Effect of Melittin on Metabolomic Profile and Cytokine Production in PMA-Differentiated THP-1 Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Sample Isolation and Preparation
2.2. Cell Culture and Differentiation
2.3. Cell Viability Assay
2.4. Cytokine Production
2.5. Enzyme-Linked Immunosorbent Assay (ELISA)
2.6. Metabolite Extraction
2.7. LC-MS Conditions
2.8. Data Extraction and Statistical Analysis
3. Results
3.1. BV Fractionation and Isolation of Melittin
3.2. Cytotoxicity of Melittin against Normal and PMA-Differentiated THP-1 Cells
3.3. Effect of Melittin on the Production of Pro-Inflammatory TNF-α Cytokine
3.4. Effect of Melittin on the Production of Pro-Inflammatory IL-1β Cytokines
3.5. Effect of Melittin on the Production of Pro-Inflammatory Il-6 Cytokines
3.6. Effect of Melittin on the Production of Anti-Inflammatory Il-10 Cytokines
3.7. Effect of Melittin on the Cell Metabolome
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Mass | Rt | Putative Metabolite | LPS | Mel | Mel + LPS | |||
---|---|---|---|---|---|---|---|---|
Ratio | p Value | Ratio | p Value | Ratio | p Value | |||
Arginine and proline metabolism | ||||||||
175.096 | 15.25 | L-Citrulline * | 1.825 | 0.048 | 7.482 | 0.001 | 8.657 | 0.001 |
174.112 | 26.50 | L-Arginine * | 0.255 | <0.001 | 1.111 | ns | 0.916 | ns |
240.122 | 15.55 | Homocarnosine | 0.640 | 0.005 | 1.570 | 0.002 | 1.481 | 0.001 |
145.085 | 14.52 | 4-Guanidinobutanoate | 1.122 | 0.014 | 0.792 | <0.001 | 1.611 | <0.001 |
246.133 | 13.55 | N2-(D-1-Carboxyethyl)-l-arginine | 0.295 | <0.001 | 0.951 | ns | 1.544 | <0.001 |
113.059 | 9.30 | Creatinine | 0.288 | <0.001 | 1.045 | ns | 1.234 | ns |
129.043 | 13.62 | Oxoproline | 0.135 | <0.001 | 0.192 | <0.001 | 0.182 | <0.001 |
290.123 | 15.83 | N-(L-Arginino) succinate | 0.562 | <0.001 | 1.367 | 0.006 | 0.832 | 0.040 |
132.090 | 23.04 | L-Ornithine * | 0.387 | <0.001 | 1.051 | ns | 1.177 | 0.004 |
147.053 | 13.45 | L-Glutamate * | 0.661 | <0.001 | 0.917 | ns | 0.855 | 0.026 |
398.137 | 16.01 | S-Adenosyl-l-methionine | 0.768 | ns | 5.717 | <0.001 | 4.267 | <0.001 |
115.063 | 12.14 | L-Proline * | 0.439 | <0.001 | 0.909 | ns | 1.103 | ns |
Glycolysis/TCA cycle | ||||||||
339.996 | 17.18 | D-Fructose 1,6-bisphosphate * | 1.397 | ns | 0.544 | <0.001 | 0.671 | 0.009 |
260.030 | 15.02 | D-Fructose 6-phosphate * | 1.210 | ns | 1.620 | <0.001 | 1.590 | <0.001 |
260.020 | 15.94 | D-Glucose 6-sulfate | 2.193 | 0.000 | 1.359 | ns | 1.190 | 0.018 |
260.030 | 15.83 | D-Glucose 1-phosphate * | 1.366 | 0.004 | 2.264 | <0.001 | 1.909 | <0.001 |
809.125 | 12.25 | Acetyl-CoA | 0.713 | 0.027 | 0.946 | ns | 0.792 | 0.028 |
185.993 | 15.94 | 3-Phospho-D-glycerate | 0.753 | 0.001 | 0.610 | <0.001 | 0.501 | <0.001 |
169.998 | 15.10 | Glycerone phosphate | 0.381 | <0.001 | 0.315 | <0.001 | 0.249 | <0.001 |
192.027 | 17.28 | Citrate * | 0.318 | <0.001 | 1.089 | ns | 1.297 | 0.001 |
192.027 | 18.47 | Isocitrate * | 0.489 | <0.001 | 0.580 | <0.001 | 0.466 | <0.001 |
116.011 | 14.82 | Fumarate * | 0.751 | 0.013 | 0.891 | ns | 0.677 | 0.001 |
665.125 | 13.23 | NADH * | 0.324 | <0.001 | 0.492 | <0.001 | 0.584 | <0.001 |
663.109 | 14.17 | NAD+ * | 0.482 | <0.001 | 0.923 | ns | 0.671 | <0.001 |
OXPHOS/Pentose phosphate pathway | ||||||||
506.996 | 15.76 | ATP * | 0.751 | 0.006 | 0.987 | ns | 0.585 | <0.001 |
427.030 | 14.31 | ADP * | 0.534 | <0.001 | 0.812 | 0.012 | 0.640 | <0.001 |
258.014 | 16.69 | D-Glucono-1,5-lactone 6-phosphate | 1.214 | ns | 1.207 | ns | 1.407 | 0.026 |
290.041 | 15.23 | D-Sedoheptulose 7-phosphate | 1.831 | <0.001 | 1.181 | 0.002 | 1.489 | <0.001 |
230.019 | 14.60 | D-Ribulose 5-phosphate | 0.932 | ns | 0.579 | 0.000 | 0.713 | 0.005 |
370.007 | 17.42 | D-Sedoheptulose 1,7-bisphosphate | 1.725 | 0.002 | 0.758 | ns | 1.065 | ns |
745.091 | 16.74 | NADPH * | 1.845 | 0.049 | 1.521 | ns | 0.508 | 0.048 |
743.075 | 16.51 | NADP+ | 0.693 | 0.030 | 0.764 | 0.031 | 0.820 | 0.047 |
Purine metabolism | ||||||||
151.049 | 11.82 | Guanine * | 1.300 | ns | 0.632 | 0.010 | 1.707 | 0.001 |
283.092 | 12.93 | Guanosine | 1.310 | ns | 0.619 | 0.015 | 2.185 | 0.001 |
348.047 | 14.40 | IMP * | 2.093 | ns | 2.305 | 0.021 | 1.727 | ns |
268.081 | 10.32 | Inosine * | 4.079 | <0.001 | 1.852 | 0.001 | 6.051 | <0.001 |
168.028 | 11.38 | Urate * | 0.168 | <0.001 | 1.372 | 0.001 | 1.838 | <0.001 |
136.039 | 9.74 | Hypoxanthine * | 7.345 | <0.001 | 4.852 | <0.001 | 33.584 | <0.001 |
152.034 | 10.46 | Xanthine | 0.283 | <0.001 | 1.109 | 0.043 | 1.264 | 0.003 |
284.076 | 9.72 | Xanthosine | 0.270 | <0.001 | 1.019 | ns | 1.259 | 0.015 |
135.054 | 9.22 | Adenine | 1.246 | ns | 2.237 | 0.009 | 1.649 | ns |
347.063 | 12.62 | AMP * | 0.538 | 0.002 | 0.873 | ns | 0.339 | <0.001 |
Pyrimidine metabolism | ||||||||
403.018 | 16.15 | CDP * | 0.584 | <0.001 | 0.690 | 0.001 | 0.650 | 0.002 |
111.043 | 11.38 | Cytosine | 0.684 | ns | 2.434 | ns | 3.660 | 0.021 |
482.985 | 17.60 | CTP * | 0.659 | 0.001 | 0.719 | 0.001 | 0.463 | <0.001 |
243.086 | 11.36 | Cytidine * | 0.528 | <0.001 | 2.375 | <0.001 | 3.628 | <0.001 |
483.969 | 17.02 | UTP * | 0.617 | <0.001 | 0.685 | <0.001 | 0.437 | <0.001 |
324.036 | 15.17 | UMP * | 0.721 | 0.011 | 0.630 | <0.001 | 0.589 | <0.001 |
404.003 | 15.58 | UDP * | 0.422 | <0.001 | 0.666 | 0.001 | 0.534 | <0.001 |
580.035 | 17.95 | UDP-glucuronate | 0.604 | <0.001 | 0.560 | <0.001 | 0.495 | <0.001 |
566.055 | 16.00 | UDP-glucose | 0.637 | 0.000 | 0.555 | 0.000 | 0.457 | <0.001 |
605.077 | 17.17 | GDP-mannose | 1.812 | <0.001 | 1.996 | <0.001 | 1.532 | <0.001 |
Fatty acids and metabolites C4 | ||||||||
306.256 | 12.45 | Eicosatrienoic acid * | 1.650 | 0.004 | 1.983 | ns | 1.977 | <0.001 |
328.240 | 18.27 | Docosahexaenoic acid * | 0.346 | ns | 6.658 | 0.002 | 5.846 | 0.002 |
318.219 | 4.10 | Leukotriene A4 or isomer | 1.115 | ns | 1.469 | ns | 2.183 | 0.002 |
310.287 | 17.73 | Eicosenoic acid * | 1.328 | ns | 0.477 | ns | 0.313 | 0.009 |
356.256 | 14.78 | Prostaglandin F1alpha or isomer | 0.276 | 0.007 | 0.479 | 0.024 | 0.482 | 0.048 |
336.230 | 4.55 | Prostaglandin B1 or isomer | 1.004 | ns | 1.432 | ns | 2.125 | <0.001 |
402.225 | 4.00 | 5S-HETE di-endoperoxide or isomer | 1.709 | ns | 0.850 | ns | 0.650 | 0.007 |
282.256 | 20.06 | Oleic acid * | 1.239 | 0.036 | 9.007 | <0.001 | 8.089 | <0.001 |
256.240 | 7.84 | Hexadecanoic acid isomer | 1.397 | 0.005 | 7.235 | <0.001 | 13.602 | 0.004 |
338.319 | 24.61 | Docosenoic acid * | 1.246 | 0.005 | 2.740 | <0.001 | 2.941 | <0.001 |
268.240 | 11.26 | Heptadecenoic acid | 1.362 | 0.013 | 1.782 | <0.001 | 1.211 | ns |
366.350 | 26.59 | Tetracosenoic acid * | 1.354 | 0.038 | 2.665 | <0.001 | 2.800 | <0.001 |
304.240 | 18.36 | Eeicosatetraenoic acid * | 1.049 | ns | 46.370 | <0.001 | 42.444 | <0.001 |
332.272 | 20.30 | Docosatetraenoic acid | 0.929 | ns | 25.404 | <0.001 | 21.474 | <0.001 |
242.225 | 17.97 | Pentadecanoic acid * | 0.967 | ns | 1.787 | <0.001 | 1.758 | <0.001 |
334.287 | 21.82 | Docosatrienoic acid * | 0.776 | ns | 8.679 | 0.001 | 7.684 | 0.001 |
226.193 | 15.25 | Tetradecenoic acid | 0.890 | ns | 3.570 | 0.002 | 3.367 | 0.003 |
270.256 | 12.86 | Heptadecanoic acid | 0.874 | ns | 4.801 | 0.010 | 1.854 | 0.009 |
278.225 | 17.08 | Linolenic acid * | 0.789 | 0.029 | 3.716 | <0.001 | 2.861 | <0.001 |
280.240 | 18.38 | Linoleate * | 1.105 | ns | 9.760 | <0.001 | 8.943 | <0.001 |
Glycerophospholipids C4 | ||||||||
721.467 | 12.69 | PC 32:6 | 0.672 | ns | 0.214 | 0.016 | 0.217 | 0.023 |
832.654 | 30.65 | PG 41:0 ether | 0.665 | 0.028 | 0.623 | 0.019 | 0.402 | 0.003 |
738.448 | 12.65 | PG34:6 | 0.949 | ns | 0.357 | 0.020 | 0.308 | 0.015 |
484.280 | 13.33 | Lyso PG 16:0 | 1.589 | ns | 0.449 | 0.034 | 0.283 | 0.013 |
800.449 | 12.56 | PI 32:5 | 0.885 | ns | 0.365 | 0.009 | 0.329 | 0.005 |
584.333 | 13.28 | Lyso PI 18:0 Ether | 0.941 | ns | 0.558 | 0.020 | 0.499 | 0.004 |
858.526 | 13.47 | PI 36:4 | 1.841 | ns | 0.441 | 0.025 | 0.316 | 0.027 |
814.501 | 13.55 | PI 34:4 ether | 2.088 | ns | 0.486 | 0.025 | 0.317 | 0.019 |
785.521 | 32.15 | PS 36:3 | 0.789 | 0.008 | 0.782 | 0.005 | 0.779 | 0.007 |
517.244 | 12.56 | Lyso PS 18:4 | 1.184 | ns | 2.645 | <0.001 | 1.659 | 0.022 |
833.520 | 22.16 | PS 40:7 | 1.118 | ns | 0.439 | 0.004 | 0.470 | 0.009 |
777.457 | 13.50 | PS 36:7 | 0.970 | ns | 0.432 | 0.023 | 0.371 | 0.010 |
Miscellaneous | ||||||||
131.058 | 13.88 | N-Acetyl-beta-alanine | 2.017 | 0.004 | 4.879 | 0.001 | 5.950 | 0.001 |
217.143 | 15.88 | beta-Alanyl-L-lysine | 1.543 | 0.041 | 3.816 | <0.001 | 1.705 | 0.021 |
182.079 | 13.10 | D-Sorbitol * | 0.325 | <0.001 | 2.226 | <0.001 | 4.443 | <0.001 |
180.064 | 12.66 | D-Mannose * | 0.712 | <0.001 | 1.538 | <0.001 | 1.744 | <0.001 |
180.064 | 14.19 | D-Galactose | 1.326 | ns | 3.785 | 0.001 | 4.184 | 0.001 |
221.090 | 11.16 | N-Acetyl-D-glucosamine * | 0.464 | <0.001 | 0.953 | ns | 0.806 | 0.008 |
146.069 | 14.34 | L-Glutamine * | 0.370 | <0.001 | 1.085 | ns | 1.322 | 0.002 |
129.043 | 10.45 | 5-Oxoproline * | 2.424 | 0.018 | 0.897 | ns | 1.192 | ns |
220.085 | 8.96 | 5-Hydroxy-L-tryptophan * | 7.211 | <0.001 | 1.131 | 0.010 | 4.305 | <0.001 |
204.090 | 11.14 | L-Tryptophan * | 0.190 | <0.001 | 1.189 | ns | 1.311 | 0.046 |
181.074 | 12.34 | L-Tyrosine * | 0.309 | <0.001 | 1.050 | ns | 1.265 | 0.042 |
131.095 | 10.77 | L-Leucine * | 0.217 | 0.008 | 1.038 | ns | 1.226 | 0.048 |
117.079 | 11.94 | L-Valine * | 1.615 | 0.001 | 6.492 | 0.000 | 2.822 | 0.002 |
612.152 | 16.40 | Glutathione disulfide | 0.652 | <0.001 | 1.186 | 0.047 | 1.109 | ns |
142.074 | 13.04 | Ectoine * | 1.164 | ns | 0.553 | <0.001 | 0.196 | <0.001 |
131.069 | 13.99 | Creatine * | 0.547 | <0.001 | 0.615 | <0.001 | 0.439 | ns |
103.100 | 20.48 | Choline * | 0.444 | <0.001 | 3.571 | <0.001 | 2.827 | <0.001 |
105.043 | 15.44 | L-Serine * | 1.518 | <0.001 | 1.737 | <0.001 | 1.707 | <0.001 |
121.020 | 15.41 | L-Cysteine * | 0.581 | <0.001 | 2.182 | <0.001 | 4.862 | <0.001 |
155.070 | 15.87 | L-Histidine * | 1.088 | ns | 2.089 | <0.001 | 3.122 | <0.001 |
161.105 | 12.59 | L-Carnitine * | 0.608 | <0.001 | 0.822 | 0.008 | 0.744 | 0.002 |
238.230 | 12.66 | 2-trans-Hexadecenal C4 | 0.795 | ns | 16.865 | 0.000 | 12.016 | 0.000 |
© 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Alqarni, A.M.; Ferro, V.A.; Parkinson, J.A.; Dufton, M.J.; Watson, D.G. Effect of Melittin on Metabolomic Profile and Cytokine Production in PMA-Differentiated THP-1 Cells. Vaccines 2018, 6, 72. https://doi.org/10.3390/vaccines6040072
Alqarni AM, Ferro VA, Parkinson JA, Dufton MJ, Watson DG. Effect of Melittin on Metabolomic Profile and Cytokine Production in PMA-Differentiated THP-1 Cells. Vaccines. 2018; 6(4):72. https://doi.org/10.3390/vaccines6040072
Chicago/Turabian StyleAlqarni, Abdulmalik M., Valerie A. Ferro, John A. Parkinson, Mark J. Dufton, and David G. Watson. 2018. "Effect of Melittin on Metabolomic Profile and Cytokine Production in PMA-Differentiated THP-1 Cells" Vaccines 6, no. 4: 72. https://doi.org/10.3390/vaccines6040072