Assessment of training effects on activity levels of alveolar macrophage in matured rats using chemiluminescent technique

Abstract

Chemiluminescence responses have been used for the evaluation of phagocyte function. In this study, to evaluate effects of training started after maturation on pulmonary immunity, the activity levels of rat alveolar macrophages (AMs) were assessed as reactive oxygen species (ROS) generating capacity, measured by lucigenin- and luminol-dependent chemiluminescence, using a parallel luminometer. One group of male Wistar rats started training at 11 weeks old and another group at 17 weeks old. The experimental period was 12 weeks, and about half of the rats were sacrificed after 6 weeks. The forced and voluntary exercises affect the mean levels of body weights and cell populations in the bronchoalveolar lavage fluid in younger animals; however, the voluntary exercise group in younger animals seemed to adapt after 12 weeks. By contrast, chemiluminescence responses in older rats observed after 6 weeks suggest that AMs are primed, and the maximum releasing activities of ROS are reduced. These changes in AM activity may be caused by the exercise and separation stresses and the rats may adapt to those stressors after 12 weeks. The chemiluminescent technique is thought to be useful to evaluate the changes of AM activity.

Effects of summer training camp on serum opsonic activity and plasma cytokine levels in female long‐distance runners

“…1, demonstrated that the OZ character changed to be more easily phagocytosed by the pooled human neutrophils. [5,11] Contrary to the results of PT, PH levels showed tendencies towards suppression in the middle of the camp and enhancement after the camp (Fig. 2).…”

“…These significant correlations mean that SOA levels are improving, i.e. shortening the time to release ROS and increments of ROS generation, [5,11] when cytokine levels are being suppressed.…”

“…We previously reported a CL measurement technique using a parallel luminometer with a cooled charge-coupled device (CCD) camera, and the parallel luminometer can measure 96 samples simultaneously with excellent reproducibility. [11] Some studies concerning the relations of SOA with specific immune reactions have been reported in this journal [12][13][14][15][16] No significant correlation between levels of SOA and immunoglobulin after acute exercise [13] and during a 21 day metabolic study [14] were reported, but significant correlations between depressed SOA levels and decreased immunoglobulin levels were observed during 20 days of training. [12] In this study, to evaluate the effects of the summer training camp on non-specific humoral immunity, we used the CL technique to measure ROS generation from pooled human neutrophils as an indicator of SOA.…”

“…The CL response was indicated by peak time (PT; min), which corresponds to the mean time from the recognition of foreign matters to the maximum release of ROS from neutrophils, and peak height (PH; photon/s), which corresponds to the maximum releasing activity of ROS. [5,11] Duplicate measurements were repeated twice for each OZ. PT and PH of LmCL for each OZ were represented as ratios to the mean values of six controls measured simultaneously.…”

“…A mixture of luminol (60 μL, 1 mmol/L), OZ (25 μL, 10 mg/mL) and HEPES-buffered saline (65 μL, pH 7.4) was simultaneously added into each well of the black microtiter plate using an automatic dispenser in a parallel luminometer (Alpha-Basic 47, Tokken Inc.). [11] Luminol-dependent CL (LmCL) was then automatically recorded for 60 min at 37°C with shaking. The CL response was indicated by peak time (PT; min), which corresponds to the mean time from the recognition of foreign matters to the maximum release of ROS from neutrophils, and peak height (PH; photon/s), which corresponds to the maximum releasing activity of ROS.…”