Mechanism of human umbilical cord mesenchymal stem cell derived exosomes in promoting apoptosis of synovial fibroblasts in rheumatoid arthritis
10.3760/cma.j.cn141217-20240105-00007
- VernacularTitle:人脐带间充质干细胞外泌体促进类风湿关节炎滑膜成纤维细胞凋亡的作用机制
- Author:
Linkun BAI
1
;
Yazhen SU
;
Minghui ZHANG
;
Qianru LIU
;
Chengqiang ZHANG
;
Liyun ZHANG
;
Gailian ZHANG
Author Information
1. 山西医科大学第三医院（山西白求恩医院，山西医学科学院，同济山西医院）风湿免疫科，太原 030032
- Keywords:
Arthritis, rheumatoid;
Synoviocytes;
Mesenchymal;
Exosomes;
Signal transduction;
Apoptosis
- From:
Chinese Journal of Rheumatology
2024;28(11):819-828
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of hUCMSC-exos on the expression levels of HDAC in different isotypes of RA FLSs, and to elucidate the possible mechanism of hUCMSC-exos on the apoptosis of RA FLSs by regulating HDAC.Methods:hUCMSC and hUCMSC-Exos were isolated and identified. RT-qPCR was used to detect the changes in HDAC mRNA expression levels in FLSs after hUCMSC-Exos intervention, and the most affected HDAC types were identified. Western blot was used to detect the levels of FLS HDAC1 protein and the expression levels of NF-κB p65 and phospho-NF-κB p65 (Ser 536) in the blank control group, hUCMSC group, hUCMSC-Exos group, Trichostatin A (TSA) group and HDAC1 Inhibitor (Pyroxamide) group. To investigate the effects of hUCMSC-Exos on HDAC expression and NF-κB activity in FLSs. Flow cytometry was used to detect the effect of hUCMSC-Exos on the apoptosis of FLSs. ELISA was used to detect the effects of hUCMSC-Exos on the secretion of TNF-α, IL-6, IL-1β and IL-8 by FLSs. Flow cytometry and ELISA were used to detect the apoptosis level and pro-inflammatory cytokine secretion level of RA FLSs in the blank control group, NF-κB Inhibitor (pyrrolidine dithiocarbamate (PDTC) group, hUCMSC-Exos group and PDTC+hUCMSC-Exos co-intervention group. Whether inhibition of NF-κB affects the regulatory effect of hUCMSC-Exos on RA FLSs was further explored. All experimental data conforming to the normal distribution were compared by one-way ANOVA. LSD- t test was used for pin-group comparison, and independent sample t test was used for two-sample comparison. Results:Cultured primary hUCMSC were adherently grown spindle-shaped cells, and hUCMSC-Exos were saucer-shaped membranous vesicles, both of which met the identification criteria. hUCMSC-Exos reduced the expression level of HDCA1 mRNA [(0.932±0.091), t=2.19, P<0.001] and protein [(0.204±0.012), t=8.28, P<0.001] in RA FLSs, and the inhibitory effect was stronger than that of hUCMSC ( t=1.09, P=0.009) and HDAC1 ( t=11.29, P=0.013) Inhibitor. hUCMSC-Exos increased the apoptosis rate of RA FLSs [(48.68±0.84)%, t=12.33, P<0.001]. hUCMSC-Exos reduced the secretion levels of TNF-α [(29.6±1.0)pg/ml, t=10.78, P<0.001], IL-6 [(20.1±0.7)pg/ml, t=7.96, P<0.001], IL-1β [(9.28±0.23)pg/ml, t=6.14, P<0.001] and IL-8 [(108.0±3.8)pg/ml, t=1.21, P<0.001] in the supernatant of RA FLSs. hUCMSC-Exos reduced the expression level of p-NF-κB-p65/NF-κB-p65 in RA FLSs(0.351±0.024, t=17.67, P<0.001), and its inhibitory effect was stronger than that of hUCMSC (0.515±0.064, t=8.07, P=0.009) and HDAC1 inhibitor(0.411±0.033, t=2.44, P=0.04). After use of NF-κB inhibitors, hUCMSC-Exos weakened the promotion of apoptosis of RA FLSs [(29.0±0.5)%, t=10.63, P<0.001] and weakened the inhibitory effect of IL-8 secretion in the supernatant of RA FLSs [(125.5±3.2)pg/ml, t=2.63, P=0.002]. Conclusion:hUCMSC-Exos can mimic maternal cells to effectively inhibit the aberrant expression of HDAC1 in RA FLSs. hUCMSC-Exos may affect the apoptosis of RA FLSs and the secretion of pro-inflammatory cytokines by inhibiting the HDAC1/NF-κB pathway.
