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

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Fig. 3

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Tab.4

References 21

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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

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

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