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摘要：
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摘要：目的 探讨代谢相关脂肪性肝病（metabolic associated fatty liver disease，
MAFLD）中微小RNA（microRNA，miRNA）对铁死亡的调控。方法 在GSE135251
数据集中进行差异表达分析及基因集变异分析（gene set variation analysis，GSVA）。
在GSE114923数据集中进行差异分析，并识别靶向调控铁死亡相关基因的miRNA。以
2022年3月1日至2023年4月30日于新疆医科大学第一附属医院经腹部超声确诊为MAFLD的
10例患者作为MAFLD组，以同期性别和年龄匹配的10例健康志愿者作为健康对照组，
采集外周血样本。通过反转录定量聚合酶链式反应（reverse transcription quantitative
polymerase chain reaction，RT-qPCR）和Western blot检测miRNA及铁死亡相关蛋白
[（溶质载体家族7成员11（solute carrier family 7 member 11，SLC7A11）、谷胱甘肽过
氧化物酶4（glutathione peroxidase 4，GPX4）、血红素加氧酶1（heme oxygenase 1，
HMOX1）] 的表达。通过酶联免疫吸附试验检测谷胱甘肽（glutathione，GSH）、丙二
醛（malondialdehyde，MDA）和炎症因子 [白细胞介素6（interleukin-6，IL-6）、肿瘤
坏死因子α（tumor necrosis factor α，TNF-α）] 水平。结果 GSE135251和GSE114923数
据集分析表明，与健康对照组相比，MAFLD患者中细胞坏死（t = 3.229，P = 0.022）
和铁死亡（t = 2.008，P = 0.006）通路显著激活，差异分析结果表明MAFLD和对照
组间有8687个有差异表达的基因，其中有31个为铁死亡相关基因。在31个差异表达的
miRNA中鉴定了7个靶向调控铁死亡的miRNA，hsa-miR-192-5p（4.628 ± 1.234比3.171 ± 0.456；
t = 2.217，P = 0.068）、hsa-miR-122-5p（13.532 ± 0.946比10.536 ± 1.444；t = 3.472，
P = 0.013）、hsa-miR-30a-5p（6.081 ± 0.770比4.106 ± 0.269；t = 4.841，P = 0.003）、
hsa-miR-100-5p（5.888 ± 0.933比3.888 ± 0.721；t = 3.395，P = 0.015）在MAFLD患者
中表达上调，hsa-miR-10b-5p（5.077 ± 0.876比6.439 ± 1.076；t = 1.963，P = 0.097）、
hsa-miR-223-5（0.626 ± 0.723比2.790 ± 1.912；t = 2.111，P = 0.079）、hsa-miR-215-
5p（0.595 ± 0.771比2.738 ± 0.885，t = 3.652，P = 0.011）表达下调，其中hsa-miR-30a-
5p上调水平最显著。RT-qPCR结果表明，与对照组比较，MAFLD患者中hsa-miR-30a-
5p表达水平显著升高（1.591 ± 0.229比1.012 ± 0.031；t = 9.676，P ＜ 0.001），SLC7A11
mRNA（0.598 ± 0.108比0.997 ± 0.034；t = 13.64，P ＜ 0.001）、GPX4 mRNA（0.724 ±
0.064比1.003 ± 0.029，t = 15.34；P ＜ 0.001）和HMOX1 mRNA（0.688 ± 0.078比0.993 ±
0.034；t = 13.92，P ＜ 0.001）表达水平显著降低。Western blot结果显示，与对照组
相比，SLC7A11蛋白（0.712 ± 0.074比1.000 ± 0.053；t = 10.01，P ＜ 0.001）、GPX4
蛋白（0.810 ± 0.034比1.000 ± 0.019；t = 15.25，P ＜ 0.001）和HMOX1蛋白（0.673 ±
0.026比1.000 ± 0.029；t = 26.75，P ＜ 0.001）在MAFLD患者中表达显著下调。与健康
对照组相比，MAFLD患者GSH [（163.684 ± 15.857） U/g比（197.728 ± 11.009）U/g；t =
6.109，P ＜ 0.001] 水平显著降低，MDA [（2.494 ± 0.253）μmol/g比（1.0612 ± .205）μmol/g；
t = 7.602，P = 0.002]、IL-6 [（55.219 ± 0.743）ng/L比（46.456 ± 1.831）ng/L；t =
14.48，P ＜ 0.001] 和TNF-α [（22.883 ± 2.893）μg/L比（13.885 ± 0.169）μg/L；t =
10.78，P ＜ 0.001] 水平显著升高。结论 hsa-miR-30a-5p可能通过靶向SLC7A11促进铁
死亡，在MAFLD中发挥促炎和促氧化作用。
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Abstract: Objective To investigate the regulation of ferroptosis by microRNA (miRNA) in
metabolic associated fatty liver disease (MAFLD). Methods Differential expression analysis
and gene set variation analysis (GSVA) were conducted in GSE135251 data set. Subsequently,
differential analysis was performed in GSE114923 data set, and the regulation of miRNA
targeting ferroptosis-related genes were identified. Ten patients diagnosed with MAFLD by
abdominal ultrasound in the First Affiliated Hospital of Xinjiang Medical University from
March 1st, 2022 to April 30th 2023 were selected as the MAFLD group, and ten healthy
volunteers matched with gender and age in the same period were selected as the health control
group. Peripheral blood samples were collected. The expression of miRNA and ferroptosisrelated
proteins [solute carrier family 7 member 11 (SLC7A11), glutathione peroxidase 4
(GPX4), heme oxygenase 1 (HMOX1) ] were detected by reverse transcription quantitative
polymerase chain reaction (RT-qPCR) or Western blot. The levels of glutathione (GSH),
malondialdehyde (MDA) and inflammatory factors [interleukin-6 (IL-6), tumor necrosis factor
α (TNF-α)] were measured by enzyme-linked immunosorbent assay. Results Analysis of
the GSE135251 and GSE114923 data sets showed that compared with the control group, the
cell necrosis (t = 3.229, P = 0.022) and ferroptosis (t = 2.008, P = 0.006) pathways were
significantly activated in MAFLD patients. Furtherly, 8687 differentially expressed genes
were identified, among which 31 were ferroptosis-related genes. Among 31 differentially
expressed miRNA, 7 miRNA were identified to regulate ferroptosis, of which hsa-miR-192-5p
(4.628 ± 1.234 vs. 3.171 ± 0.456; t = 2.217, P = 0.068), hsa-miR-122-5p (13.532 ± 0.946 vs.
10.536 ± 1.444; t = 3.472, P = 0.013), hsa-miR-30a-5p (6.081 ± 0.770 vs. 4.106 ± 0.269; t = 4.841,
P = 0.003), hsa-miR-100-5p (5.888 ± 0.933 vs. 3.888 ± 0.721; t = 3.395, P = 0.015) were
upregulated in MAFLD patients, while hsa-miR-10b-5p (5.077 ± 0.876 vs. 6.439 ± 1.076; t =
1.963, P = 0.097), hsa-miR-223-5 (0.626 ± 0.723 vs. 2.790 ± 1.912; t = 2.111, P = 0.079), hsamiR-
215-5p (0.595 ± 0.771 vs. 2.738 ± 0.885; t = 3.652, P = 0.011) were downregulated. The
upregulation level of hsa-miR-30a-5p was the most significant. RT-qPCR results showed that
compared with the control group, the expression level of hsa-miR-30a-5p (1.591 ± 0.229 vs.
1.012 ± 0.031, t = 9.676, P ＜ 0.001) was significantly increased in MAFLD patients, while the
expression levels of SLC7A11 mRNA (0.598 ± 0.108 vs. 0.997 ± 0.034; t = 13.64, P ＜ 0.001),
GPX4 mRNA (0.724 ± 0.064 vs. 1.003 ± 0.029, t = 15.34; P ＜ 0.001) and HMOX1 mRNA
(0.688 ± 0.078 vs. 0.993 ± 0.034; t = 13.92, P ＜ 0.001) were significantly decreased. Western
blot showed that compared with the control group, SLC7A11 protein (0.712 ± 0.074 vs. 1.000 ±
0.053; t = 10.01, P ＜ 0.001), GPX4 protein (0.810 ± 0.034 vs. 1.000 ± 0.019; t = 15.25, P ＜
0.001), and HMOX1 protein (0.673 ± 0.026 vs. 1.000 ± 0.029; t = 26.75, P ＜ 0.001) were
significantly downregulated in MAFLD patients. Compared with the control group, GSH level
[(163.684 ± 15.857) U/g vs. (197.728 ± 11.009) U/g; t = 6.109, P ＜ 0.001] in MAFLD group
decreased significantly, MDA [(2.494 ± 0.253) μmol/g vs. (1.0612 ± 0.205) μmol/g; t = 7.602,
P = 0.002], IL-6 [(55.219 ± 0.743) ng/L vs. (46.456 ± 1.831) ng/L; t = 14.48, P ＜ 0.001] and
TNF-α [(0.022 ± 0.002) μg/L vs. (0.013 ± 0.001) μg/L; t = 10.78, P ＜ 0.001] levels increased
significantly. Conclusion Hsa-miR-30a-5p may promote ferroptosis levels by targeting
SLC7A11, playing a pro-inflammatory and prooxidant role in MAFLD.
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