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Urinary Cyclophilin A as a New Marker for Diabetic Nephropathy: A Cross-Sectional Analysis of Diabetes Mellitus.

Tsai SF, Su CW, Wu MJ, Chen CH, Fu CP, Liu CS, Hsieh M - Medicine (Baltimore) (2015)

Bottom Line: A significant increase in urinary CypA was noted in stage 2 DN and persisted in later stages.The area under curve was up to 0.85, indicating a good discriminatory power.The underlying molecular mechanisms still need further clarification in cellular and animal studies.

View Article: PubMed Central - PubMed

Affiliation: From the Division of Nephrology, Department of Internal Medicine, Taichung Veterans General Hospital (S-FT, M-JW, C-HC); School of Medicine, China Medical University (S-FT, C-HC); Department of Life Science, Tunghai University (S-FT, C-WS, C-HC, MH); School of Medicine, Chung Shan Medical University (M-JW, C-HC); Division of Endocrinology and Metabolism, Department of Medicine, Taichung Veterans General Hospital, Taichung (C-PF); Vascular and Genomic Research Center, Changhua Christian Hospital, Changhua (C-SL); and Life Science Research Center, Tunghai University, Taichung, Taiwan R.O.C. (MH).

ABSTRACT
Type 2 diabetes mellitus (DM) is the most common single cause of end-stage renal disease. Albuminuria is the most commonly used marker to predict onset of diabetic nephropathy (DN) without enough sensitivity and specificity to detect early DN. This is the first study to identify urinary cyclophilin A (CypA) as a new biomarker for early DN.We recruited DM outpatients and healthy control subjects from January 2014 to December 2014. In this cross-sectional study, patients' urine samples were collected to determine the expression of urinary CypA. We also treated mesangial (MES-13) and tubular (HK-2) cells with glucose or free radicals to observe the expression of secreted CypA in Western blot analysis.A total of 100 DN patients and 20 healthy control subjects were enrolled. All variables were matched. In univariate analysis, the concentration of urinary CypA correlated well with the progression of renal function. A significant increase in urinary CypA was noted in stage 2 DN and persisted in later stages. We could diagnose stage 2 DN using urinary CypA with a sensitivity of 90.0% and specificity of 72.7%. The area under curve was up to 0.85, indicating a good discriminatory power. In cellular models, MES-13 and HK-2 cells can both release CypA.Urinary CypA is a good biomarker for early DN detection in humans and it can be released from either mesangial or tubular cells. The underlying molecular mechanisms still need further clarification in cellular and animal studies.

No MeSH data available.


Related in: MedlinePlus

Western blotting of sCypA expression in HK-2 cells treated with different concentrations of glucose and H2O2. (A) HK-2 cells were treated with different concentrations (10, 25, and 50 mM) of glucose. All concentrations of glucose increased the expression of sCypA. (B) The expression of sCypA was increased in cells with 20 or 40 μM H2O2 treatments. They could both be counteracted by 300 U/mL catalase. Endo-CypA = endogenous cyclophilin A, sCypA = secreted cyclophilin A.
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Figure 5: Western blotting of sCypA expression in HK-2 cells treated with different concentrations of glucose and H2O2. (A) HK-2 cells were treated with different concentrations (10, 25, and 50 mM) of glucose. All concentrations of glucose increased the expression of sCypA. (B) The expression of sCypA was increased in cells with 20 or 40 μM H2O2 treatments. They could both be counteracted by 300 U/mL catalase. Endo-CypA = endogenous cyclophilin A, sCypA = secreted cyclophilin A.

Mentions: Mesangial cell injury is the classical expression of DN, but recent studies suggested that DN is also a tubular disease. Early changes in tubular epithelial cells may be an essential factor in the development of progressive kidney diseases.34 HK-2 cells, human PTEC, have been used as a cell model to study tubular diseases. Therefore, Western blotting was used to disclose the expression of sCypA whereby various concentrations of glucose and H2O2 were applied to HK-2 cells. As expected, different concentrations of glucose (10, 25, and 50 mM) could effectively increase the expression of sCypA (Fig. 5A), indicating that hyperglycemia can also induce sCypA release from tubular cells. In addition, either 20 or 40 μM H2O2 treatment significantly increased the expression of sCypA, which could be reversed by 300 U/mL of catalase (Fig. 5B).


Urinary Cyclophilin A as a New Marker for Diabetic Nephropathy: A Cross-Sectional Analysis of Diabetes Mellitus.

Tsai SF, Su CW, Wu MJ, Chen CH, Fu CP, Liu CS, Hsieh M - Medicine (Baltimore) (2015)

Western blotting of sCypA expression in HK-2 cells treated with different concentrations of glucose and H2O2. (A) HK-2 cells were treated with different concentrations (10, 25, and 50 mM) of glucose. All concentrations of glucose increased the expression of sCypA. (B) The expression of sCypA was increased in cells with 20 or 40 μM H2O2 treatments. They could both be counteracted by 300 U/mL catalase. Endo-CypA = endogenous cyclophilin A, sCypA = secreted cyclophilin A.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4620809&req=5

Figure 5: Western blotting of sCypA expression in HK-2 cells treated with different concentrations of glucose and H2O2. (A) HK-2 cells were treated with different concentrations (10, 25, and 50 mM) of glucose. All concentrations of glucose increased the expression of sCypA. (B) The expression of sCypA was increased in cells with 20 or 40 μM H2O2 treatments. They could both be counteracted by 300 U/mL catalase. Endo-CypA = endogenous cyclophilin A, sCypA = secreted cyclophilin A.
Mentions: Mesangial cell injury is the classical expression of DN, but recent studies suggested that DN is also a tubular disease. Early changes in tubular epithelial cells may be an essential factor in the development of progressive kidney diseases.34 HK-2 cells, human PTEC, have been used as a cell model to study tubular diseases. Therefore, Western blotting was used to disclose the expression of sCypA whereby various concentrations of glucose and H2O2 were applied to HK-2 cells. As expected, different concentrations of glucose (10, 25, and 50 mM) could effectively increase the expression of sCypA (Fig. 5A), indicating that hyperglycemia can also induce sCypA release from tubular cells. In addition, either 20 or 40 μM H2O2 treatment significantly increased the expression of sCypA, which could be reversed by 300 U/mL of catalase (Fig. 5B).

Bottom Line: A significant increase in urinary CypA was noted in stage 2 DN and persisted in later stages.The area under curve was up to 0.85, indicating a good discriminatory power.The underlying molecular mechanisms still need further clarification in cellular and animal studies.

View Article: PubMed Central - PubMed

Affiliation: From the Division of Nephrology, Department of Internal Medicine, Taichung Veterans General Hospital (S-FT, M-JW, C-HC); School of Medicine, China Medical University (S-FT, C-HC); Department of Life Science, Tunghai University (S-FT, C-WS, C-HC, MH); School of Medicine, Chung Shan Medical University (M-JW, C-HC); Division of Endocrinology and Metabolism, Department of Medicine, Taichung Veterans General Hospital, Taichung (C-PF); Vascular and Genomic Research Center, Changhua Christian Hospital, Changhua (C-SL); and Life Science Research Center, Tunghai University, Taichung, Taiwan R.O.C. (MH).

ABSTRACT
Type 2 diabetes mellitus (DM) is the most common single cause of end-stage renal disease. Albuminuria is the most commonly used marker to predict onset of diabetic nephropathy (DN) without enough sensitivity and specificity to detect early DN. This is the first study to identify urinary cyclophilin A (CypA) as a new biomarker for early DN.We recruited DM outpatients and healthy control subjects from January 2014 to December 2014. In this cross-sectional study, patients' urine samples were collected to determine the expression of urinary CypA. We also treated mesangial (MES-13) and tubular (HK-2) cells with glucose or free radicals to observe the expression of secreted CypA in Western blot analysis.A total of 100 DN patients and 20 healthy control subjects were enrolled. All variables were matched. In univariate analysis, the concentration of urinary CypA correlated well with the progression of renal function. A significant increase in urinary CypA was noted in stage 2 DN and persisted in later stages. We could diagnose stage 2 DN using urinary CypA with a sensitivity of 90.0% and specificity of 72.7%. The area under curve was up to 0.85, indicating a good discriminatory power. In cellular models, MES-13 and HK-2 cells can both release CypA.Urinary CypA is a good biomarker for early DN detection in humans and it can be released from either mesangial or tubular cells. The underlying molecular mechanisms still need further clarification in cellular and animal studies.

No MeSH data available.


Related in: MedlinePlus