Effect and Mechanism of siRNAs Targeting IL-1β/TNF-α Combined with BMSCs Transplantation in Ameliorating Rheumatoid Arthritis in Rats
Abstract
:Simple Summary
Abstract
1. Background
2. Materials and Methods
2.1. Animals
2.2. Experimental Design
2.3. Design and Synthesize IL-1β/TNF-α siRNAs
2.4. Cell Culture and IL-1β/TNF-α siRNAs Transfection
2.5. In Vivo Tracing of BMSCs
2.6. Body Weight, Toe Swelling, Forced Immobility Time and Spleen Index Measurement
2.7. Elisa
2.8. Immunohistochemistry Analysis
2.9. Haematoxylin & Eosin and Safranin O-Fast Green Staining
2.10. RNA Isolation and RT-qPCR
2.11. Statistics Analysis
3. Results
3.1. IL-1β/TNF-α siRNAs and BMSCs on LPS-Induced RAW264.7 Inflammation
3.2. Body Weight Gain, Toe Swelling, Forced Swimming Immobility Time and Spleen Index
3.3. Serum IL-1β and TNF-α Concentrations
3.4. Histopathological Analysis
3.5. Immunohistochemical Analysis
3.6. Effects of siRNA, BMSCs and siRNA + BMSCs on mRNA Expression of Inflammatory Factors in Spleen Tissue
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Abbreviations
References
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Target Genes | Sequences (5′-3′) |
---|---|
IL-1β siRNA | S: GGAAGGCAGUGUCACUCAUTT As: AUGAGUGACACUGCCUUCCTT |
TNF-α siRNA | S: GGAUCUCAAAGACAACCAATT As: UUGGUUGUCUUUGAGAUCCTT |
Name. | Gene Reference Number | Sequence (5′-3′) |
---|---|---|
β-actin | NM_031144.2 | F: CCTCTGAACCCTAAGGCCAA R: GTCTCCGGAGTCCATCACAA |
IL-1β | NM_031512.2 | F: GGGATGATGACGACCTGCTA R: TGTCGTTGCTTGTCTCTCCT |
TNF-α | X66539.1 | F: GGTCCCAACAAGGAGGAGAA R: CTCCTCTGCTTGGTGGTTTG |
NF-κB | AF079314.2 | F: CGATCTGTTTCCCCTCATCTTTCC R: TGCGTCTTAGTGGTATCTGTGCTTCTC |
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Pan, S.; Wang, L.; Wu, B.; Xing, H. Effect and Mechanism of siRNAs Targeting IL-1β/TNF-α Combined with BMSCs Transplantation in Ameliorating Rheumatoid Arthritis in Rats. Vet. Sci. 2022, 9, 531. https://doi.org/10.3390/vetsci9100531
Pan S, Wang L, Wu B, Xing H. Effect and Mechanism of siRNAs Targeting IL-1β/TNF-α Combined with BMSCs Transplantation in Ameliorating Rheumatoid Arthritis in Rats. Veterinary Sciences. 2022; 9(10):531. https://doi.org/10.3390/vetsci9100531
Chicago/Turabian StylePan, Shifeng, Lijun Wang, Bingxing Wu, and Hua Xing. 2022. "Effect and Mechanism of siRNAs Targeting IL-1β/TNF-α Combined with BMSCs Transplantation in Ameliorating Rheumatoid Arthritis in Rats" Veterinary Sciences 9, no. 10: 531. https://doi.org/10.3390/vetsci9100531