Abstract: TH-PO382

Increased Expression of Soluble Fas in Patients with CKD

Session Information

Category: Cell Biology

  • 201 Cell Signaling, Oxidative Stress

Authors

  • Goes, Miguel A, Hospital Israelita Albert Einstein, Sao Paulo, Brazil
  • Canziani, Maria Eugenia F., Federal University of Sao Paulo, Sao Paulo, Brazil
  • Manfredi, Silvia Regina, Universidade Federal de São Paulo , São Paulo, Brazil
  • Dalboni, Maria, Universidade Federal de São Paulo , São Paulo, Brazil
Background

Fas (CD95) is a cellular receptor for apoptosis in leukocytes and other cells. A soluble form of Fas (sFas) is an anti-apoptotic molecule devoid of the transmembrane domain from alternative splicing of CD95. Serum sFas levels are higher in CKD patients and associated with inflammation, anemia, and cardiovascular disease. Objective: To investigate whether the expression of CD95mRNA and sFasmRNA is increase in leukocytes of CKD patients and their respective correlation with serum soluble Fas levels.

Methods

We performed the dosage of Hb concentration, serum creatinine and urea by conventional methods and serum sFas levels measured using an enzyme-linked immunosorbent assay from 51 CKD patients (eGFR 15 to 59 ml/min; CKD group) and 18 healthy volunteers (control group). We extracted leukocytes to measure the mRNA expression of CD95 and sFas. Total RNA was isolated from 5 x 106 leukocytes from each subject using TRIzol reagent, and cDNA was synthesized from 1 μg RNA using reverse transcriptase synthesis system. Relative levels of mRNA transcripts of sFas were quantified by real-time PCR. We used the Epi-CKD formula. We perform correlations and comparisons between groups.

Results

When analyzed both groups together we observed negative correlation between eGFR serum sFas levels (r=-0.30, p=0.01), between eGFR and sFasmRNA expression (r= - 0.28, p= 0.02). Serum sFas levels correlated positively with sFasmRNA copies (r = 0.32, p = 0.007). The CD95mRNA did not correlate with eGFR (r= - 0.04, p= 0.9).
The main etiologies of CKD were diabetes and hypertension. We observed lower concentration of Hb in the CKD group (10.8 + 2.1, 14.2 +1.7, p <0.001). There was a higher serum sFas levels in CKD group (3161+ 1000, 1686+ 996, p <0.001) and higher copies of sFasmRNA in the CKD group (32.3 + 2.3 x 106, 23.3 + 5.9 x 106, p <0.001). There was a negative correlation between CD95mRNA and sFasmRNA copies in CKD patients (r= - 0.49, p <0.001).

Conclusion

Serum sFas levels and sFasmRNA expression are elevated in patients with CKD. We observed correlation between sFasmRNA expression and serum levels of sFas