Urinary miR-29 correlates with albuminuria and carotid intima-media thickness in type 2 diabetes patients

Hui Peng, Meirong Zhong, Wenbo Zhao, Cheng Wang, Jun Zhang, Xun Liu, Yuanqing Li, Sujay Dutta Paudel, Qianqian Wang, Tanqi Lou, Hui Peng, Meirong Zhong, Wenbo Zhao, Cheng Wang, Jun Zhang, Xun Liu, Yuanqing Li, Sujay Dutta Paudel, Qianqian Wang, Tanqi Lou

Abstract

Background: Cell-free microRNAs stably and abundantly exist in body fluids and emerging evidence suggests cell-free microRNAs as novel and non-invasive disease biomarker. Deregulation of miR-29 is involved in the pathogenesis of diabetic nephropathy and insulin resistance thus may be implicated in diabetic vascular complication. Therefore, we investigated the possibility of urinary miR-29 as biomarker for diabetic nephropathy and atherosclerosis in patients with type 2 diabetes.

Methods: 83 patients with type 2 diabetes were enrolled in this study, miR-29a, miR-29b and miR-29c levels in urine supernatant was determined by TaqMan qRT-PCR, and a synthetic cel-miR-39 was added to the urine as a spike-in control before miRNAs extraction. Urinary albumin excretion rate and urine albumin/creatinine ratio, funduscopy and carotid ultrasound were used for evaluation of diabetic vascular complication. The laboratory parameters indicating blood glucose level, renal function and serum lipids were also collected.

Results: Patients with albuminuria (n = 42, age 60.62 ± 12.00 yrs) showed significantly higher comorbidity of diabetic retinopathy (p = 0.015) and higher levels of urinary miR-29a (p = 0.035) compared with those with normoalbuminuria (n = 41, age 58.54 ± 14.40 yrs). There was no significant difference in urinary miR-29b (p = 0.148) or miR-29c level (p = 0.321) between groups. Urinary albumin excretion rate significantly correlated with urinary miR-29a level (r = 0.286, p = 0.016), while urinary miR-29b significantly correlated with carotid intima-media thickness (cIMT) (r = 0.286, p = 0.046).

Conclusion: Urinary miR-29a correlated with albuminuria while urinary miR-29b correlated with carotid intima-media thickness (cIMT) in patients with type 2 diabetes. Therefore, they may have the potential to serve as alternative biomarker for diabetic nephropathy and atherosclerosis in type 2 diabetes.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1. The relative abundance of urinary…
Figure 1. The relative abundance of urinary miR-29 members in patients with Type 2 diabetes mellitus (n = 83).
Urinary miR-29a and miR-29c are significantly higher than miR-29b in type 2 diabetes patients (both with a p value

Figure 2. Comparison of urinary miR-29 members…

Figure 2. Comparison of urinary miR-29 members between diabetes patients with albuminuria and normoalbuminuria.

Urinary…

Figure 2. Comparison of urinary miR-29 members between diabetes patients with albuminuria and normoalbuminuria.
Urinary miR-29a was higher in diabetes with albuminuria group than in diabetes with normoalbuminuria group (p = 0.035). There was no significant difference in urinary miR-29b (p = 0.148) or miR-29c (p = 0.321) levels between two groups. The values are represented as ratio to the median of diabetes with normoalbuminuria group. Data are compared by Mann-Whitney U test.

Figure 3. Correlation between urinary miR-29 members…

Figure 3. Correlation between urinary miR-29 members and urinary albumin excretion rate.

a: Urinary miR-29a…

Figure 3. Correlation between urinary miR-29 members and urinary albumin excretion rate.
a: Urinary miR-29a significantly correlated with urinary albumin excretion rate (r = 0.286, p = 0.016). b: Correlation between urinary albumin excretion rate and miR-29b was borderline significant (r = 0.212, p = 0.078). c: There was no significant correlation between miR-29c and urinary albumin excretion rate (r = 0.151, p = 0.211). Data were compared by Spearman’s rank order correlations.

Figure 4. Correlation between urinary miR-29 members…

Figure 4. Correlation between urinary miR-29 members and carotid intima-media thickness (cIMT).

a: There was…

Figure 4. Correlation between urinary miR-29 members and carotid intima-media thickness (cIMT).
a: There was no significant correlation between urinary miR-29a and carotid intima-media thickness (cIMT) (r = 0.173, p = 0.234). b: Urinary miR-29b significantly correlated with cIMT (r = 0.286, p = 0.046). c: There was no significant correlation between urinary miR-29c and cIMT (r = 0.048, p = 0.741) levels. Data were compared by Spearman’s rank order correlations.
Figure 2. Comparison of urinary miR-29 members…
Figure 2. Comparison of urinary miR-29 members between diabetes patients with albuminuria and normoalbuminuria.
Urinary miR-29a was higher in diabetes with albuminuria group than in diabetes with normoalbuminuria group (p = 0.035). There was no significant difference in urinary miR-29b (p = 0.148) or miR-29c (p = 0.321) levels between two groups. The values are represented as ratio to the median of diabetes with normoalbuminuria group. Data are compared by Mann-Whitney U test.
Figure 3. Correlation between urinary miR-29 members…
Figure 3. Correlation between urinary miR-29 members and urinary albumin excretion rate.
a: Urinary miR-29a significantly correlated with urinary albumin excretion rate (r = 0.286, p = 0.016). b: Correlation between urinary albumin excretion rate and miR-29b was borderline significant (r = 0.212, p = 0.078). c: There was no significant correlation between miR-29c and urinary albumin excretion rate (r = 0.151, p = 0.211). Data were compared by Spearman’s rank order correlations.
Figure 4. Correlation between urinary miR-29 members…
Figure 4. Correlation between urinary miR-29 members and carotid intima-media thickness (cIMT).
a: There was no significant correlation between urinary miR-29a and carotid intima-media thickness (cIMT) (r = 0.173, p = 0.234). b: Urinary miR-29b significantly correlated with cIMT (r = 0.286, p = 0.046). c: There was no significant correlation between urinary miR-29c and cIMT (r = 0.048, p = 0.741) levels. Data were compared by Spearman’s rank order correlations.

References

    1. Cooper ME, Gilbert RE, Jerums G (1997) Diabetic vascular complications. Clin Exp Pharmacol Physiol 24: 770–775.
    1. Rosenson RS, Fioretto P, Dodson PM (2011) Does microvascular disease predict macrovascular events in type 2 diabetes? Atherosclerosis 218: 13–18.
    1. Tan SM, Sharma A, Yuen DY, Stefanovic N, Krippner G, et al. (2013) The Modified Selenenyl Amide, M-hydroxy Ebselen, Attenuates Diabetic Nephropathy and Diabetes-Associated Atherosclerosis in ApoE/GPx1 Double Knockout Mice. PLoS One 8: e69193.
    1. McKittrick IB, Bogaert Y, Nadeau K, Snell-Bergeon J, Hull A, et al. (2011) Urinary matrix metalloproteinase activities: biomarkers for plaque angiogenesis and nephropathy in diabetes. Am J Physiol Renal Physiol 301: F1326–F1333.
    1. Esteller M (2011) Non-coding RNAs in human disease. Nat Rev Genet 12: 861–874.
    1. Natarajan R, Putta S, Kato M (2012) MicroRNAs and diabetic complications. J Cardiovasc Transl Res 5: 413–422.
    1. Chen X, Ba Y, Ma L, Cai X, Yin Y, et al. (2008) Characterization of microRNAs in serum: a novel class of biomarkers for diagnosis of cancer and other diseases. Cell Res 18: 997–1006.
    1. Wang K, Zhang S, Weber J, Baxter D, Galas DJ (2010) Export of microRNAs and microRNA-protective protein by mammalian cells. Nucleic Acids Res 38: 7248–7259.
    1. Arroyo JD, Chevillet JR, Kroh EM, Ruf IK, Pritchard CC, et al. (2011) Argonaute2 complexes carry a population of circulating microRNAs independent of vesicles in human plasma. Proc Natl Acad Sci U S A 108: 5003–5008.
    1. Vickers KC, Palmisano BT, Shoucri BM, Shamburek RD, Remaley AT (2011) MicroRNAs are transported in plasma and delivered to recipient cells by high-density lipoproteins. Nat Cell Biol 13: 423–433.
    1. He Y, Huang C, Lin X, Li J (2013) MicroRNA-29 family, a crucial therapeutic target for fibrosis diseases. Biochimie 95: 1355–1359.
    1. Long J, Wang Y, Wang W, Chang BH, Danesh FR (2011) MicroRNA-29c is a signature microRNA under high glucose conditions that targets Sprouty homolog 1, and its in vivo knockdown prevents progression of diabetic nephropathy. J Biol Chem 286: 11837–11848.
    1. Wang B, Komers R, Carew R, Winbanks CE, Xu B, et al. (2012) Suppression of microRNA-29 expression by TGF-beta1 promotes collagen expression and renal fibrosis. J Am Soc Nephrol 23: 252–265.
    1. Rask-Madsen C, King GL (2013) Vascular complications of diabetes: mechanisms of injury and protective factors. Cell Metab 17: 20–33.
    1. He A, Zhu L, Gupta N, Chang Y, Fang F (2007) Overexpression of micro ribonucleic acid 29, highly up-regulated in diabetic rats, leads to insulin resistance in 3T3-L1 adipocytes. Mol Endocrinol 21: 2785–2794.
    1. Welsh GI, Hale LJ, Eremina V, Jeansson M, Maezawa Y, et al. (2010) Insulin signaling to the glomerular podocyte is critical for normal kidney function. Cell Metab 12: 329–340.
    1. Rask-Madsen C, Li Q, Freund B, Feather D, Abramov R, et al. (2010) Loss of insulin signaling in vascular endothelial cells accelerates atherosclerosis in apolipoprotein E null mice. Cell Metab 11: 379–389.
    1. Lupattelli G, Vuono SD, Boni M, Helou R, Mannarino MR, et al. (2013) Insulin Resistance and not BMI is the Major Determinant of Early Vascular Impairment in Patients with Morbid Obesity. J Atheroscler Thromb 21: 2785–2794.
    1. Kanter JE, Bornfeldt KE (2013) Evidence stacks up that endothelial insulin resistance is a culprit in atherosclerosis. Circ Res 113: 352–354.
    1. Cheng Y, Wang X, Yang J, Duan X, Yao Y, et al. (2012) A translational study of urine miRNAs in acute myocardial infarction. J Mol Cell Cardiol 53: 668–676.
    1. Levey AS, Stevens LA, Schmid CH, Zhang YL, Castro AR, et al. (2009) A new equation to estimate glomerular filtration rate. Ann Intern Med 150: 604–612.
    1. Ferrannini E, Cushman WC (2012) Diabetes and hypertension: the bad companions. Lancet 380: 601–610.
    1. Lou QL, Ouyang XJ, Gu LB, Mo YZ, Ma R, et al. (2012) Chronic kidney disease and associated cardiovascular risk factors in chinese with type 2 diabetes. Diabetes Metab J 36: 433–442.
    1. He F, Xia X, Wu XF, Yu XQ, Huang FX (2013) Diabetic retinopathy in predicting diabetic nephropathy in patients with type 2 diabetes and renal disease: a meta-analysis. Diabetologia 56: 457–466.
    1. Chen YH, Chen HS, Tarng DC (2012) More impact of microalbuminuria on retinopathy than moderately reduced GFR among type 2 diabetic patients. Diabetes Care 35: 803–808.
    1. Kramer CK, Retnakaran R (2013) Concordance of retinopathy and nephropathy over time in Type 1 diabetes: an analysis of data from the Diabetes Control and Complications Trial. Diabet Med 35: 803–808.
    1. Kriegel AJ, Liu Y, Fang Y, Ding X, Liang M (2012) The miR-29 family: genomics, cell biology, and relevance to renal and cardiovascular injury. Physiol Genomics 44: 237–244.
    1. Fang Y, Yu X, Liu Y, Kriegel AJ, Heng Y, et al. (2013) miR-29c is downregulated in renal interstitial fibrosis in humans and rats and restored by HIF-alpha activation. Am J Physiol Renal Physiol 304: F1274–F1282.
    1. Wang G, Kwan BC, Lai FM, Chow KM, Li PK, et al. (2012) Urinary miR-21, miR-29, and miR-93: novel biomarkers of fibrosis. Am J Nephrol 36: 412–418.
    1. Lv LL, Cao YH, Ni HF, Xu M, Liu D, et al. (2013) microRNA-29c in urinary exosome/microvesicle as biomarker of renal fibrosis. Am J Physiol Renal Physiol 36: 412–418.
    1. Hanke M, Hoefig K, Merz H, Feller AC, Kausch I, et al. (2010) A robust methodology to study urine microRNA as tumor marker: microRNA-126 and microRNA-182 are related to urinary bladder cancer. Urol Oncol 28: 655–661.
    1. Zen K, Zhang CY (2012) Circulating microRNAs: a novel class of biomarkers to diagnose and monitor human cancers. Med Res Rev 32: 326–348.
    1. Wang G, Chan ES, Kwan BC, Li PK, Yip SK, et al. (2012) Expression of microRNAs in the urine of patients with bladder cancer. Clin Genitourin Cancer 10: 106–113.
    1. van Dam EM, Govers R, James DE (2005) Akt activation is required at a late stage of insulin-induced GLUT4 translocation to the plasma membrane. Mol Endocrinol 19: 1067–1077.
    1. Brozinick JJ, Roberts BR, Dohm GL (2003) Defective signaling through Akt-2 and -3 but not Akt-1 in insulin-resistant human skeletal muscle: potential role in insulin resistance. Diabetes 52: 935–941.
    1. Wei W, He HB, Zhang WY, Zhang HX, Bai JB, et al. (2013) miR-29 targets Akt3 to reduce proliferation and facilitate differentiation of myoblasts in skeletal muscle development. Cell Death Dis 4: e668.
    1. Muniyappa R, Montagnani M, Koh KK, Quon MJ (2007) Cardiovascular actions of insulin. Endocr Rev 28: 463–491.
    1. Rask-Madsen C, Li Q, Freund B, Feather D, Abramov R, et al. (2010) Loss of insulin signaling in vascular endothelial cells accelerates atherosclerosis in apolipoprotein E null mice. Cell Metab 11: 379–389.

Source: PubMed

Sponsorit ja yhteistyökumppanit

Sairaudet

Huumeiden interventiot

3
Tilaa