Inhibition of LTBP2 Suppresses High Glucose-Induced Proliferation, Fibrosis, and Inflammation in Glomerular Mesangial Cells by Disrupting the PI3K/Akt/NF-κB Pathway
Abstract
Latent transforming growth factor-β binding protein-2 (LTBP2) plays a significant role in tissue fibrosis. This research aimed to elucidate whether LTBP2 influences the progression of diabetic nephropathy (DN) through the phosphatidylinositol 3-kinases/protein kinase B (PI3K/Akt)/nuclear factor kappa-B (NF-κB) pathway.
The HBZY-1 cells were exposed to high glucose to create diabetic nephropathy cell model. LTBP2 levels were examined by Western blot and immunofluorescence. After verifying the transfection efficiency of si-LTBP2, cell counting kit-8, 5-ethynyl-2-deoxyuridine staining, Western blot, flow cytometry and immunofluorescence were utilized to assess the proliferation, apoptosis and fibrosis of HBZY-1 cells, respectively. Collagen deposition was also detected by Sirius red staining, and inflammatory factors levels were determined by Elisa. PI3K/Akt/NF-κB pathway activators were applied to explore whether LTBP2 silencing could play a role in DN by modulating this pathway.
After treatment with high glucose, the expression of LTBP2 was elevated in HBZY-1 cells. LTBP2 silencing hindered the aberrant proliferation of HBZY-1 cells, with no significant effect on apoptosis; meanwhile, it reduced fibrosis, decreased collagen content, and decreased inflammatory factors levels in HBZY-1 cells. Following treatment with high glucose, the PI3K, Akt, and p65 phosphorylation levels were increased, whereas silencing LTBP2 reduced them. Activators of the PI3K/Akt/NF-κB pathway weakened the inhibition of LTBP2 silencing on cell proliferation, fibrosis, and inflammation.
In conclusion, silencing of LTBP2 weakened the proliferation, fibrosis, and inflammation of HBZY-1 cells treated with high glucose by hindering the PI3K/Akt/NF-κB pathway. This research offers a new reference for the targeted therapy of DN.
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7. Selby NM, Taal MW. An updated overview of diabetic nephropathy: Diagnosis, prognosis, treatment goals and latest guidelines. Diabetes Obes Metab. 2020;22 Suppl 1:3-15.
8. Kriz W, Löwen J, Gröne HJ. The complex pathology of diabetic nephropathy in humans. Nephrol Dial Transplant. 2023;38(10):2109-19.
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10. Bodmer NK, Knutsen RH, Roth RA, Castile RM, Brodt MD, Gierasch CM, et al. Multi-organ phenotypes in mice lacking latent TGFβ binding protein 2 (LTBP2). Dev Dyn. 2024;253(2):233-54.
11. Liu Z, Wang T, Shi X, Wang X, Ren W, Huang B, et al. Identification of LTBP2 gene polymorphisms and their association with thoracolumbar vertebrae number, body size, and carcass traits in Dezhou donkeys. Frontiers in genetics. 2022;13:969959.
12. Wang M, Wang M, Zhao J, Xu H, Xi Y, Yang H. Dengzhan Shengmai capsule attenuates cardiac fibrosis in post-myocardial infarction rats by regulating LTBP2 and TGF-β1/Smad3 pathway. Phytomedicine. 2023;116:154849.
13. Zou M, Hu X, Song W, Gao H, Wu C, Zheng W, et al. Plasma LTBP2 as a potential biomarker in differential diagnosis of connective tissue disease-associated interstitial lung disease and idiopathic pulmonary fibrosis: a pilot study. Clin Exp Med. 2023;23(8):4809-16.
14. Zhang L, Tan J, Liu Y, Luo M. Curcumin relieves arecoline-induced oral submucous fibrosis via inhibiting the LTBP2/NF-κB axis. Oral Dis. 2024;30(4):2314-24.
15. Zeng L, Li J, Gao F, Song Y, Wei L, Qu N, et al. SGLT2i improves kidney senescence by down-regulating the expression of LTBP2 in SAMP8 mice. J Cell Mol Med. 2024;28(6):e18176.
16. Deng RM, Zhou J. The role of PI3K/AKT signaling pathway in myocardial ischemia-reperfusion injury. Int Immunopharmacol. 2023;123:110714.
17. Deng S, Leong HC, Datta A, Gopal V, Kumar AP, Yap CT. PI3K/AKT Signaling Tips the Balance of Cytoskeletal Forces for Cancer Progression. Cancers (Basel). 2022;14(7).
18. Long HZ, Cheng Y, Zhou ZW, Luo HY, Wen DD, Gao LC. PI3K/AKT Signal Pathway: A Target of Natural Products in the Prevention and Treatment of Alzheimer's Disease and Parkinson's Disease. Front Pharmacol. 2021;12:648636.
19. Li L, Jiang W, Yu B, Liang H, Mao S, Hu X, et al. Quercetin improves cerebral ischemia/reperfusion injury by promoting microglia/macrophages M2 polarization via regulating PI3K/Akt/NF-κB signaling pathway. Biomed Pharmacother. 2023;168:115653.
20. Li D, Guo YY, Cen XF, Qiu HL, Chen S, Zeng XF, et al. Lupeol protects against cardiac hypertrophy via TLR4-PI3K-Akt-NF-κB pathways. Acta Pharmacol Sin. 2022;43(8):1989-2002.
21. Wan F, Peng L, Zhu C, Zhang X, Chen F, Liu T. Knockdown of Latent Transforming Growth Factor-β (TGF-β)-Binding Protein 2 (LTBP2) Inhibits Invasion and Tumorigenesis in Thyroid Carcinoma Cells. Oncol Res. 2017;25(4):503-10.
22. Pang XF, Lin X, Du JJ, Zeng DY. LTBP2 knockdown by siRNA reverses myocardial oxidative stress injury, fibrosis and remodelling during dilated cardiomyopathy. Acta Physiol (Oxf). 2020;228(3):e13377.
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24. Wang J, Niu Y, Luo L, Lu Z, Chen Q, Zhang S, et al. Decoding ceRNA regulatory network in the pulmonary artery of hypoxia-induced pulmonary hypertension (HPH) rat model. Cell Biosci. 2022;12(1):27.
25. Xiang E, Han B, Zhang Q, Rao W, Wang Z, Chang C, et al. Human umbilical cord-derived mesenchymal stem cells prevent the progression of early diabetic nephropathy through inhibiting inflammation and fibrosis. Stem Cell Res Ther. 2020;11(1):336.
26. Li X, Lu L, Hou W, Huang T, Chen X, Qi J, et al. Epigenetics in the pathogenesis of diabetic nephropathy. Acta Biochim Biophys Sin (Shanghai). 2022;54(2):163-72.
27. Kanwar YS, Sun L, Xie P, Liu FY, Chen S. A glimpse of various pathogenetic mechanisms of diabetic nephropathy. Annu Rev Pathol. 2011;6:395-423.
28. Sindhu D, Sharma GS, Kumbala D. Management of diabetic kidney disease: where do we stand?: A narrative review. Medicine (Baltimore). 2023;102(13):e33366.
29. Wu X, Pan C, Chen R, Zhang S, Zhai Y, Guo H. BML-111 attenuates high glucose-induced inflammation, oxidative stress and reduces extracellular matrix accumulation via targeting Nrf2 in rat glomerular mesangial cells. Int Immunopharmacol. 2020;79:106108.
30. Liu H, Chen W, Lu P, Ma Y, Liang X, Liu Y. Ginsenoside Rg1 attenuates the inflammation and oxidative stress induced by diabetic nephropathy through regulating the PI3K/AKT/FOXO3 pathway. Annals of translational medicine. 2021;9(24):1789.
31. Jin Q, Liu T, Qiao Y, Liu D, Yang L, Mao H, et al. Oxidative stress and inflammation in diabetic nephropathy: role of polyphenols. Front Immunol. 2023;14:1185317.
32. Calle P, Hotter G. Macrophage Phenotype and Fibrosis in Diabetic Nephropathy. Int J Mol Sci. 2020;21(8).
33. Liu Y, Xu K, Xiang Y, Ma B, Li H, Li Y, et al. Role of MCP-1 as an inflammatory biomarker in nephropathy. Front Immunol. 2023;14:1303076.
34. Rayego-Mateos S, Morgado-Pascual JL, Opazo-Ríos L, Guerrero-Hue M, García-Caballero C, Vázquez-Carballo C, et al. Pathogenic Pathways and Therapeutic Approaches Targeting Inflammation in Diabetic Nephropathy. Int J Mol Sci. 2020;21(11).
35. Guo W, Song Y, Sun Y, Du H, Cai Y, You Q, et al. Systemic immune-inflammation index is associated with diabetic kidney disease in Type 2 diabetes mellitus patients: Evidence from NHANES 2011-2018. Front Endocrinol (Lausanne). 2022;13:1071465.
36. Theocharis AD, Manou D, Karamanos NK. The extracellular matrix as a multitasking player in disease. FEBS J. 2019;286(15):2830-69.
37. Chaudhuri O, Cooper-White J, Janmey PA, Mooney DJ, Shenoy VB. Effects of extracellular matrix viscoelasticity on cellular behaviour. Nature. 2020;584(7822):535-46.
38. Li HD, You YK, Shao BY, Wu WF, Wang YF, Guo JB, et al. Roles and crosstalks of macrophages in diabetic nephropathy. Front Immunol. 2022;13:1015142.
39. Qin W, Cao L, Massey IY. Role of PI3K/Akt signaling pathway in cardiac fibrosis. Mol Cell Biochem. 2021;476(11):4045-59.
40. Wang J, Hu K, Cai X, Yang B, He Q, Wang J, et al. Targeting PI3K/AKT signaling for treatment of idiopathic pulmonary fibrosis. Acta pharmaceutica Sinica B. 2022;12(1):18-32.
41. Wang T, Wen X, Zhang Z, Xie M, Zhou J. Phillyrin ameliorates diabetic nephropathy through the PI3K/Akt/GSK-3β signalling pathway in streptozotocin-induced diabetic mice. Hum Exp Toxicol. 2021;40(12_suppl):S487-s96.
42. Wang J, Jiang C, Li N, Wang F, Xu Y, Shen Z, et al. The circEPSTI1/mir-942-5p/LTBP2 axis regulates the progression of OSCC in the background of OSF via EMT and the PI3K/Akt/mTOR pathway. Cell Death Dis. 2020;11(8):682.
43. Wu H, Xu F, Huang X, Li X, Yu P, Zhang L, et al. Lupenone improves type 2 diabetic nephropathy by regulating NF-κB pathway-mediated inflammation and TGF-β1/Smad/CTGF-associated fibrosis. Phytomedicine. 2023;118:154959.
44. Zou M, Zou J, Hu X, Zheng W, Zhang M, Cheng Z. Latent Transforming Growth Factor-β Binding Protein-2 Regulates Lung Fibroblast-to-Myofibroblast Differentiation in Pulmonary Fibrosis via NF-κB Signaling. Front Pharmacol. 2021;12:788714.
2023)]. Wien Klin Wochenschr. 2023;135(Suppl 1):7-17.
2. Sun H, Saeedi P, Karuranga S, Pinkepank M, Ogurtsova K, Duncan BB, et al. IDF Diabetes Atlas: Global, regional and country-level diabetes prevalence estimates for 2021 and projections for 2045. Diabetes Res Clin Pract. 2022;183:109119.
3. Thipsawat S. Early detection of diabetic nephropathy in patient with type 2 diabetes mellitus: A review of the literature. Diab Vasc Dis Res. 2021;18(6):14791641211058856.
4. Sagoo MK, Gnudi L. Diabetic Nephropathy: An Overview. Methods Mol Biol. 2020;2067:3-7.
5. Samsu N. Diabetic Nephropathy: Challenges in Pathogenesis, Diagnosis, and Treatment. BioMed Res Int. 2021;2021:1497449.
6. Lu Y, Liu D, Feng Q, Liu Z. Diabetic Nephropathy: Perspective on Extracellular Vesicles. Front Immunol. 2020;11:943.
7. Selby NM, Taal MW. An updated overview of diabetic nephropathy: Diagnosis, prognosis, treatment goals and latest guidelines. Diabetes Obes Metab. 2020;22 Suppl 1:3-15.
8. Kriz W, Löwen J, Gröne HJ. The complex pathology of diabetic nephropathy in humans. Nephrol Dial Transplant. 2023;38(10):2109-19.
9. Ahmad AA, Draves SO, Rosca M. Mitochondria in Diabetic Kidney Disease. Cells. 2021;10(11).
10. Bodmer NK, Knutsen RH, Roth RA, Castile RM, Brodt MD, Gierasch CM, et al. Multi-organ phenotypes in mice lacking latent TGFβ binding protein 2 (LTBP2). Dev Dyn. 2024;253(2):233-54.
11. Liu Z, Wang T, Shi X, Wang X, Ren W, Huang B, et al. Identification of LTBP2 gene polymorphisms and their association with thoracolumbar vertebrae number, body size, and carcass traits in Dezhou donkeys. Frontiers in genetics. 2022;13:969959.
12. Wang M, Wang M, Zhao J, Xu H, Xi Y, Yang H. Dengzhan Shengmai capsule attenuates cardiac fibrosis in post-myocardial infarction rats by regulating LTBP2 and TGF-β1/Smad3 pathway. Phytomedicine. 2023;116:154849.
13. Zou M, Hu X, Song W, Gao H, Wu C, Zheng W, et al. Plasma LTBP2 as a potential biomarker in differential diagnosis of connective tissue disease-associated interstitial lung disease and idiopathic pulmonary fibrosis: a pilot study. Clin Exp Med. 2023;23(8):4809-16.
14. Zhang L, Tan J, Liu Y, Luo M. Curcumin relieves arecoline-induced oral submucous fibrosis via inhibiting the LTBP2/NF-κB axis. Oral Dis. 2024;30(4):2314-24.
15. Zeng L, Li J, Gao F, Song Y, Wei L, Qu N, et al. SGLT2i improves kidney senescence by down-regulating the expression of LTBP2 in SAMP8 mice. J Cell Mol Med. 2024;28(6):e18176.
16. Deng RM, Zhou J. The role of PI3K/AKT signaling pathway in myocardial ischemia-reperfusion injury. Int Immunopharmacol. 2023;123:110714.
17. Deng S, Leong HC, Datta A, Gopal V, Kumar AP, Yap CT. PI3K/AKT Signaling Tips the Balance of Cytoskeletal Forces for Cancer Progression. Cancers (Basel). 2022;14(7).
18. Long HZ, Cheng Y, Zhou ZW, Luo HY, Wen DD, Gao LC. PI3K/AKT Signal Pathway: A Target of Natural Products in the Prevention and Treatment of Alzheimer's Disease and Parkinson's Disease. Front Pharmacol. 2021;12:648636.
19. Li L, Jiang W, Yu B, Liang H, Mao S, Hu X, et al. Quercetin improves cerebral ischemia/reperfusion injury by promoting microglia/macrophages M2 polarization via regulating PI3K/Akt/NF-κB signaling pathway. Biomed Pharmacother. 2023;168:115653.
20. Li D, Guo YY, Cen XF, Qiu HL, Chen S, Zeng XF, et al. Lupeol protects against cardiac hypertrophy via TLR4-PI3K-Akt-NF-κB pathways. Acta Pharmacol Sin. 2022;43(8):1989-2002.
21. Wan F, Peng L, Zhu C, Zhang X, Chen F, Liu T. Knockdown of Latent Transforming Growth Factor-β (TGF-β)-Binding Protein 2 (LTBP2) Inhibits Invasion and Tumorigenesis in Thyroid Carcinoma Cells. Oncol Res. 2017;25(4):503-10.
22. Pang XF, Lin X, Du JJ, Zeng DY. LTBP2 knockdown by siRNA reverses myocardial oxidative stress injury, fibrosis and remodelling during dilated cardiomyopathy. Acta Physiol (Oxf). 2020;228(3):e13377.
23. Dong Z, Sun Y, Wei G, Li S, Zhao Z. Ergosterol Ameliorates Diabetic Nephropathy by Attenuating Mesangial Cell Proliferation and Extracellular Matrix Deposition via the TGF-β1/Smad2 Signaling Pathway. Nutrients. 2019;11(2).
24. Wang J, Niu Y, Luo L, Lu Z, Chen Q, Zhang S, et al. Decoding ceRNA regulatory network in the pulmonary artery of hypoxia-induced pulmonary hypertension (HPH) rat model. Cell Biosci. 2022;12(1):27.
25. Xiang E, Han B, Zhang Q, Rao W, Wang Z, Chang C, et al. Human umbilical cord-derived mesenchymal stem cells prevent the progression of early diabetic nephropathy through inhibiting inflammation and fibrosis. Stem Cell Res Ther. 2020;11(1):336.
26. Li X, Lu L, Hou W, Huang T, Chen X, Qi J, et al. Epigenetics in the pathogenesis of diabetic nephropathy. Acta Biochim Biophys Sin (Shanghai). 2022;54(2):163-72.
27. Kanwar YS, Sun L, Xie P, Liu FY, Chen S. A glimpse of various pathogenetic mechanisms of diabetic nephropathy. Annu Rev Pathol. 2011;6:395-423.
28. Sindhu D, Sharma GS, Kumbala D. Management of diabetic kidney disease: where do we stand?: A narrative review. Medicine (Baltimore). 2023;102(13):e33366.
29. Wu X, Pan C, Chen R, Zhang S, Zhai Y, Guo H. BML-111 attenuates high glucose-induced inflammation, oxidative stress and reduces extracellular matrix accumulation via targeting Nrf2 in rat glomerular mesangial cells. Int Immunopharmacol. 2020;79:106108.
30. Liu H, Chen W, Lu P, Ma Y, Liang X, Liu Y. Ginsenoside Rg1 attenuates the inflammation and oxidative stress induced by diabetic nephropathy through regulating the PI3K/AKT/FOXO3 pathway. Annals of translational medicine. 2021;9(24):1789.
31. Jin Q, Liu T, Qiao Y, Liu D, Yang L, Mao H, et al. Oxidative stress and inflammation in diabetic nephropathy: role of polyphenols. Front Immunol. 2023;14:1185317.
32. Calle P, Hotter G. Macrophage Phenotype and Fibrosis in Diabetic Nephropathy. Int J Mol Sci. 2020;21(8).
33. Liu Y, Xu K, Xiang Y, Ma B, Li H, Li Y, et al. Role of MCP-1 as an inflammatory biomarker in nephropathy. Front Immunol. 2023;14:1303076.
34. Rayego-Mateos S, Morgado-Pascual JL, Opazo-Ríos L, Guerrero-Hue M, García-Caballero C, Vázquez-Carballo C, et al. Pathogenic Pathways and Therapeutic Approaches Targeting Inflammation in Diabetic Nephropathy. Int J Mol Sci. 2020;21(11).
35. Guo W, Song Y, Sun Y, Du H, Cai Y, You Q, et al. Systemic immune-inflammation index is associated with diabetic kidney disease in Type 2 diabetes mellitus patients: Evidence from NHANES 2011-2018. Front Endocrinol (Lausanne). 2022;13:1071465.
36. Theocharis AD, Manou D, Karamanos NK. The extracellular matrix as a multitasking player in disease. FEBS J. 2019;286(15):2830-69.
37. Chaudhuri O, Cooper-White J, Janmey PA, Mooney DJ, Shenoy VB. Effects of extracellular matrix viscoelasticity on cellular behaviour. Nature. 2020;584(7822):535-46.
38. Li HD, You YK, Shao BY, Wu WF, Wang YF, Guo JB, et al. Roles and crosstalks of macrophages in diabetic nephropathy. Front Immunol. 2022;13:1015142.
39. Qin W, Cao L, Massey IY. Role of PI3K/Akt signaling pathway in cardiac fibrosis. Mol Cell Biochem. 2021;476(11):4045-59.
40. Wang J, Hu K, Cai X, Yang B, He Q, Wang J, et al. Targeting PI3K/AKT signaling for treatment of idiopathic pulmonary fibrosis. Acta pharmaceutica Sinica B. 2022;12(1):18-32.
41. Wang T, Wen X, Zhang Z, Xie M, Zhou J. Phillyrin ameliorates diabetic nephropathy through the PI3K/Akt/GSK-3β signalling pathway in streptozotocin-induced diabetic mice. Hum Exp Toxicol. 2021;40(12_suppl):S487-s96.
42. Wang J, Jiang C, Li N, Wang F, Xu Y, Shen Z, et al. The circEPSTI1/mir-942-5p/LTBP2 axis regulates the progression of OSCC in the background of OSF via EMT and the PI3K/Akt/mTOR pathway. Cell Death Dis. 2020;11(8):682.
43. Wu H, Xu F, Huang X, Li X, Yu P, Zhang L, et al. Lupenone improves type 2 diabetic nephropathy by regulating NF-κB pathway-mediated inflammation and TGF-β1/Smad/CTGF-associated fibrosis. Phytomedicine. 2023;118:154959.
44. Zou M, Zou J, Hu X, Zheng W, Zhang M, Cheng Z. Latent Transforming Growth Factor-β Binding Protein-2 Regulates Lung Fibroblast-to-Myofibroblast Differentiation in Pulmonary Fibrosis via NF-κB Signaling. Front Pharmacol. 2021;12:788714.
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| Diabetic nephropathy Inflammation Latent transforming growth factor-β binding protein-2 Mesangial cell Phosphatidylinositol 3-kinases/protein kinase B/nuclear factor kappa-B pathway | ||
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How to Cite
1.
Wang Y, Pi P, Hu M, Luo D. Inhibition of LTBP2 Suppresses High Glucose-Induced Proliferation, Fibrosis, and Inflammation in Glomerular Mesangial Cells by Disrupting the PI3K/Akt/NF-κB Pathway. Iran J Allergy Asthma Immunol. 2025;:1-14.
