Early intervention with mesenchymal stem cells prevents nephropathy in diabetic rats by ameliorating the inflammatory microenvironment

Li,Yuanmin; Liu,Jingping; Liao,Guangneng; Zhang,Jie; Chen,Younan; Li,Lan; Li,Li; Liu,Fang; Chen,Bo; Guo,Gang; Wang,Chengshi; Yang,Lichuan; Cheng,Jingqiu; Lu,Yangrong

doi:10.3892/ijmm.2018.3501

Early intervention with mesenchymal stem cells prevents nephropathy in diabetic rats by ameliorating the inflammatory microenvironment

Authors:
- Yuanmin Li
- Jingping Liu
- Guangneng Liao
- Jie Zhang
- Younan Chen
- Lan Li
- Li Li
- Fang Liu
- Bo Chen
- Gang Guo
- Chengshi Wang
- Lichuan Yang
- Jingqiu Cheng
- Yangrong Lu
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Affiliations: Key Laboratory of Transplant Engineering and Immunology, National Health and Family Planning Commission (NHFPC), West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China, Department of Nephrology, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China
Published online on: February 16, 2018 https://doi.org/10.3892/ijmm.2018.3501
Pages: 2629-2639
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Diabetic nephropathy (DN) is a major complication of diabetes and represents the leading cause of end-stage renal disease. Mesenchymal stem cell (MSC) treatment has been demonstrated to be effective in DN models by reducing albuminuria and attenuating glomerular injury; however, limited in-depth understanding of the underlying mechanism and a lack of clinical trials hinders its clinical use. Additionally, most of these experimental studies were conducted on the advanced stage of nephropathy, which is difficult to reverse and consequently showed limited therapeutic efficacy. We sought to evaluate whether early intervention by MSCs has the potential to prevent DN onset and progression as well as protect kidney function when intravenously administered to rats with diabetes. Diabetes was induced in adult male SD rats by streptozotocin (STZ) injection (55 mg/kg, i.p.). The diabetic rats were injected with or without bone marrow-derived MSCs (5x106 per rat), via tail vein at 2, 4, 5 and 7 weeks after diabetes onset. Fasting blood glucose (FBG), blood urea nitrogen (BUN) and serum creatinine (Scr) levels in serum samples and glycosuria (GLU), microalbumin (MAU), and albumin to creatinine ratio (ACR) in urine samples were determined. Renal pathology and immunohistochemistry (IHC) for CD68, MCP-1, fibronectin (FN), transforming growth factor-β (TGF-β) and pro-inflammatory cytokines were also performed. Expression levels of the above factors as well as interleukin-10 (IL-10), and epidermal growth factor (EGF) were assessed by qPCR and multiplex bead-based suspension array system, respectively. Additionally, MSC tracing in vivo was performed. Ex vivo, peritoneal macrophages were co-cultured with MSCs, and expression of inflammatory cytokines was detected as well. MSC treatment profoundly suppressed renal macrophage infiltration and inflammatory cytokine secretion in diabetic rats, resulting in prominently improved kidney histology, systemic homeostasis, and animal survival, although no significant effect on hyperglycemia was observed. Engrafted MSCs were primarily localized in deteriorated areas of the kidney and immune organs 48 h after infusion. MSC treatment upregulated serum anti-inflammatory cytokines IL-10 and EGF. Ex vivo, MSCs inhibited lipopolysaccharide (LPS)-stimulated rat peritoneal macrophage activation via the downregulation of inflammatory-related cytokines such as IL-6, MCP-1, tumor necrosis factor-α (TNF-α) and IL-1β. Our results demonstrated that early intervention with MSCs prevented renal injury via immune regulation in diabetic rats, which restored the homeostasis of the immune microenvironment, contributing to the prevention of kidney dysfunction and glomerulosclerosis.

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1	Dronavalli S, Duka I and Bakris GL: The pathogenesis of diabetic nephropathy. Nat Clin Pract Endocrinol Metab. 4:444–452. 2008. View Article : Google Scholar : PubMed/NCBI
2	Maisonneuve P, Agodoa L, Gellert R, Stewart JH, Buccianti G, Lowenfels AB, Wolfe RA, Jones E, Disney APS, Briggs D, et al: Distribution of primary renal diseases leading to end-stage renal failure in the United States, Europe, and Australia/New Zealand: Results from an international comparative study. Am J Kidney Dis. 35:157–165. 2000. View Article : Google Scholar : PubMed/NCBI
3	Chavers BM, Bilous RW, Ellis EN, Steffes MW and Mauer SM: Glomerular lesions and urinary albumin excretion in type I diabetes without overt proteinuria. N Engl J Med. 320:966–970. 1989. View Article : Google Scholar : PubMed/NCBI
4	McCrary EB: The road to renal failure: An overview of diabetic nephropathy. Adv Nurse Pract. 16:61–63. 2008.
5	Schena FP and Gesualdo L: Pathogenetic mechanisms of diabetic nephropathy. J Am Soc Nephrol. 16(Suppl 1): S30–S33. 2005. View Article : Google Scholar : PubMed/NCBI
6	Navarro-González JF, Mora-Fernández C, Muros de Fuentes M and García-Pérez J: Inflammatory molecules and pathways in the pathogenesis of diabetic nephropathy. Nat Rev Nephrol. 7:327–340. 2011. View Article : Google Scholar : PubMed/NCBI
7	Navarro-González JF and Mora-Fernández C: The role of inflammatory cytokines in diabetic nephropathy. J Am Soc Nephrol. 19:433–442. 2008. View Article : Google Scholar : PubMed/NCBI
8	Cho DI, Kim MR, Jeong HY, Jeong HC, Jeong MH, Yoon SH, Kim YS and Ahn Y: Mesenchymal stem cells reciprocally regulate the M1/M2 balance in mouse bone marrow-derived macrophages. Exp Mol Med. 46:e702014. View Article : Google Scholar : PubMed/NCBI
9	Nguyen D, Ping F, Mu W, Hill P, Atkins RC and Chadban SJ: Macrophage accumulation in human progressive diabetic nephropathy. Nephrology (Carlton). 11:226–231. 2006. View Article : Google Scholar
10	Chow F, Ozols E, Nikolic-Paterson DJ, Atkins RC and Tesch GH: Macrophages in mouse type 2 diabetic nephropathy: Correlation with diabetic state and progressive renal injury. Kidney Int. 65:116–128. 2004. View Article : Google Scholar
11	Cao Q, Wang Y, Zheng D, Sun Y, Wang Y, Lee VWS, Zheng G, Tan TK, Ince J, Alexander SI, et al: IL-10/TGF-β-modified macrophages induce regulatory T cells and protect against adriamycin nephrosis. J Am Soc Nephrol. 21:933–942. 2010. View Article : Google Scholar : PubMed/NCBI
12	Prockop DJ and Oh JY: Mesenchymal stem/stromal cells (MSCs): Role as guardians of inflammation. Mol Ther. 20:14–20. 2012. View Article : Google Scholar :
13	Chow FY, Nikolic-Paterson DJ, Atkins RC and Tesch GH: Macrophages in streptozotocin-induced diabetic nephropathy: potential role in renal fibrosis. Nephrol Dial Transplant. 19:2987–2996. 2004. View Article : Google Scholar : PubMed/NCBI
14	Wang S, Li Y, Zhao J, Zhang J and Huang Y: Mesenchymal stem cells ameliorate podocyte injury and proteinuria in a type 1 diabetic nephropathy rat model. Biol Blood Marrow Transplant. 19:538–546. 2013. View Article : Google Scholar : PubMed/NCBI
15	Singer NG and Caplan AI: Mesenchymal stem cells: Mechanisms of inflammation. Annu Rev Pathol. 6:457–478. 2011. View Article : Google Scholar
16	Volarevic V, Arsenijevic N, Lukic ML and Stojkovic M: Concise review: Mesenchymal stem cell treatment of the complications of diabetes mellitus. Stem Cells. 29:5–10. 2011. View Article : Google Scholar : PubMed/NCBI
17	Lee RH, Seo MJ, Reger RL, Spees JL, Pulin AA, Olson SD and Prockop DJ: Multipotent stromal cells from human marrow home to and promote repair of pancreatic islets and renal glomeruli in diabetic NOD/scid mice. Proc Natl Acad Sci USA. 103:17438–17443. 2006. View Article : Google Scholar :
18	Tögel F, Hu Z, Weiss K, Isaac J, Lange C and Westenfelder C: Administered mesenchymal stem cells protect against ischemic acute renal failure through differentiation-independent mechanisms. Am J Physiol Renal Physiol. 289:F31–F42. 2005. View Article : Google Scholar : PubMed/NCBI
19	Tögel F, Weiss K, Yang Y, Hu Z, Zhang P and Westenfelder C: Vasculotropic, paracrine actions of infused mesenchymal stem cells are important to the recovery from acute kidney injury. Am J Physiol Renal Physiol. 292:F1626–F1635. 2007. View Article : Google Scholar : PubMed/NCBI
20	Ezquer FE, Ezquer ME, Parrau DB, Carpio D, Yañez AJ and Conget PA: Systemic administration of multipotent mesenchymal stromal cells reverts hyperglycemia and prevents nephropathy in type 1 diabetic mice. Biol Blood Marrow Transplant. 14:631–640. 2008. View Article : Google Scholar : PubMed/NCBI
21	Semedo P, Palasio CG, Oliveira CD, Feitoza CQ, Gonçalves GM, Cenedeze MA, Wang PMH, Teixeira VPA, Reis MA, Pacheco-Silva A, et al: Early modulation of inflammation by mesenchymal stem cell after acute kidney injury. Int Immunopharmacol. 9:677–682. 2009. View Article : Google Scholar : PubMed/NCBI
22	Semedo P, Correa-Costa M, Antonio Cenedeze M, Maria Avancini Costa Malheiros D, Antonia dos Reis M, Shimizu MH, Seguro AC, Pacheco-Silva A, Saraiva Camara NO and Olsen N: Mesenchymal stem cells attenuate renal fibrosis through immune modulation and remodeling properties in a rat remnant kidney model. Stem Cells. 27:3063–3073. 2009.PubMed/NCBI
23	Park JH, Hwang I, Hwang SH, Han H and Ha H: Human umbilical cord blood-derived mesenchymal stem cells prevent diabetic renal injury through paracrine action. Diabetes Res Clin Pract. 98:465–473. 2012. View Article : Google Scholar : PubMed/NCBI
24	Nakajima H, Uchida K, Guerrero AR, Watanabe S, Sugita D, Takeura N, Yoshida A, Long G, Wright KT, Johnson WEB, et al: Transplantation of mesenchymal stem cells promotes an alternative pathway of macrophage activation and functional recovery after spinal cord injury. J Neurotrauma. 29:1614–1625. 2012. View Article : Google Scholar : PubMed/NCBI
25	Jiang TS, Cai L, Ji WY, Hui YN, Wang YS, Hu D and Zhu J: Reconstruction of the corneal epithelium with induced marrow mesenchymal stem cells in rats. Mol Vis. 16:1304–1316. 2010.PubMed/NCBI
26	Mafi P: Adult mesenchymal stem cells and cell surface characterization - a systematic review of the literature. Open Orthop J. 5(Suppl 2): 253–260. 2011. View Article : Google Scholar
27	Zhang X, Goncalves R and Mosser DM: The isolation and characterization of murine macrophages. Curr Protoc Immunol. Chapter: Unit-14.1. 2008. View Article : Google Scholar
28	Wang Y, Wang Y, Feng X, Bao S, Yi S, Kairaitis L, Tay YC, Rangan GK and Harris DC: Depletion of CD4(+) T cells aggravates glomerular and interstitial injury in murine adriamycin nephropathy. Kidney Int. 59:975–984. 2001. View Article : Google Scholar : PubMed/NCBI
29	Kim J and Hematti P: Mesenchymal stem cell-educated macrophages: A novel type of alternatively activated macrophages. Exp Hematol. 37:1445–1453. 2009. View Article : Google Scholar : PubMed/NCBI
30	Tesch GH and Allen TJ: Rodent models of streptozotocin-induced diabetic nephropathy. Nephrology (Carlton). 12:261–266. 2007. View Article : Google Scholar
31	Liu CX, Hu Q, Wang Y, Zhang W, Ma ZY, Feng JB, Wang R, Wang XP, Dong B, Gao F, et al: Angiotensin-converting enzyme (ACE) 2 overexpression ameliorates glomerular injury in a rat model of diabetic nephropathy: A comparison with ACE inhibition. Mol Med. 17:59–69. 2011.
32	Lv SS, Liu G, Wang JP, Wang WW, Cheng J, Sun AL, Liu HY, Nie HB, Su MR and Guan GJ: Mesenchymal stem cells transplantation ameliorates glomerular injury in streptozotocin-induced diabetic nephropathy in rats via inhibiting macrophage infiltration. Int Immunopharmacol. 17:275–282. 2013. View Article : Google Scholar : PubMed/NCBI
33	Lv S, Cheng J, Sun A, Li J, Wang W, Guan G, Liu G and Su M: Mesenchymal stem cells transplantation ameliorates glomerular injury in streptozotocin-induced diabetic nephropathy in rats via inhibiting oxidative stress. Diabetes Res Clin Pract. 104:143–154. 2014. View Article : Google Scholar : PubMed/NCBI
34	Tesch GH: MCP-1/CCL2: A new diagnostic marker and therapeutic target for progressive renal injury in diabetic nephropathy. Am J Physiol Renal Physiol. 294:F697–F701. 2008. View Article : Google Scholar : PubMed/NCBI
35	Brady HR: Leukocyte adhesion molecules and kidney diseases. Kidney Int. 45:1285–1300. 1994. View Article : Google Scholar : PubMed/NCBI
36	Duffield JS, Erwig L-P, Wei X, Liew FY, Rees AJ and Savill JS: Activated macrophages direct apoptosis and suppress mitosis of mesangial cells. J Immunol. 164:2110–2119. 2000. View Article : Google Scholar : PubMed/NCBI
37	Chow FY, Nikolic-Paterson DJ, Ozols E, Atkins RC and Tesch GH: Intercellular adhesion molecule-1 deficiency is protective against nephropathy in type 2 diabetic db/db mice. J Am Soc Nephrol. 16:1711–1722. 2005. View Article : Google Scholar : PubMed/NCBI
38	Banba N, Nakamura T, Matsumura M, Kuroda H, Hattori Y and Kasai K: Possible relationship of monocyte chemoattractant protein-1 with diabetic nephropathy. Kidney Int. 58:684–690. 2000. View Article : Google Scholar : PubMed/NCBI
39	Santos JC, de Brito CA, Futata EA, Azor MH, Orii NM, Maruta CW, Rivitti EA, Duarte AJS and Sato MN: Upregulation of chemokine C-C ligand 2 (CCL2) and C-X-C chemokine 8 (CXCL8) expression by monocytes in chronic idiopathic urticaria. Clin Exp Immunol. 167:129–136. 2012. View Article : Google Scholar :
40	Li W, Zhang Q, Wang M, Wu H, Mao F, Zhang B, Ji R, Gao S, Sun Z, Zhu W, et al: Macrophages are involved in the protective role of human umbilical cord-derived stromal cells in renal ischemia-reperfusion injury. Stem Cell Res (Amst). 10:405–416. 2013. View Article : Google Scholar
41	Villanueva S, Ewertz E, Carrión F, Tapia A, Vergara C, Céspedes C, Sáez PJ, Luz P, Irarrázabal C, Carreño JE, et al: Mesenchymal stem cell injection ameliorates chronic renal failure in a rat model. Clin Sci (Lond). 121:489–499. 2011. View Article : Google Scholar
42	Ezquer F, Giraud-Billoud M, Carpio D, Cabezas F, Conget P and Ezquer M: Proregenerative microenvironment triggered by donor mesenchymal stem cells preserves renal function and structure in mice with severe diabetes mellitus. Biomed Res Int. 2015:1647032015. View Article : Google Scholar : PubMed/NCBI
43	Bi B, Schmitt R, Israilova M, Nishio H and Cantley LG: Stromal cells protect against acute tubular injury via an endocrine effect. J Am Soc Nephrol. 18:2486–2496. 2007. View Article : Google Scholar : PubMed/NCBI
44	Zhang A, Wang Y, Ye Z, Xie H, Zhou L and Zheng S: Mechanism of TNF-α-induced migration and hepatocyte growth factor production in human mesenchymal stem cells. J Cell Biochem. 111:469–475. 2010. View Article : Google Scholar : PubMed/NCBI
45	Kim YH, Ryu JM, Lee YJ and Han HJ: Fibronectin synthesis by high glucose level mediated proliferation of mouse embryonic stem cells: Involvement of ANG II and TGF-β1. J Cell Physiol. 223:397–407. 2010.PubMed/NCBI
46	Wise AF and Ricardo SD: Mesenchymal stem cells in kidney inflammation and repair. Nephrology (Carlton). 17:1–10. 2012. View Article : Google Scholar
47	Rasmusson I: Immune modulation by mesenchymal stem cells. Exp Cell Res. 312:2169–2179. 2006. View Article : Google Scholar : PubMed/NCBI
48	Meisel R, Zibert A, Laryea M, Göbel U, Däubener W and Dilloo D: Human bone marrow stromal cells inhibit allogeneic T-cell responses by indoleamine 2,3-dioxygenase-mediated tryptophan degradation. Blood. 103:4619–4621. 2004. View Article : Google Scholar : PubMed/NCBI
49	Tipnis S, Viswanathan C and Majumdar AS: Immunosuppressive properties of human umbilical cord-derived mesenchymal stem cells: Role of B7-H1 and IDO. Immunol Cell Biol. 88:795–806. 2010. View Article : Google Scholar : PubMed/NCBI