原发性膜性肾病患者外周血lncRNA GAS5-AS1、白细胞介素-27表达及与Treg/Th17平衡的关系

钟娇影; 刘杰; 刘慧

doi:10.3969/j.issn.1671-2390.2023.12.004

原发性膜性肾病患者外周血lncRNA GAS5-AS1、白细胞介素-27表达及与Treg/Th17平衡的关系

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

摘要

摘要:
目的检测原发性膜性肾病（primary membranous nephropathy，PMN）患者外周血长链非编码RNA-生长停滞特异性转录本5反义RNA1（long non-coding RNA growth arrest-specific transcript 5 antisense RNA1，lncRNA GAS5-AS1）、白细胞介素（interleukin，IL）-27表达，并分析其与患者调节性T细胞/辅助性T细胞17（regulatory T cell/T help 17cells，Treg/Th17）平衡的关系。
方法选择2019年9月至2020年9月间河北以岭医院收治的PMN患者94例作为PMN组。另选取同期体检健康者96例作为对照组。收集所有研究对象外周血标本，实时荧光定量PCR（real-time quantitative PCR，qRT-PCR）法测定血清lncRNA GAS5-AS1水平、酶联免疫吸附法（enzyme-linked immunosorbent assay，ELISA）法测定IL-27、IL-17、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、IL-10、转化生长因子β1（transforming growth factor betal，TGF-β1）水平，流式细胞仪测定外周血Treg细胞、Th17细胞比例，并计算Treg/Th17比值。Pearson法分析PMN患者lncRNA GAS5-AS1、IL-27水平与IL-17、TNF-α、IL-10、TGF-β1、Treg/Th17的相关性。
结果对照组血尿素氮（blood urea nitrogen，BUN）为（4.03 ± 0.66）mmol/L，PMN组患者BUN为（6.52 ± 1.05）mmol/L；对照组血尿酸（uric acid，UA）为（266.13 ± 44.35）µmol/L，PMN组患者UA为（338.92 ± 56.45）µmol/L；对照组胱抑素C（cystatin C，Cys C）为（0.72 ± 0.12）mg/L，PMN组患者Cys C 为（1.38 ± 0.23）mg/L；对照组抗磷脂酶A2受体抗体（phospholipase A2 receptor antibody，PLA2R-Ab）为（9.37 ± 1.92）RU/mL，PMN组患者PLA2R-Ab为（32.92 ± 6.19）RU/mL；对照组24 h尿蛋白定量（0.11 ± 0.02）g，PMN组患者24 h尿蛋白定量（7.96 ± 0.94）g；对照组lncRNA GAS5-AS1为（1.02 ± 0.17），PMN组患者lncRNA GAS5-AS1 为（3.46 ± 0.56）；对照组IL-27为（62.71 ± 10.44）ng/L，PMN组患者IL-27为（124.76 ± 20.77）ng/L；对照组Th17为（0.75 ± 0.12）%，PMN组患者Th17为（4.86 ± 0.81）%；对照组Th17/Treg为（0.11 ± 0.02），PMN组患者Th17/Treg为（1.43 ± 0.23）。对照组肾小球滤过率（glomerular filtration rate，GFR）为（109.22 ± 14.85）mL·min⁻¹·（1.73 m²）⁻¹，PMN组患者GFR为（57.47 ± 10.24）mL·min⁻¹·（1.73 m²）⁻¹；对照组Treg水平为（6.42 ± 1.07）%，PMN组患者Treg水平为（3.21 ± 0.53）%。不同危险程度PMN患者Th17/Treg、IL-17、TNF-α、IL-10、TGF-β1、lncRNA GAS5-AS1、IL-27水平差异均有统计学意义（P＜0.05）；PMN患者外周血lncRNA GAS5-AS1与Th17/Treg、IL-17、TNF-α正相关（r = 0.632、0.554、0.572，P＜0.05），与IL-10负相关（r = −0.468，P＜0.05）；IL-27与Th17/Treg、IL-17正相关（r = 0.581、0.602，P＜0.05），与IL-10、TGF-β1负相关（r = −0.533、−0.436，P＜0.05）。
结论 PMN患者血清lncRNA GAS5-AS1、IL-27均高表达，二者均调节Th17/Treg免疫平衡，在PMN中发挥重要作用，有望成为PMN的研究靶点。

Abstract:
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.

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