/s41392-023-01379-7
[4]	Kang HM, Ahn SH, Choi P, et al. Defective fatty acid oxidation in renal tubular epithelial cells has a key role in kidney fibrosis development[J]. Nat Med, 2015, 21(1): 37-46. doi：10.1038/nm.3762
[5]	Zhu ZJ, Hu JJ, Chen ZW, et al. Transition of acute kidney injury to chronic kidney disease: role of metabolic reprogramming[J]. Metabolism, 2022, 131: 155194. doi：10.1016/j.metabol.2022.155194
[6]	Vander Heiden MG, Cantley LC, Thompson CB. Understanding the Warburg effect: the metabolic requirements of cell proliferation[J]. Science, 2009, 324(5930): 1029-33. doi：10.1126/science.1160809
[7]	Zhang L, Tian MQ, Zhang M, et al. Forkhead box protein K1 promotes chronic kidney disease by driving glycolysis in tubular epithelial cells[J]. Adv Sci (Weinh), 2024, 11(36): e2405325. doi：10.1002/advs.202405325
[8]	Izzo LT, Wellen KE. Histone lactylation links metabolism and gene regulation[J]. Nature, 2019, 574(7779): 492-3. doi：10.1038/d41586-019-03122-1
[9]	Hu XF, Chen W, Yang M, et al. IGFBP5 promotes EndoMT and renal fibrosis through H3K18 lactylation in diabetic nephropathy[J]. Cell Mol Life Sci, 2025, 82(1): 215. doi：10.1007/s00018-025-05718-5
[10]	Wang D, Wang YP, Li CP, et al. Effects of Qingshen Granules on immune function in patients with comorbid chronic renal failure and damp-heat syndrome: a multicenter, randomized, controlled trial[J]. Evid Based Complement Alternat Med, 2020, 2020: 5057894. doi：10.1155/2020/5057894
[11]	Wang YP, Zhang L, Jin H, et al. Based on HIF-1 α/wnt/β-catenin pathway to explore the effect of Qingshen Granules on chronic renal failure patients: a randomized controlled trial[J]. Evid Based Complement Alternat Med, 2019, 2019: 7656105. doi：10.1155/2019/7656105
[12]	Wang YP, Wang D, Jin H, et al. Effects of Qingshen granules on Janus Kinase/signal transducer and activator of transcription signaling pathway in rats with unilateral ureteral obstruction[J]. J Tradit Chin Med, 2018, 38(2): 182-9. doi：10.1016/j.jtcm.2017.04.001
[13]	Zhou WJ, Liang W, Hu MX, et al. Qingshen granules inhibits dendritic cell glycolipid metabolism to alleviate renal fibrosis via PI3K-AKT-mTOR pathway[J]. Phytomedicine, 2024, 135: 156148. doi：10.1016/j.phymed.2024.156148
[14]	戴荣, 曹泽平, 刘传娇, 等. 清肾颗粒通过调控外泌体、miR-330-3p以及CREBBP表达抑制小鼠肾纤维化[J]. 南方医科大学学报, 2024, 44(8): 1431-40. doi：10.12122/j.issn.1673-4254.2024.08.01
[15]	刘敏, 金华, 呼琴, 等. 清肾颗粒含药血清通过miR-23b-5p靶向Nrf2通路减轻NRK-52E细胞转分化[J]. 南方医科大学学报, 2023, 43(12): 2078-85.
[16]	Wang WL, Dai R, Cheng M, et al. Metabolic reprogramming and renal fibrosis: what role might Chinese medicine play[J]? Chin Med, 2024, 19(1): 148. doi：10.1186/s13020-024-01004-x
[17]	Miguel V, Shaw IW, Kramann R. Metabolism at the crossroads of inflammation and fibrosis in chronic kidney disease[J]. Nat Rev Nephrol, 2025, 21(1): 39-56. doi：10.1038/s41581-024-00889-z
[18]	Yang S, Wu H, Li YC, et al. Inhibition of PFKP in renal tubular epithelial cell restrains TGF-β induced glycolysis and renal fibrosis[J]. Cell Death Dis, 2023, 14(12): 816. doi：10.1038/s41419-023-06347-1
[19]	Cao HD, Luo J, Zhang Y, et al. Tuberous sclerosis 1 (Tsc1) mediated mTORC1 activation promotes glycolysis in tubular epithelial cells in kidney fibrosis[J]. Kidney Int, 2020, 98(3): 686-98. doi：10.1016/j.kint.2020.03.035
[20]	Wang ML, Zeng F, Ning FL, et al. Ceria nanoparticles ameliorate renal fibrosis by modulating the balance between oxidative phosphorylation and aerobic glycolysis[J]. J Nanobiotechnology, 2022, 20(1): 3. doi：10.1186/s12951-021-01122-w
[21]	Zhang XX, Chen JC, Lin RH, et al. Lactate drives epithelial-mesenchymal transition in diabetic kidney disease via the H3K14la/KLF5 pathway[J]. Redox Biol, 2024, 75: 103246. doi：10.1016/j.redox.2024.103246
[22]	Wang YT, Li HY, Jiang SM, et al. The glycolytic enzyme PFKFB3 drives kidney fibrosis through promoting histone lactylation-mediated NF‑κB family activation[J]. Kidney Int, 2024, 106(2): 226-40. doi：10.1016/j.kint.2024.04.016
[23]	Xiang TY, Wang XJ, Huang SJ, et al. Inhibition of PKM2 by shikonin impedes TGF‑β1 expression by repressing histone lactylation to alleviate renal fibrosis[J]. Phytomedicine, 2025, 136: 156324. doi：10.1016/j.phymed.2024.156324
[24]	Liu Y, Zhou R, Guo YF, et al. Muscle-derived small extracellular vesicles induce liver fibrosis during overtraining[J]. Cell Metab, 2025, 37(4): 824-41.e8. doi：10.1016/j.cmet.2024.12.005
[25]	Shen Y, Jiang L, Wen P, et al. Tubule-derived lactate is required for fibroblast activation in acute kidney injury[J]. Am J Physiol Renal Physiol, 2020, 318(3): F689-701. doi：10.1152/ajprenal.00229.2019
[26]	Hou Y, Liu DW, Guo ZS, et al. Lactate and lactylation in AKI-to-CKD: epigenetic regulation and therapeutic opportunities[J]. Cell Prolif, 2025, 58(9): e70034. doi：10.1111/cpr.70034
[27]	Wang PW, Xie DX, Xiao T, et al. H3K18 lactylation promotes the progression of arsenite-related idiopathic pulmonary fibrosis via YTHDF1/m6A/NREP[J]. J Hazard Mater, 2024, 461: 132582. doi：10.1016/j.jhazmat.2023.132582
[28]	Wang NX, Wang WW, Wang XQ, et al. Histone lactylation boosts reparative gene activation post-myocardial infarction[J]. Circ Res, 2022, 131(11): 893-908. doi：10.1161/circresaha.122.320488
[29]	Zhou YQ, Yan JX, Huang H, et al. The m⁶A reader IGF₂BP₂ regulates glycolytic metabolism and mediates histone lactylation to enhance hepatic stellate cell activation and liver fibrosis[J]. Cell Death Dis, 2024, 15(3): 189. doi：10.1038/s41419-024-06509-9
[30]	Li WH, Zhou C, Yu L, et al. Tumor-derived lactate promotes resistance to bevacizumab treatment by facilitating autophagy enhancer protein RUBCNL expression through histone H3 lysine 18 lactylation (H3K18la) in colorectal cancer[J]. Autophagy, 2024, 20(1): 114-30. doi：10.1080/15548627.2023.2249762
[31]	Jin Z, Yun L, Cheng P. Tanshinone I reprograms glycolysis metabolism to regulate histone H3 lysine 18 lactylation (H3K18la) and inhibits cancer cell growth in ovarian cancer[J]. Int J Biol Macromol, 2025, 291: 139072. doi：10.1016/j.ijbiomac.2024.139072
[32]	Qiao J, Tan Y, Liu HC, et al. Histone H3K18 and ezrin lactylation promote renal dysfunction in sepsis-associated acute kidney injury[J]. Adv Sci (Weinh), 2024, 11(28): e2307216. doi：10.1002/advs.202307216
[33]	Ye ZH, Sun YS, Yang SY, et al. Lgals3 promotes calcium oxalate crystal formation and kidney injury through histone lactylation-mediated FGFR4 activation[J]. Adv Sci (Weinh), 2025, 12(12): e2413937. doi：10.1002/advs.202413937

清肾颗粒通过调控糖酵解重编程及组蛋白 H3K18 乳酸化减轻小鼠肾纤维化

Qingshen Granules inhibits renal fibrosis in mice by regulating glycolytic reprogramming and H3K18 lactylation

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

图/表 6

参考文献 33

相关文章 8

编辑推荐

Metrics

本文评价

[1]	呼琴, 金华. 清肾颗粒通过调控miR-23b及Nrf2通路改善慢性肾脏病湿热证患者的肾功能：基于网络药理学和临床试验[J]. 南方医科大学学报, 2025, 45(9): 1867-1879.
[2]	卢晓宇, 刘智慧, 刘烨, 庞天霄, 卞蓉, 郭玲, 何学红. 参芪泄浊饮通过调控Rap1/MAPK/FoxO3a信号通路改善氧化应激及炎症反应延缓大鼠肾纤维化[J]. 南方医科大学学报, 2025, 45(12): 2585-2597.
[3]	戴荣, 曹泽平, 刘传娇, 葛永, 程梦, 王伟丽, 陈义珍, 张磊, 王亿平. 清肾颗粒通过调控外泌体、miR-330-3p以及CREBBP表达抑制小鼠肾纤维化[J]. 南方医科大学学报, 2024, 44(8): 1431-1440.
[4]	刘敏, 金华, 呼琴, 陈诺, 张叶青, 王亿平. 清肾颗粒含药血清通过miR-23b-5p靶向Nrf2通路减轻NRK-52E细胞转分化[J]. 南方医科大学学报, 2023, 43(12): 2078-2085.
[5]	沈炳香, 王法财, 周艺, 李婷, 何春远, 赵为陈. 人参皂苷Rh2调控盘状结构域受体1对糖尿病大鼠肾纤维化和细胞凋亡的影响[J]. 南方医科大学学报, 2021, 41(7): 1107-1113.
[6]	张磊，王亿平，金华，王东，魏玲，任克军，茅燕萍. 清肾颗粒对肾性贫血患者炎症/铁调素轴及铁代谢的干预机制：单中心前瞻性随机对照研究[J]. 南方医科大学学报, 2019, 39(10): 1155-.
[7]	王瑜，王博士，惠旭，乔君，李委振，孙楠. 可诱导共刺激分子介导的Th17细胞极化对自发性高血压大鼠肾纤维化的影响[J]. 南方医科大学学报, 2018, 38(05): 534-.
[8]	谢潮鑫，孟猛，殷先锋，何凤玲，叶汉深，谢栋. 天然虾青素对抗肾纤维化及细胞凋亡的作用[J]. 南方医科大学学报, 2013, 33(02): 305-.