Eichhornia crassipes Ameliorated Rheumatoid Arthritis by Modulating Inflammatory Cytokines and Metalloproteinase Enzymes in a Rat Model
Abstract
:1. Introduction
2. Materials and Methods
2.1. Plant Collection and Identification
2.2. Preparation of Methanol Extract and Fractions
2.3. Test Animals
2.4. Assessment of Anti-Arthritic Effect
2.5. Measurement of Arthritic Score and Paw Volume
2.6. Histopathological Evaluation
2.7. Analysis of mRNA Expression Values of the Pro-Inflammatory Cytokines TNF-α, IL-1β, IL-6, NF-κB, MMP-2, MMP-3, and VEGF
2.8. Determination of PGE2 Levels
2.9. Assessment of Hematological Parameters
2.10. Biochemical Parameters
2.11. GC-MS Investigation
2.12. Statistical Analysis
3. Results
3.1. E. crassipes Repressed Arthritic Development
3.2. E. crassipes Decreased Paw Edema
3.3. E. crassipes Attenuated Histopathological Scoring
3.4. E. crassipes Down-Regulated TNF-α, IL-1β, IL-6, and NF-κB
3.5. E. crassipes Attenuated MMP-2, MMP-3, and VEGF Expression Levels
3.6. E. crassipes Significantly Reduced PGE2 Levels
3.7. E. Crassipes Modulated Hematological Markers
3.8. E. crassipes Improved Markers of Liver Function Tests
3.9. No Nephrotoxic Effect of E. crassipes Was Observed
3.10. GC-MS Analysis
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Genes | Forward Primer | Reverse Primer | Annealing Temperature | Product Size | |
---|---|---|---|---|---|
IL-1β | 5′-CCTGCTAGTGTGTGATGTTC-3′ | 5′-GAGGTGCTGATGTACCAGTT-3′ | 58 °C | 390 | ENSRNOG0000004649 |
IL-6 | 5′-AGACTTCCAGCCAGTTGCCT-3′ | 5′-CTGACAGTGCATCATCGCTG-3′ | 60 °C | 233 | [24] |
TNF-α | 5′-CCTCTTCTCATTCCTGCTCGT-3″ | 5′-TGAGATCCATGCCATTGGCC-3′ | 60 °C | 266 | [25] |
NF-κB | 5′-CAAGGAAGAGGATGTGGGGTT-3 | 5′-AGCTGAGCATGAAGGTGGATG-3′ | 60 °C | 207 | [25] |
VEGF | 5′-GTTCAGAGCGGAGAAAGCATT-3 | 5′-CTTGCAACGCGAGTCTGTGT-3′ | 60 °C | 80 | [26] |
MMP-2 | 5′-CGAACAAGTATGAGAGCTGC-3′ | 5′-CGGTCATCATCGTAGTTGGT-3′ | 57 °C | 85 | ENSRNOG00000016695 |
MMP-3 | 5′-CCTTTTGATGGGCCTGGAAT-3′ | 5′-GTGACATCATCTGTCCATCG-3′ | 58 °C | 107 | ENSRNOG00000032626 |
GAPDH | 5′-GTCATCAACGGGAAACCCAT-3′ | 5′-CTTGCCGTGGGTAGAGTCAT-3 | 60 °C | 229 | [27] |
Hematological Parameters | Vehicle Control Group | Arthritic Control Group | Reference Drug Group | Methanol Extract | n-hexane Fraction | Ethyl Acetate Fraction |
---|---|---|---|---|---|---|
RBC | 8.29 ± 0.666 | 6.9 ± 0.6 *** | 7.9 ± 0.3 ## | 7.66 ± 0.275 # | 7.81 ± 0.32 # | 7.64 ± 0.46 # |
WBC | 9.60 ± 1.607 | 10.75 ± 0.736 | 9.7 ± 1.288 | 10.15 ± 0.717 | 9.47 ± 1.3 | 9.68 ± 1.422 |
PLT | 660.5 ± 37.05 | 776.7 ± 48.12 ** | 684.2 ± 62.86 # | 691.5 ± 42.93 # | 685.3 ± 25.46 # | 685.2 ± 42.44 # |
Hb | 11.73 ± 0.326 | 10.35 ± 0.796 *** | 11.80 ± 0.334 ### | 11.88 ± 0.354 ### | 11.75 ± 0.301 ### | 11.78 ± 0.23 ### |
Biochemical Parameters | Vehicle Control Group | Arthritic Control Group | Reference Drug Group | Methanol Extract | n-hexane Fraction | Ethyl Acetate Fraction |
---|---|---|---|---|---|---|
ALT | 41.33 ± 9.180 | 59.50 ± 3.391 *** | 47.83 ± 4.792 # | 48.83 ± 6.21 # | 45.67 ± 5.046 ## | 44.67 ± 4.590 ## |
AST | 278.3 ± 58.29 | 386.7 ± 44.57 ** | 291.0 ± 27.21 ## | 298.5 ± 41.23 ## | 307.2 ± 13.0 # | 289.7 ± 46.74 ## |
Urea | 19.83 ± 5.981 | 19.5 ± 2.665 | 17.17 ± 2.48 | 16.33 ± 3.83 | 15.83 ± 3.371 | 15.83 ± 5.345 |
Creatinine | 0.666 ± 0.081 | 0.65 ± 0.054 | 0.616 ± 0.04 | 0.65 ± 0.083 | 0.633 ± 0.081 | 0.6 ± 0.089 |
Sr. No. | Retention Time (Min) | Area % | Name of Detected Compound | Class of Phytochemical Compound | Molecular Formula | Molecular Weight (g/mol) | Structure |
---|---|---|---|---|---|---|---|
1 | 8.691 | 14.62 | 2-Hydroxy-1,4-naphthoquinone (Lawsone) | Naphthoquinone | C10H6O3 | 174.15 | |
2 | 17.200 | 12.39 | Octadecanoic acid | Fatty Acid | C18H36O2 | 284.48 | |
3 | 11.375 | 11.375 | Pentadecanoic acid | Fatty Acid | C15H30O2 | 242.39 | |
4 | 17.201 | 3.22 | Hexadecanoic acid | Fatty Acid | C16H32O2 | 256.4 | |
5 | 13.328 | 0.90 | Linoleic acid | Omega-6 Fatty Acid | C18H32O2 | 280.44 | |
6 | 23.4 | 0.85 | Vaccenic acid | Fatty Acid | C18H34O2 | 282.461 | |
7 | 15.46 | 0.80 | Malonic acid | Dicarboxylic acid | C3H4O4 | 104.0615 | |
8 | 31.51 | 0.78 | β-stigmasterol | Stigmastane | C29H50O | 414.71 | |
9 | 14.83 | 0.75 | Caprylic acid | Fatty Acid | C8H16O2 | 144.21 | |
10 | 16.12 | 0.70 | Nonanoic acid | Fatty Acid | C9H18O2 | 158.23 | |
11 | 18.92 | 0.65 | Myristic acid | Fatty Acid | C14H28O2 | 228.37 | |
12 | 28.5 | 0.50 | Squalene | Triterpenoid | C30H50 | 410.73 |
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Sattar, S.; Shabbir, A.; Shahzad, M.; Akhtar, T.; Ahmad, A.; Alnasser, S.M.; Riaz, B.; Karimullah, S.; Ahmad, A. Eichhornia crassipes Ameliorated Rheumatoid Arthritis by Modulating Inflammatory Cytokines and Metalloproteinase Enzymes in a Rat Model. Medicina 2023, 59, 1594. https://doi.org/10.3390/medicina59091594
Sattar S, Shabbir A, Shahzad M, Akhtar T, Ahmad A, Alnasser SM, Riaz B, Karimullah S, Ahmad A. Eichhornia crassipes Ameliorated Rheumatoid Arthritis by Modulating Inflammatory Cytokines and Metalloproteinase Enzymes in a Rat Model. Medicina. 2023; 59(9):1594. https://doi.org/10.3390/medicina59091594
Chicago/Turabian StyleSattar, Sara, Arham Shabbir, Muhammad Shahzad, Tasleem Akhtar, Arfan Ahmad, Sulaiman Mohammed Alnasser, Bushra Riaz, Shaik Karimullah, and Ashfaq Ahmad. 2023. "Eichhornia crassipes Ameliorated Rheumatoid Arthritis by Modulating Inflammatory Cytokines and Metalloproteinase Enzymes in a Rat Model" Medicina 59, no. 9: 1594. https://doi.org/10.3390/medicina59091594