miR-26a-5p通过JAK2/STAT3 信号通路对人类风湿关节炎成纤维样滑膜细胞凋亡的影响

doi:10.13481/j.1671-587X.20210227

Abstract

Abstract: Objective

To explore the effects of miR-26a-5p on the apoptosis and secretion of inflammatory factors in the rheumatoid arthritis （RA）-fibroblast-like synovial cells （FLSs）， and to clarify its mechanism of action.

Methods

The RA-FLSs from knee joint synovial tissue of patients with RA were isolated and cultured，and miR-26a-5p mimics， inhibitors and negative controls were transfected into the RA-FLSs.Real-time fluorescence quantitative polymerase chain reaction（qRT-PCR） was used to detect the effect of transfection， ELISA was used to detect the levels of interleukin-1β （IL-1β）， interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α） in cell culture fluid， CCK-8 method was used to detect the cell proliferation activity， AnnexinⅤ-FITC/PI and Hoechst 33342 staining were used to observe the apoptosis， and Western blotting method was used to detect the expression levels of apoptosis-related proteins B cell lymphoma-2（Bcl-2），Bcl-2 associated X protein（Bax） and Janus kinase 2（JAK2）/signal transducer and activator of transcription（STAT3） signaling pathway-related proteins in the cells.

Results

Compared with blank control group and NC-mimic group， the expression level of miR-26a-5p in RA-FLSs in miR-26a-5p mimic group was increased （P<0.05）， the proliferation activity of RA-FLSs was decreased（P<0.05），the apoptotic rate of cells was increased （P<0.05），the expression levels of Bcl-2， JAK2， p-JAK2 and STAT3 proteins were decreased（P<0.05）， and the expression level of Bax protein was increased （P<0.05）.Compared with blank control group and NC-inhibitor group， the expression level of miR-26a-5p in RA-FLSs in miR-26a-5p inhibitor group was decreased （P<0.05）， the proliferation activity of RA-FLSs was increased （P<0.05）， the apoptotic rate was decreased （P<0.05）， the expression levels of Bcl-2，JAK2， p-JAK2 and STAT3 proteins were increased （P<0.05）， and the expression level of Bax protein was decreased （P<0.05）.

Conclusion

Overexpression of miR-26a-5p can inhibit the proliferation of RA-FLSs and promote apoptosis. Its mechanism may be related to the inhibition of the activation of JAK2/STAT3 signaling pathway.

Key words: miR-26a-5p, rheumatoid arthritis, fibroblast-like synoviocyte, apoptosis, inflammatory factor

CLC Number:

R593.22

Jie WU,Xuehua YANG,Ling MA,Wenqiang FAN,Dongdong FU,Xiao GAO,Shufei ZUO,Shu LIANG,Yilu QIN,Peishan WANG,Jinyan GUO. Effect of miR-26a-5p on apoptosis of human rheumatoid arthritis fibroblast-like synovial cells through JAK2/STAT3 signaling pathway[J].Journal of Jilin University(Medicine Edition), 2021, 47(2): 460-468.

Figures/Tables 8

Fig. 1

Tab.1

Tab.2

Fig. 2

Tab.3

Fig. 3

Fig.4

Tab.4

References 21

1	SONG L， YIN Q， KANG M， et al. Untargeted metabolomics reveals novel serum biomarker of renal damage in rheumatoid arthritis ［J］. J Pharm Biomed Anal，2020，180： 113068.
2	IZQUIERDO E， CAÑETE J D， CELIS R， et al. Synovial fibroblast hyperplasia in rheumatoid arthritis： clinicopathologic correlations and partial reversal by anti-tumor necrosis factor therapy ［J］. Arthritis Rheum， 2011， 63（9）： 2575-2583.
3	ALAM J， JANTAN I， BUKHARI S N A. Rheumatoid arthritis： recent advances on its etiology， role of cytokines and pharmacotherapy ［J］. Biomed Pharmacother， 2017， 92： 615-633.
4	张阳，王永萍，潘书涵. 类风湿关节炎滑膜细胞与自噬研究进展［J］. 贵阳中医学院学报， 2019， 41（5）：97-100.
5	MCINNES I B， SCHETT G. Pathogenetic insights from the treatment of rheumatoid arthritis ［J］. Lancet， 2017， 389（10086）： 2328-2337.
6	XU J F， YANG G H， PAN X H， et al. Altered microRNA expression profile in exo-somes during osteogenic differentiation of human bone marrow-derived mesenchymal stem cells［J］.PLoS One，2014，9（12）： e114627.
7	RASHEED Z， AL-SHOBAILI H A， RASHEED N， et al. MicroRNA-26a-5p regulates the expression of inducible nitric oxide synthase via activation of NF-κB pathway in human osteoarthritis chondrocytes ［J］.Arch Biochem Biophys， 2016， 594： 61-67.
8	ZHANG L， LIU X， CHE S， et al. CircRNA-9119 regulates the expression of prostaglandin-endoperoxide synthase 2 （PTGS2） by sponging miR-26a in the endometrial epithelial cells of dairy goat ［J］. Reprod Fertil Dev， 2018， 30（12）： 1759-1769.
9	SUN J， FENG M， WU F， et al. Plasma miR-26a as a diagnostic biomarker regulates cytokine expression in systemic juvenile idiopathic arthritis［J］. J Rheumatol， 2016， 43（8）：1607-1614.
10	TAO W， DONG X， KONG G，et al. Elevated circulating hsa-miR-106b， hsa-miR-26a， and hsa-miR-29b in type 2 diabetes mellitus with diarrheaPredominant irritable bowel syndrome［J］. Gastroenterol Res Pract， 2016， 2016： 9256209.
11	郭雄飞，王挺，汤立新. miR-29a对类风湿关节炎成纤维样滑膜细胞增殖和凋亡的影响［J］.基础医学与临床， 2020， 40（1）：9-15.
12	庞雅湘，韩华，殷玥，等. 过表达miR-140-5p促进前列腺癌细胞凋亡抑制其增殖［J］.河北医科大学学报， 2020， 41（1）：7-11.
13	GAO J， ZHOU X， KONG R， et al. microRNA-126 targeting PIK3R2 promotes rheumatoid arthritis synovial fibro-blasts proliferation and resistance to apoptosis by regulating PI3K/AKT pathway［J］. Exp Mol Pathol， 2016， 100 （1）：192-198.
14	PHILIPPE L， ALSALEH G， SUFFERT G， et al. TLR2 expression is regulated by MicroRNA miR-19 in rheumatoid fibroblast-like synoviocytes［J］. J Immunol， 2012， 188 （1）： 454-461.
15	BOTTINI N， FIRESTEIN G S. Duality of fibroblast-like synoviocytes in RA： passive responders and imprinted aggressors［J］.Nat Rev Rheumatol，2013，9（1）： 24-33.
16	TERENZI R， ROMANO E， MANETTI M， et al. Neuropeptides activate TRPV1 in rheumatoid arthritis fibroblast-like synoviocytes and foster IL-6 and IL-8 production［J］. Ann Rheum Dis， 2013， 72（6）：1107-1109.
17	蒲金呈，王璇，汤建平.长链非编码RNA在风湿免疫病发病分子机制中的研究进展［J］.同济大学学报（医学版），2020，41（6）：789-795.
18	戴小娜，林进.老年起病类风湿关节炎研究进展［J］.中国实用内科杂志，2019，39（6）：562-567.
19	江国强，方芳. 厄洛替尼联合康莱特对肺癌A549细胞增殖、侵袭及JAK2/STAT3信号通路的影响［J］.郑州大学学报（医学版）， 2019， 54（3）：418-422.
20	LOU L， LIU Y， ZHOU J， et al. Chlorogenic acid and luteolin synergistically inhibit the proliferation of interleukin-1beta-induced fibroblast-like synoviocytes through regulating the activation of NF-kappaB and JAK/STAT-signaling pathways［J］. Immunopharmacol Immunotoxicol， 2015， 37 （6）： 499-507.
21	YANG Q J， JIN-DI Y D， PAN D S， et al. Advance in research of selective JAK inhibitors for treating rheumatoid arthritis［J］.Chin J New Drugs，2015，24（1）：39-45.

Group	miR-26a-5p	IL-1β[ρ_B/(ng·L^-¹)]	IL-6[ρ_B/ (ng·L^-¹)]	TNF-α[ρ_B/ (ng·L^-¹)]
Blank control	1.00±0.00	15.53±1.64	92.21±4.17	274.56±26.39
NC-mimic	0.99±0.01	15.36±1.39	92.18±3.97	273.12±27.08
miR-26a-5p mimic	3.84±0.24^*△	11.87±1.05^*△	72.56±3.45^*△	228.25±19.23^*△
NC-inhibitor	1.01±0.02	15.45±1.73	91.87±4.52	273.87±24.62
miR-26a-5p inhibitor	0.34±0.00^*#	19.16±2.01^*^#	122.49±7.23^*^#	298.56±29.32^*^#
F	92.097	132.724	87.193	72.372
P	<0.01	<0.01	<0.01	<0.01

Group	Proliferation activity
Group	(t/h) 0	12	24	48	72
Blank control	0.24±0.01	0.30±0.02	0.52±0.04	0.64±0.05	0.81±0.05
NC-mimic	0.23±0.02	0.30±0.01	0.51±0.03	0.63±0.04	0.80±0.07
miR-26a-5p mimic	0.23±0.00	0.29±0.01	0.31±0.04^*△	0.38±0.02^*△	0.42±0.03^*△
NC-inhibitor	0.24±0.02	0.29±0.00	0.52±0.03	0.64±0.05	0.80±0.06
miR-26a-5p inhibitor	0.23±0.01	0.30±0.02	0.69±0.04^*^#	0.81±0.06^*^#	0.98±0.10^*^#
F	0.203	0.117	34.472	50.452	84.324
P	0.912	0.936	<0.01	<0.01	<0.01

Group	Apoptotic rate
Blank control	7.57±0.82
NC-mimic	7.72±0.76
miR-26a-5p mimic	21.41±1.95^*△
NC-inhibitor	8.69±0.76
miR-26a-5p inhibitor	4.20±0.32^*^#
F	104.234
P	<0.01

Group	Bcl-2	Bax	JAK2	p-JAK2	STAT3	p-STAT3
Blank control	0.76±0.05	0.37±0.01	0.51±0.03	1.15±0.04	0.79±0.06	0.45±0.03
NC-mimic	0.77±0.06	0.38±0.02	0.52±0.05	1.19±0.07	0.80±0.05	0.46±0.03
miR-26a-5p mimic	0.14±0.00^*△	0.72±0.05^*△	0.21±0.00^*△	0.79±0.05^*△	0.20±0.01^*△	0.17±0.00^*△
NC-inhibitor	0.77±0.04	0.37±0.02	0.52±0.04	1.20±0.11	0.79±0.02	0.45±0.02
miR-26a-5p inhibitor	2.01±0.13^*^#	0.13±0.01^*^#	1.36±0.08^*^#	2.24±0.17^*^#	1.56±0.04^*^#	0.70±0.05^*^#
F	81.281	68.027	120.338	79.092	87.877	56.302
P	<0.01	<0.01	<0.01	<0.01	<0.01	<0.01

[1]	Yang LI,Shiyang LI,Hongmei ZHANG,Qianqian LIU,Yubing CHEN. Effect of NGFR gene expression differences on promotion of apoptosis of squamous cell carcinoma and adenocarcinoma cells by PD-1 inhibitor-activated T lymphocytes [J]. Journal of Jilin University(Medicine Edition), 2026, 52(3): 680-693.
[2]	Qiaotong REN,Weihua WU,Xingxiang WANG,Shichao WANG,Yipeng CHENG,Chun LI. Effect of interleukin-17 recepter B gene silencing on proliferation, invasion and apoptosis of lung adenocarcinoma A549 cells and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2026, 52(3): 738-745.
[3]	Qiuping SU,Lei XING,Wei ZHANG,Shan JIANG,Yangyang ZHAO,Fangfang CHEN. Research progress in anti-tumor mechanism of berberine and application of its novel nano-delivery system in tumor therapy [J]. Journal of Jilin University(Medicine Edition), 2026, 52(3): 863-871.
[4]	Jie YU,Zongkang WANG,Xun LIU,Yanan YANG,Jin TAN. Effect of miR-153-3p/PGC-1α axis on apoptosis of oral squamous cell carcinoma cells by modulation of mitochondrial apoptosis pathway [J]. Journal of Jilin University(Medicine Edition), 2026, 52(2): 348-358.
[5]	Yaping LI,Chunyan TAN,Lujie ZHAO,Jiayi ZHAO,Ting LI,Xiao YANG,Xiaoyun YANG. Inductive effect of lysophosphatidic acid combined with 6-hydroxydopamine on apoptosis of SH-SY5Y cells and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2026, 52(1): 152-161.
[6]	Jiaxin WANG,Junwen MAO,Xuan ZHANG. Expression of natural autoantibodies against apoptosis inhibitory genes in plasma of patients with hepatocellular carcinoma and its clinical significance [J]. Journal of Jilin University(Medicine Edition), 2026, 52(1): 228-235.
[7]	Huaimin LIANG,Jiacheng JIN,Wenhua CHEN,Yuyao LI,Hangyu WANG,Ke ZHANG,Jinhui WANG. UPLC-Q-TOF/MS and network pharmacology analysis and experimental verification based on potential active ingredients and mechanisms of medicinal Mulberry Leaves in anti-acute kidney injury [J]. Journal of Jilin University(Medicine Edition), 2026, 52(1): 56-69.
[8]	Shanshan SUN,Mei LU,Xinfu GAO,LYu Guangyao,Baolei Zhao,LYu Wenwen. Inhibitory effect of Bradykinin 1 receptor antagonist ELN441958 on proliferation of HepG2 cells by regulating Akt/FoxO3a signaling pathway [J]. Journal of Jilin University(Medicine Edition), 2026, 52(1): 70-80.
[9]	Xiaohan YAO,Mingchen YAO,Zhiqing WANG,Heyang LI,Yan YAN,Ningjing LEI. Effect of silencing GPR139 gene on proliferation, apoptosis and autophagy of breast cancer cells and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2026, 52(1): 1-9.
[10]	Han XUE,Yuxin FAN,Ting ZHANG,Zhimin LI,Mingge HUO,Xingang GUAN. Preparation of nanodrug PTX₂ NPs and its killing effect on human lung cancer A549 cells [J]. Journal of Jilin University(Medicine Edition), 2025, 51(5): 1260-1266.
[11]	Ying HU,Yong HUANG. Improvement effect of short-chain fatty acids on inflammation-induced autophagic damage in ovarian granulosa cells in polycystic ovary syndrome and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2025, 51(5): 1340-1348.
[12]	Xing YUE,Xuemei LI,Hanxiao ZHANG,Chuanyi ZUO,Lijuan ZHU,Jing LYU,Chengshun ZHANG,Xin CAO. Role of serum total bile acid level in development of arrhythmia in ApoE^-/- mice [J]. Journal of Jilin University(Medicine Edition), 2025, 51(4): 879-886.
[13]	Jiarui LI,Zhenlin YANG,Fan GAO,Jingjing GUO,Jinzi LI. Effect of miR-34a-5p on hippocampal neuron apoptosis in rats with temporal lobe epilepsy and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2025, 51(4): 939-947.
[14]	Panxi SUN,Xue QIN,Chongyang ZHANG,Jia LUO,Yong CHEN,Lili WEI. Protective effect of TUG-891 on ischemic stroke induced by ischemia and hypoxia and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2025, 51(4): 968-975.
[15]	Cheng CHEN,Jingyao LI,Wanxiang HU,Donghui LIU,Zhihong CHEN. Protective effect of sericin on streptozotocin-induced INS-1 cell damage by regulating PI3K/Akt/NF-κB signaling pathway through Akt1 and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2025, 51(3): 590-598.

Effect of miR-26a-5p on apoptosis of human rheumatoid arthritis fibroblast-like synovial cells through JAK2/STAT3 signaling pathway

RichHTML

PDF (PC)

Like

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 8

References 21

Related Articles 15

Metrics

Comments

Recommended 10