Expression of lncRNA GAS5-AS1 and IL-27 in peripheral blood of patients with primary membranous nephropathy and their relationship with Treg/Th17 balance

Objective  To detect the expression of long non-coding RNA growth arrest-specific transcript 5 antisense RNA1(lncRNA GAS5-AS1) and interleukin(IL)-27 in peripheral blood of patients with primary membranous nephropathy(PMN), and to analyze their relationship with regulatory T cell/T help 17 cells(Treg/Th17) balance in patients.

Methods  Ninety-four PMN patients admitted to our hospital from September 2019 to September 2020 were regarded as the PMN group. Another 96 cases with normal physical examination during the same course were regarded as the control group. Peripheral blood samples were collected from all study subjects, real-time quantitative PCR(qRT-PCR) method was applied to determine the serum lncRNA GAS5-AS1 level, enzyme-linked immunosorbent assay(ELISA) was used to measure the levels of IL-27, IL-17, tumor necrosis factor-α(TNF-α), IL-10, transforming growth factor betal(TGF-β1), flow cytometry was used to determine the proportions of Treg cells and Th17 cells in peripheral blood, and to calculate the Treg/Th17 ratio. Pearson method was applied to analyze the correlation of lncRNA GAS5-AS1 and IL-27 levels with IL-17, TNF-α, IL-10, TGF-β1, and Treg/Th17 in PMN patients.

Results Compared with the control group, blood urea nitrogen(BUN)(6.52±1.05) mmol/L vs (4.03±0.66) mmol/L, uric acid(UA)(338.92±56.45) μmol/L vs (266.13±44.35) μmol/L, cystatin C(Cys-C)(1.38±0.23) mg/L vs (0.72±0.12) mg/L, phospholipase A2 receptor antibody(PLA2R-Ab)(32.92±6.19) RU/mL vs (9.37±1.92) RU/mL, 24h urine protein quantification(7.96±0.94) g/d vs (0.11±0.02) g/d, lncRNA GAS5-AS1(3.46±0.56) vs (1.02±0.17), IL-27(124.76±20.77) ng/L vs (62.71±10.44) ng/L, Th17(4.86±0.81)% vs (0.75±0.12)%, Th17/Treg(1.43±0.23) vs (0.11±0.02) in PMN group increased (P<0.05), while glomerular filtration rate(GFR)(57.47±10.24) mL·min⁻¹·（1.73 m²）⁻¹ vs (109.22±14.85) mL·min⁻¹·（1.73 m²）⁻¹ and Treg levels(3.21±0.53)% vs (6.42±1.07)% decreased (P<0.05); Comparison of Th17/Treg, IL-17, TNF-α, IL-10, TGF-β1, lncRNA GAS5-AS1, and IL-27 levels in patients with PMN at different levels of risk, and the differences were statistically significant(P<0.05); the peripheral blood lncRNA GAS5-AS1 of PMN patients was positively correlated with Th17/Treg, L-17, and TNF-α(r=0.632, 0.554, 0.572, P<0.05), and negatively correlated with IL-10(r =−0.468, P<0.05); IL-27 was positively correlated with Th17/Treg and IL-17(r=0.581, 0.602, P<0.05), and negatively correlated with IL-10 and TGF-β1(r=−0.533, −0.436, P<0.05).

Conclusion  Serum lncRNA GAS5-AS1 and IL-27 are highly expressed in PMN patients, both of them regulate Th17/Treg immune balance, play an important role in PMN, and are expected to become the research targets of PMN.