Page 126 - 《运动与健康科学》(英文)2024年第2期

TaggedAPTARAEnd248 C.S. Norevik et al.
power of 0.75, VO 2peak hence being expressed as mL/kg 0.75 /min. under anesthesia (5% induction and 2.5% maintenance
For more details, see the Supplementary Materials.TaggedAPTARAEnd isoflurane).TaggedAPTARAEnd

TaggedAPTARAH22.6. Rat donor plasma collectionTaggedAPTARAEnd
TaggedAPTARAH22.10. EchocardiographyTaggedAPTARAEnd
TaggedAPTARAPBased on the findings from cell culture experiments, blood
TaggedAPTARAPEchocardiography was performed using the Vevo 3100
collection for the exercise-trained rats was performed 3 h after
Imaging System (FUJIFILM VisualSonics, Toronto, ON,
the last exercise session. Donor rats were deeply anesthetized
Canada). For more details, see the Supplementary Materials.TaggedAPTARAEnd
(5% isoflurane) and their entire blood volumes were obtained
by cardiac puncture of the left ventricle. The blood was
collected in citrate tubes, which were centrifuged (2200 rela- TaggedAPTARAH22.11. Brain tissue collection and sectioningTaggedAPTARAEnd
tive centrifugal force (RCF) for 10 min at +4˚C) before plasma
TaggedAPTARAPThe rats given intraperitoneal injections of BrdU were
was pooled and stored at 80˚C until use. Approximately
transcardially perfused with Ringer’s solution (0.85% NaCl,
6 mL of plasma were collected from each rat.TaggedAPTARAEnd
0.025% KCl, 0.02% NaHCO 3 ; pH 6.9) and 4% paraformalde-
hyde (Merck kGaA, Darmstadt, Germany) using a Unified
TaggedAPTARAH22.7. Enzyme-linked immunosorbent assay in rat donor plasmaTaggedAPTARAEnd
Masterflex Drive peristaltic pump (Thermo Fisher). Brains
TaggedAPTARAPPlasma from exercised and sedentary donors was were post-fixated in 4% paraformaldehyde for 24 h at 4 ˚C and
analyzed for brain-derived neurotrophic factor (BDNF) then transferred to 2% dimethyl sulfoxide in 0.1 M phosphate
using a BDNF Rat enzyme-linked immunosorbent assay Kit buffer with 20% glycerol (Sigma-Aldrich) and stored at 4 ˚C
(Invitrogen, Carlsbad, CA, USA) on a fully automated until sectioning. The rats given intraperitoneal injections of
enzyme-linked immunosorbent assay system (Dynex DS2; saline were similarly anesthetized with 5% isoflurane in a
Dynex Technologies, Chantilly, VA, USA) according to the chamber and weighed before exsanguination and heart exci-
manufacturers’ instructions. For more details, see the sion for collecting and snap freezing the tissues.TaggedAPTARAEnd
TaggedAPTARAPThe paraformaldehyde-fixed brains were sectioned serially
Supplementary Materials.TaggedAPTARAEnd
at 40 mm in a coronal plane using a freezing microtome
(Microm HM430; Thermo Fisher Scientific). The brains were
TaggedAPTARAH22.8. Cytokine assay in rat donor plasmaTaggedAPTARAEnd
cut in 6 equally spaced series, where 5 of the series were
TaggedAPTARAPDuplicate samples of pooled plasma from exercised and
stored at 4 ˚C in tubes with 2% dimethyl sulfoxide in 0.1 mol/
sedentary donors (3 pools from 16 donors each) were analyzed
L phosphate buffer with 20% glycerol, and 1 series was
using a Bio-Plex Pro Rat Cytokine 23-Plex Assay (Bio-Rad
mounted on SuperFrost Plus Adhesion Slides (Thermo Fisher
Laboratories, Hercules, CA, USA) according to the manufac-
Scientific) for histological orientation. For more details, see
turer’s instructions. The plate was read using a Luminex 200
Supplementary Materials.TaggedAPTARAEnd
System (Luminex Corp., Austin, TX, USA) and data was
analyzed using the Bio-Plex Manager software (Version 6.1;
Bio-Rad Laboratories). For more details, see the Supplemen- TaggedAPTARAH22.12. Immunohistochemical staining and quantification for
tary Materials.TaggedAPTARAEnd amyloid plaque analysisTaggedAPTARAEnd
TaggedAPTARAPAmyloid plaques were visualized in untreated and
TaggedAPTARAH22.9. Plasma injectionsTaggedAPTARAEnd
treated later-stage AD rats using the anti-Ab mouse mono-
TaggedAPTARAPThe AD rats were treated with either saline, ExPlas, or clonal antibody, McSA1 (Supplementary Table 3) specific
SedPlas via intravenous tail vein injections over the course of for aggregates of 39- to 43-amino-acid-long Ab peptides.
6 weeks. The number of injections done in the study by Villeda Sections were incubated with the primary antibody,
et al. 17 was adapted to fit our longer treatment period. There McSA1, overnight before incubation with the secondary
were 14 injections in total, and they were administered in alter- antibody, biotinylated goat anti-mouse. The sections were
nating fashion between 2 and 3 times per week. The injection finally incubated in the avidin-biotin complex (ABC;
volume used was the maximal volume recommended for intra- Vectastain ABC kit; Vector Laboratories, Newark, CA,
28
venous injections for rats, which is 2 mL/kg body weight. TaggedAPTARAEnd USA), mounted on SuperFrost Plus Adhesion Slides, and
TaggedAPTARAPTo obtain a robust marker of cell proliferation, half of the left to dry overnight on a heating plate. The mounted
AD rats (ExPlas = 7, SedPlas = 8, saline = 7, all male) were sections were then de-fatted in xylene and coverslipped
given intraperitoneal injections of 5-bromo-2’-deoxyuridine using Entellan mounting medium (Sigma-Aldrich).TaggedAPTARAEnd
(BrdU; ab142567; Abcam, Cambridge, UK, diluted in 0.9% TaggedAPTARAPGlass slides were digitalized using Slide Scanner Axio
sterile saline and 0.02% dimethyl sulfoxide) calculated to give Scan.Z1 and processed using the software Zen 2.6 (Carl
a dose of 50 mg/kg. The other half of the AD rats (ExPlas = 7, Zeiss Microscopy GmbH, Jena, Germany). A total of 7
SedPlas = 6, saline = 9, all male) were given intraperitoneal brain sections from 1 of the 6 series (intersection distance
injections of a weight-adjusted volume of saline. The intrave- 240 mm), approximately within the bregma range from
nous injection was immediately followed by the intraperito- 3.14 to 4.52, 29 were analyzed for plaque pathology in
neal injection, and the last of the 14 treatments was the hippocampal and cortical areas utilizing Fiji software
administered 48 h before euthanasia. All injections were given (Version 1.53j; National Institutes of Health, Bethesda,