The relationship between peripheral blood lncRNA H19 expression and Th17/Treg balance in patients with acute pancreatitis
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摘要: 目的 探究急性胰腺炎(AP)患者外周血长链非编码RNA H19(lncRNA H19)表达与辅助性T细胞17(Th17)/调节性T细胞(Treg)平衡的关系。方法 以2019年9月—2021年9月收治的91例AP患者为研究对象，根据患者病情程度分为轻度急性胰腺炎(MAP)34例，中度急性胰腺炎(MSAP)31例，重度急性胰腺炎(SAP)26例；另选取75例健康体检者为对照组。各组外周血lncRNA H19水平检测采用实时荧光定量PCR(qRT-PCR)法；Th17、Treg细胞水平采用流式细胞术检测；外周血IL-17、IL-6、IL-10、转化生长因子-β(TGF-β)水平利用酶联免疫吸附法检测。Pearson相关性分析外周血lncRNA H19与Th17/Treg及其相关细胞因子的关系。结果 各组之间检测降钙素原、C反应蛋白、总胆固醇、三酰甘油、高密度脂蛋白、低密度脂蛋白水平、急性生理学和慢性健康状况评分比较均差异有统计学意义(P>0.05)。与对照组相比，AP组患者外周血lncRNA H19、Th17细胞比例、Th17/Treg比值、IL-17、IL-6水平均明显升高，且随疾病进展而显著上升，而Treg细胞比例、IL-10、TGF-β水平明显下降，且随疾病加重而显著下降(P< 0.05)。Pearson相关性分析结果显示，AP组患者外周血lncRNA H19水平与Th17、Th17/Treg、IL-17、IL-6呈显著正相关(P< 0.05)，与Treg、IL-10、TGF-β呈显著负相关(P< 0.05)。结论 AP患者外周血lncRNA H19表达水平及Th17/Treg比值均增加，其机制可能是lncRNA H19通过影响Th17/Treg平衡而参与AP的发生、发展。
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关键词:
- 急性胰腺炎 /
- 长链非编码RNA H19 /
- 辅助性T细胞17 /
- 调节性T细胞
Abstract: Objective To explore the relationship between the expression of long non-coding RNA H19(lncRNA H19) in peripheral blood of patients with acute pancreatitis(AP) and the balance of helper T cell 17(Th17)/regulatory T cell(Treg).Methods Ninety-one patients with AP who accepted by our hospital from September 2019 to September 2021 were gathered as the research objects. According to the severity of the patients, they were separated into 34 cases of mild acute pancreatitis(MAP), 31 cases of moderate acute pancreatitis(MSAP) and 26 cases of severe acute pancreatitis(SAP), and 75 healthy subjects were gathered as control group. The level of lncRNA H19 in peripheral blood was measured by real-time quantitative PCR(qRT-PCR) method; the levels of Th17 and Treg cells in each group were measured by flow cytometry; the levels of interleukin-17(IL-17), interleukin-6(IL-6), interleukin-10(IL-10) and transforming growth factor-β(TGF-β) in peripheral blood were measured by enzyme-linked immunosorbent assay(ELISA). The relationship between peripheral blood lncRNA H19 and Th17/Treg and its related cytokines was analyzed by Pearson correlation analysis.Results There were significant differences in PCT, CRP, TC, TG, HDL-C, LDL-C, and APACHEⅡ among patients in each group(P>0.05). Peripheral blood lncRNA H19, Th17 cell ratio, Th17/Treg ratio, IL-17, IL-6 levels in AP patients increased greatly than control group, and increased greatly with disease progression, while Treg cell ratio, IL-10, TGF-β levels decreased greatly, and decreased greatly with the aggravation of the disease(P< 0.05). The results of Pearson correlation analysis showed that the level of lncRNA H19 in peripheral blood of AP patients was greatly directly correlated with Th17, Th17/Treg, IL-17, and IL-6(P< 0.05), and was greatly inversely correlated with Treg, IL-10, and TGF-β(P< 0.05).Conclusion The expression level of lncRNA H19 and the ratio of Th17/Treg in peripheral blood of AP patients are increased, and the mechanism may be that lncRNA H19 participates in the occurrence and development of AP by affecting the Th17/Treg balance.-
Key words:
- acute pancreatitis /
- long non-coding RNA H19 /
- helper T cell 17 /
- regulatory T cell
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表 1 qRT-PCR引物序列
基因 正向引物5’-3’ 反向引物5’-3’ lncRNA H19 TTCAAAGCCTCCACGACTCT GCTCACACTCACGCACACTC GAPDH CGGAGTCAACGGATTTGGTCGTAT AGCCTTCTCCATGGTGGTGAAGAC 
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表 2 各组受试者临床资料比较
例(%)，X±S 临床资料 对照组(n=75) MAP组(n=34) MSAP组(n=31) SAP组(n=26) F/χ2 P 年龄/岁 42.24±5.11 43.15±5.15 43.42±5.17 43.73±5.19 0.769 0.513 性别 男 41(54.66) 18(52.94) 17(54.83) 15(57.69) 0.136 0.987 女 34(45.34) 16(47.06) 14(45.17) 11(42.31) BMI 23.69±3.63 24.02±3.69 24.47±3.75 24.68±3.81 0.624 0.600 高血压 5(6.67) 3(8.82) 4(12.90) 4(15.38) 2.132 0.545 糖尿病 3(4.00) 2(5.88) 3(9.68) 3(11.54) 2.251 0.522 冠心病 4(5.33) 3(8.82) 4(12.90) 4(15.38) 3.028 0.387 PCT/(μg·L-1) 0.37±0.09 0.68±0.111) 1.21±0.241)2) 1.73±0.311)2)3) 442.740 < 0.001 CRP/(mg·L-1) 3.92±0.83 9.14±2.051) 14.08±3.881)2) 17.19±4.471)2)3) 212.055 < 0.001 TC/(mmol·L-1) 3.32±1.05 4.21±1.251) 5.19±1.381)2) 5.87±1.541)2)3) 34.738 < 0.001 TG/(mmol·L-1) 2.66±0.68 3.24±0.811) 4.31±1.031)2) 5.66±1.421)2)3) 76.765 < 0.001 HDL-C/(mmol·L-1) 1.16±0.24 1.02±0.201) 0.87±0.161)2) 0.72±0.141)2)3) 35.403 < 0.001 LDL-C/(mmol·L-1) 1.94±0.52 2.85±0.781) 3.71±1.111)2) 3.94±1.191)2)3) 55.160 < 0.001 APACHEⅡ评分/分 10.23±2.07 18.33±2.682) 26.04±4.372)3) 195.903 < 0.001 与对照组相比，1)P < 0.05；与MAP组相比，2)P < 0.05；与MSAP组相比，3)P < 0.05。
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表 3 AP组与对照组lncRNA H19及Th17、Treg细胞水平比较
X±S 组别 例数 lncRNA H19/GAPDH Th17/% Treg/% Th17/Treg 对照组 75 1.02±0.17 2.71±0.35 10.65±1.83 0.25±0.06 AP组 91 2.10±0.32 4.40±0.49 7.03±0.95 0.63±0.12 t 26.318 25.057 16.387 24.964 P < 0.001 < 0.001 < 0.001 < 0.001
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表 4 不同严重程度AP患者lncRNA H19及Th17、Treg细胞水平比较
X±S 组别 例数 lncRNA H19/GAPDH Th17/% Treg/% Th17/Treg MAP组 34 1.63±0.22 3.51±0.42 9.04±1.31 0.39±0.08 MSAP组 31 2.11±0.311) 4.63±0.501) 6.64±0.871) 0.67±0.111) SAP组 26 2.72±0.431)2) 5.28±0.571)2) 4.86±0.591)2) 0.89±0.181)2) F 84.619 99.937 132.194 119.689 P < 0.001 < 0.001 < 0.001 < 0.001 与MAP组相比，1)P < 0.05；与MSAP组相比，2)P < 0.05。
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表 5 AP组与对照组细胞因子水平比较
pg/mL，X±S 组别 例数 IL-17 IL-6 IL-10 TGF-β 对照组 75 17.65±3.82 72.68±15.27 67.19±8.11 31.42±5.07 AP组 91 40.26±6.32 120.57±19.31 42.94±6.44 14.41±3.26 t 27.155 17.445 21.473 26.124 P < 0.001 < 0.001 < 0.001 < 0.001
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表 6 不同严重程度AP患者细胞因子水平比较
pg/mL，X±S 组别 例数 IL-17 IL-6 IL-10 TGF-β MAP组 34 29.14±4.81 94.19±17.31 50.56±7.24 22.62±4.23 MSAP组 31 41.17±6.941) 127.94±18.771) 42.16±6.351) 11.49±3.281) SAP组 26 53.74±7.621)2) 146.28±22.341)2) 33.92±5.511)2) 7.17±1.961)2) F 107.648 56.821 48.877 170.851 P < 0.001 < 0.001 < 0.001 < 0.001 与MAP组相比，1)P < 0.05；与MSAP组相比，2)P < 0.05。
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表 7 AP患者外周血lncRNA H19与Th17/Treg及其相关细胞因子的关系
指标 lncRNA H19 r P Th17 0.397 0.005 Treg -0.376 0.007 Th17/Treg 0.489 < 0.001 IL-17 0.387 0.006 IL-6 0.341 0.009 IL-10 -0.349 0.008 TGF-β -0.372 0.007
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