Environmental fine particulate matter (PM 2.5) activates the RAW 264.7 macrophage cell line even at very low concentrations as revealed by 1H NMR.
Because of the association between inhalation of airborne particulate matter (PM) and human respiratory and cardiovascular disease, it is necessary to understand the tissue damage induced by these particles. One of the cell types principally involved in the body's reaction to PM are macrophages, which remove particles in the airway passages and the lungs through phagocytosis. In fact, when macrophages are exposed to a toxic agent such as PM, they undergo a series of changes (including variations in morphology, an increase in glycolysis, and consequent lactate production and the release of cytokines such as interleukin-6 and tumor necrosis factor-alpha) necessary to transform them from "resting" to "activated" macrophages. Because (1)H NMR is extremely useful in monitoring, noninvasively, macrophage metabolism and because this technique has never been utilized to examine macrophage activation after exposure to PM, it was the purpose of the present study to investigate the effects of PM exposure on the RAW 264.7 stabilized macrophage cell line using (1)H NMR spectroscopy. PM with a diameter <2.5 microm (PM 2.5) was utilized because a closer association to mortality and adverse respiratory health effects has been found with this fraction than with particles of a larger size. Measurements were conducted on whole cells at both 500 and 700 MHz as well as on perchloric acid extracts at 700 MHz. Significant variations in numerous metabolites were seen at very low concentrations of PM 2.5. Many of these changes point to activation of RAW 264.7 macrophages even at doses of PM 2.5 much lower than those commonly employed in cell studies. These results are particularly significant since the same concentrations of PM did not induce changes in morphology and release of cytokines in these cells. Therefore, (1)H NMR spectroscopy is an extremely sensitive probe in observing subtle variations in macrophages after exposure to PM 2.5.
Santini
Maria Teresa
MT
Laboratorio di Ultrastrutture, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy. santini@iss.it
Rainaldi
Gabriella
G
Ferrante
Antonella
A
Romano
Rocco
R
Clemente
Stefania
S
Motta
Andrea
A
De Berardis
Barbara
B
Balduzzi
Maria
M
Paoletti
Luigi
L
Indovina
Pietro Luigi
PL
D000397:Air Pollution / Q000009:adverse effects* / Q000032:analysis; D000818:Animals; D002453:Cell Cycle / Q000187:drug effects; D002457:Cell Extracts / Q000032:analysis; D002460:Cell Line; D004577:Electron Probe Microanalysis; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D015850:Interleukin-6 / Q000032:analysis; D008262:Macrophage Activation; D008264:Macrophages / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D010316:Particle Size; D010472:Perchlorates; D012384:Rome; D012680:Sensitivity and Specificity; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis
D002457:Cell Extracts; D015850:Interleukin-6; D010472:Perchlorates; D014409:Tumor Necrosis Factor-alpha