Effect and mechanism of reactive oxygen species/p38 mitogen-activated protein kinase cascade on renal stone formation in rats

XIE Ya-Bin; WANG Fei

doi:10.16098/j.issn.0529-1356.2024.05.012

PDF(4045 KB)

Acta Anatomica Sinica ›› 2024, Vol. 55 ›› Issue (5) : 604-611. DOI: 10.16098/j.issn.0529-1356.2024.05.012

Histology,Embryology and Developmental Biology

Effect and mechanism of reactive oxygen species/p38 mitogen-activated protein kinase cascade on renal stone formation in rats

XIE Ya-bin WANG Fei* WANG Kang-yang LIN Shi-shuai

Author information +

History +

Abstract

Objective To study the effect of reactive oxygen species (ROS)/p38 MAPK cascade reaction on the formation of kidney stones (KS) in rats and explore the mechanism. Methods Fifty SD rats were randomly divided into control group (normal feeding), N-acetylcysteine (NAC) group (intraperitoneally injected 200 mg/kg NAC), KS group (constructed calcium oxalate KS model), KS+NAC group (constructed calcium oxalate KS model, intraperitoneally injected 200 mg/kg NAC), KS+NAC+tunicamycin (TM) group(constructed calcium oxalate KS model, intraperitoneally injected 200 mg/kg NAC and 1 mg/kg TM), with 10 rats in each group. After 4 weeks of administration, 24 hours urine volume and oxalic acid (Ox) of each group were measured, serum creatinine (Cr), urea nitrogen (BUN) and uric acid (UA) levels were detected by automatic biochemical analyzer. HE staining and Von Kossa staining were used to observe the histopathological changes and crystal deposition of the kidney. TUNEL staining was used to detect apoptosis of renal tissue cells. The activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in renal tissue were measured by the kit. DHE fluorescent probes detected the levels of reactive oxygen species (ROS) in kidney tissue. Immunohistochemical staining was used to detect the expressions of microtubule-associated protein 1 light chain 3 B (LC3B) and glucose regulatory protein 78 (GRP78) in renal tissue, and the protein expression of LC3Ⅱ/LC3Ⅰ, Beclin1, GRP78, CCAAT/enhancer binding protein homologous protein (CHOP) in renal tissue was determined by Western blotting. Results Compared with the KS group, Ox in KS+NAC group decreased (P<0.05), BUN, Cr and UA levels decreased (P<0.05), renal tubule dilatation and calcium oxalate crystallization decreased, TUNEL positive cell rate decreased (P<0.05), SOD activity increased and MDA content decreased (P<0.05), ROS levels decreased (P<0.05), LC3B and GRP78 positive staining levels decreased (P<0.05), the relative protein expressions of p-p38 MAPK/p38 MAPK, LC3Ⅱ/LC3Ⅰ, Beclin1, GRP78 and CHOP were down-regulated(P<0.05). Compared with the KS+NAC group, Ox in KS+NAC+TM group increased (P<0.05), BUN, Cr and UA levels also increased (P<0.05), renal tubule dilated significantly, calcium oxalate crystals increased, TUNEL positive cell rate increased (P<0.05), SOD activity decreased and MDA content increased (P<0.05), ROS levels increased (P<0.05), LC3B and GRP78 positive staining levels increased (P<0.05), the relative protein expressions of p-p38 MAPK/p38 MAPK, LC3Ⅱ/LC3Ⅰ, Beclin1, GRP78 and CHOP were also up-regulated (P<0.05). Conclusion ROS/p38 MAPK cascade is involved in promoting KS formation in rats, which is related to the activation of endoplasmic reticulum stress-mediated autophagy pathway.

Key words

/ "> Kidney stone｜Reactive oxygen specie｜p38 mitogen activated protein kinase｜Endoplasmic reticulum stress｜Autophagy｜Western blotting｜Rat

Cite this article

EndNote

Ris (Procite)

Bibtex

Download Citations

XIE Ya-bin WANG Fei WANG Kang-yang LIN Shi-shuai. Effect and mechanism of reactive oxygen species/p38 mitogen-activated protein kinase cascade on renal stone formation in rats[J]. Acta Anatomica Sinica. 2024, 55(5): 604-611 https://doi.org/10.16098/j.issn.0529-1356.2024.05.012

References

［1］ Pavlakou P, Dounousi E, Roumeliotis S, et al. Oxidative stress and the kidney in the space environment［J］. Int J Mol Sci, 2018, 19(10):3176.

［2］ Tormos AM, Taléns-Visconti R, Nebreda AR, et al. p38 MAPK: a dual role in hepatocyte proliferation through reactive oxygen species［J］. Free Radic Res, 2013, 47(11):905-916.

［3］ Zhang ZhY, Zuo QJ, Song X, et al. Study on the effect and mechanism of p38 MAPK pathway mediated by pulmonium extract on calcium oxalate stone formation in rats ［J］. Journal of Sichuan of Traditional Chinese Medicine,2022,40(10):55-58. (in Chinese)

张智源,左庆军,宋旭,等.金钱草提取物介导P38MAPK通路对大鼠草酸钙结石形成的干预效果及机制研究［J］.四川中医,2022,40(10):55-58.

［4］ Liu Y, Chen S, Liu J, et al. Telmisartan inhibits oxalate and calcium oxalate crystal-induced epithelial-mesenchymal transformation via PPAR-γ-AKT/STAT3/p38 MAPK-Snail pathway ［J］. Life Sci, 2020, 241:117108.

［5］ Ge MY，Zhai XY. Study on effect and mechanism of Miniao Paishi Mixture on formation of calcium oxalate kidney stones in rats［J］. Academic Journal of Shanghai University of Traditional Chinese Medicine,2022,36(S1):187-191. (in Chinese)

葛旻垚,翟新宇.泌尿排石合剂对大鼠草酸钙肾结石形成的影响及机制研究［J］.上海中医药大学学报,2022,36(S1):187-191.

［6］ Liang XF, Lu XG, Chen D, et al. Research progress of calcium oxalate stone formation induced by renal tubular epithelial cell injury ［J/OL］. Chinese Journal of Endourology (Electronic Edition),2018,12(4):281-283. (in Chinese)

梁雄发,卢小刚,陈东,等.肾小管上皮细胞损伤促进草酸钙结石形成的研究进展［J/OL］.中华腔镜泌尿外科杂志(电子版),2018,12(4):281-283.

［7］ Wu Y, Zhang J, Li C, et al. The activation of ROS/NF-κB/MMP-9 pathway promotes calcium-induced kidney crystal deposition［J］. Oxid Med Cell Longev, 2021, 2021:8836355.

［8］ Liu Y, Sun Y, Kang J, et al. Role of ROS-induced NLRP3 inflammasome activation in the formation of calcium oxalate nephrolithiasis［J］. Front Immunol, 2022, 13:818625.

［9］ Albert A, Tiwari V, Paul E, et al. Expression of heterologous oxalate decarboxylase in HEK293 cells confers protection against oxalate induced oxidative stress as a therapeutic approach for calcium oxalate stone disease［J］. J Enzyme Inhib Med Chem, 2017, 32(1):426-433.

［10］ Halasi M, Wang M, Chavan TS, et al. ROS inhibitor N-acetyl-L-cysteine antagonizes the activity of proteasome inhibitors［J］. Biochem J, 2013, 454(2):201-208.

［11］ Liu J, Liu Q, Han J, et al. N-Acetylcysteine inhibits patulin-induced apoptosis by affecting ROS-mediated oxidative damage pathway［J］. Toxins (Basel), 2021, 13(9):595.

［12］ Xiong W, Chen H, Lu J, et al. IL-39 increases ROS production and promotes the phosphorylation of p38 MAPK in the apoptotic cardiomyocytes［J］. Folia Histochem Cytobiol, 2021, 59(3):195-202.

［13］ Li C, Chai X, Pan J, et al. β-Hydroxybutyrate alleviates low glucose-induced apoptosis via modulation of ROS-mediated p38 MAPK signaling［J］. J Mol Neurosci, 2022, 72(5):923-938.

［14］ Sharma M, Naura AS, Singla SK. A deleterious interplay between endoplasmic reticulum stress and its functional linkage to mitochondria in nephrolithiasis［J］. Free Radic Biol Med, 2021, 168:70-80.

［15］ Kang J, Sun Y, Deng Y, et al. Autophagy-endoplasmic reticulum stress inhibition mechanism of superoxide dismutase in the formation of calcium oxalate kidney stones［J］. Biomed Pharmacother, 2020, 121:109649.

［16］ Li H, Zhou Y, Xu W, et al. The role of autophagy in calcium oxalate kidney stone: a systematic review of the literature［J］. Front Physiol, 2022, 13:1008264.