The Role of Villin-1 in Model of Habu Nephritis Mice with Unilateral Nephrectomy

doi:10.3969/j.issn.1674-5817.2020.01.001

Abstract

Abstract: Objective To investigate the differences of renal function, renal pathological manifestations and proteins expression between the Habu nephritis mice model with or without unilateral nephrectomy (UNX), as well as the molecular mechanism leading to these differences. Methods Twenty- four male SPF C57BL/6 mice (18-20 g) at the age of 6 weeks were randomly divided into two groups. One group (n=12) received UNX and then received tail vein injection of Habu snake venom (HSV-UNX group) after 1 week, and the other group (n=12) received sham surgery and then received tail vein injection of Habu snake venom (HSV group). The renal function of mice in the two groups was detected by blood biochemistry, and the renal histopathological changes of two groups were measured through periodinate-schiff staining. Furthermore, the expression levels of protein in glomeruli of the two groups were analyzed by comparative proteomics, and the differentially expressed proteins between the two groups were screened out. The levels of proteins, such as villin-1(IL1), caspase 3, Bax, p21 and p27, were validated by Western blotting. Results The renal function injury of the HSV-UNX group was worse than that of the HSV group (P<0.01), and the renal mesangiolysis level was higher than that of the HSV group (P<0.0001), while the mesangial proliferation was milder than that of the HSV group (P<0.001). It is confirmed by proteomic analysis and western blotting that VIL1 expression level in the HSV-UNX group was lower than that of the HSV group. In mouse mesangial cells (MMCs), cell apoptosis (P<0.01) and inhibiting cell proliferation (P<0.05) were protomed by VIL1 knockdown through up-regulating the expressions of caspase 3, Bax, p21 and p27 proteins. Conclusions VIL1 is down-regulated in the HSV-UNX mouse model, which impairs the self-repair ability of glomerular and aggravates the renal function damage by inhibiting cell proliferation and promoting cell apoptosis.

Key words: Unilateral nephrectomy (UNX), Habu nephritis, Villin-1(VIL1), Mouse model

CLC Number:

Q95-33

JIANG Hongli, MA Hongye, XUE Jinhong, SUN Lingshuang, CHEN Lei. The Role of Villin-1 in Model of Habu Nephritis Mice with Unilateral Nephrectomy[J]. Laboratory Animal and Comparative Medicine, 2020, 40(1): 1-8.

References

[1] Gao J, Wu L, Wang Y, et al.Knockdown of Cxcl10 inhibits mesangial cell proliferation in murine Habu nephritis via ERK signaling[J]. Cell Physiol Biochem, 2017, 42(5):2118-2129.
[2] Jefferson JA, Johnson RJ .Experimental mesangial proliferative glomerulonephritis (the anti-Thy- 1.1 model)[J]. J Nephrol, 1999, 12(5):297-307.
[3] Emancipator SN .Prospects and perspectives on IgA nephropathy from animal models[J]. Contrib Nephrol, 2011, 169:126-152.
[4] Haas CS, Campean V, Kuhlmann A, et al.Analysis of glomerular VEGF mRNA and protein expression in murine mesangioproliferative glomerulonephritis[J]. Virchows Arch, 2007, 450(1):81-92.
[5] Nag S, Larsson M, Robinson RC, et al.Gelsolin: The tail of a molecular gymnast[J]. Cytoskeleton, 2013, 70(7):360-384.
[6] Lu Y, Cai G, Cui S, et al.FHL2-driven molecular network mediated Septin2 knockdown inducing apoptosis in mesangial cell[J]. Proteomics, 2014, 14(21-22):2485-2597.
[7] Abe-Yoshio Y, Abe K, Miyazaki M, et al.Involvement of bone marrow-derived endothelial progenitor cells in glomerular capillary repair in Habu snake venom-induced glomerulonephritis[J]. Virchows Arch, 2008, 453(1):97-106.
[8] He F, Zhou M, Yu T, et al.Sublytic C5b-9 triggers glomerular mesangial cell apoptosis in rat Thy-1 nephritis via Gadd45 activation mediated by Egr-1 and p300-dependent ATF3 acetylation[J].J Mol Cell Biol, 2016, 8(6):477-491.
[9] Blantz RC, Steiner RW.Benign hyperfiltration after living kidney donation[J]. J Clin Invest, 2015, 125(3):972-744.
[10] Wormser C, Aronson LR.Perioperative morbidity and long-term outcome of unilateral nephrectomy in feline kidney donors: 141 cases (1998-2013)[J]. J Am Vet Med Assoc, 2016, 248(3):275-281.
[11] Rayner HC, Ward L, Walls J, et al.Cholesterol feeding following unilateral nephrectomy in the rat leads to glomerular hypertrophy[J]. Nephron, 1991, 57:453-459.
[12] Uil M, Scantlebery AML, Butter LM, et al.Combining streptozotocin and unilateral nephrectomy is an effective method for inducing experimental diabetic nephropathy in the ‘resistant' C57Bl/6J mouse strain[J]. Sci Rep, 2018, 8(1):5542.
[13] Bretscher A, Weber K .Villin is a major protein of the microvillus cystoskeleton which binds both G and F actin in a calcium-dependent manner[J]. Cell, 1980, 20(3):839-847.
[14] Roy S, Esmaeilniakooshkghazi A, Patnaik S, et al.Villin-1 and gelsolin regulate changes in actin dynamics that affect cell survival signaling pathways and intestinal inflammation[J]. Gastroenterology, 2017, 154(5):1405-1420.
[15] Xieraili M, Yasen M, Mogushi K, et al.Villin 1 is a predictive factor for the recurrence of high serum alpha-fetoprotein-associated hepatocellular carcinoma after hepatectomy[J]. Cancer Sci , 2012, 103(8):1493-1501.
[16] Wang Y, Srinivasan K, Siddiqui MR, et al.A novel role for Villin in intestinal epithelial cell survival and homeostasis[J]. J Biol Chem, 2008, 283(14):9454-9464.
[17] Nakamura E, Satoh T, Iwakawa M, et al.Villin1, a diagnostic marker for endometrial adenocarcinoma with high grade nuclear atypia.[J]. Cancer Biol Ther, 2011, 12(3):181-190.
[18] Decuypere JP, Ceulemans LJ, Wylin T, et al.Plasmatic villin 1 is a novel in vivo marker of proximal tubular cell injury during renal ischemia-reperfusion[J]. Transplantation, 2017, 101(11):1-25.
[19] Jing X, Huang WH, Tang YJ, et al.Eucommia ulmoides Oliv. (Du-Zhong) lignans inhibit angiotensin II-stimulated proliferation by affecting p21, p27, and Bax expression in rat mesangial cells[J]. Evid Based Complement Alternat Med,2015, 2015(3):1-8.
[20] Ding Y, Wang Y, Chen J, et al.p21 overexpression sensitizes osteosarcoma U2OS cells to cisplatin via evoking caspase-3 and Bax/Bcl-2 cascade[J]. Tumour Biol, 2014, 35(4):3119-3123.
[21] Wang Y, George SP, Srinivasan K, et al. Reorganization of cytoskeleton as basis for apoptosis[J]. Gastroenterology, 2008, 134(4)(suppl 1):A-70.
[22] Meng J, Vardar D, Wang Y, et al.High-resolution crystal structures of villin headpiece and mutants with reduced F-actin binding activity[J]. Biochemistry, 2005, 44(36):11963-11973.
[23] Heijden MVD, Versteilen AMG, Sipkema P, et al.Rho-kinase-dependent F-actin rearrangement is involved in the inhibition of PI3-kinase/Akt during ischemia-reperfusion-induced endothelial cell apoptosis[J]. Apoptosis, 2008, 13(3):404-412.
[24] Lockett S, Verma C, Brafman A, et al.Quantitative analysis of F-actin redistribution in astrocytoma cells treated with candidate pharmaceuticals[J]. Cytometry A, 2014, 85(6):512-521.
[25] Ye JY, Liang EY, Cheng YS, et al.Serotonin enhances megakaryopoiesis and proplatelet formation via p-Erk1/2 and F-actin reorganization.[J]. Stem Cells, 2015, 32(11):2973-2982.