Page 125 - 《运动与健康科学》(英文)2024年第2期

TaggedAPTARAEndExPlas promotes neurogenesis in AD rat brain 247
TaggedAPTARAPIn the experimental settings (c) and (d), a neurotoxic frag- Animal Research xx1-26, and European Convention for the
ment of Ab (Ab 25-35, A4559; Sigma-Aldrich, St. Louis, Protection of Vertebrate Animals used for Experimental and
MO, USA) was used to simulate “AD-like stress” in the cells. Other Scientific Purposes.TaggedAPTARAEnd
Ab 25-35 (Sigma-Aldrich) was dissolved in water to a concen-
tration of 50 mmol/L and incubated at 37˚C overnight to allow
TaggedAPTARAH22.3. Tests of cognitive functionTaggedAPTARAEnd
oligomerization before being diluted to 50 mmol/L in medium
TaggedAPTARAPA novel object recognition test was used to assess recogni-
and used to treat the cells for 24 h. In the final hypothesis-tes-
tion memory in the rats. The phases of the test, conducted in a
ting setting (d), which tested the effects of exercised plasma
circular arena, were habituation on Day 1, object familiariza-
collected at different time points in this “AD model”, HT22
tion on Days 24, and a test phase with a novel object on
cells were co-treated with 50 mmol/L Ab 25-35 (Sigma-
Day 5. A fear conditioning paradigm was used to assess asso-
Aldrich) and fetal bovine serum (Thermo Fisher Scientific)
ciative fear learning and memory. The 4-day protocol utilized
was replaced by pre- or post-exercise plasma at a concentra-
a combination of a tone as a conditioned stimulus and an elec-
tion of 1% for 24 h.TaggedAPTARAEnd
tric shock as an unconditioned stimulus. The ANY-maze video
TaggedAPTARAPFor each condition, the MTS assay was performed
system and software (ANY-Maze, Stoelting, Dublin, Ireland)
according to the manufacturer’s instructions in 25 biological
was used to track the rats. For more details, see the Supple-
replicates. Relative viability was measured as the quotient
obtained by dividing the mean absorbance of treated technical mentary Materials.TaggedAPTARAEnd
replicate wells (3 for each) by the mean absorbance of the
control wells. In addition to assessing cell viability, cell size TaggedAPTARAH22.4. Exercise training protocol for Wistar donor ratsTaggedAPTARAEnd
was measured in the experimental setting (d) by taking an
TaggedAPTARAPWistar rats were exercised according to a 6-week high-in-
image from approximately the center of each well prior to the
tensity interval training (HIIT) protocol on treadmills
MTS assay and having a colleague blinded to treatment groups
(Columbus Instruments, Columbus, OH, USA) with a 25˚
analyze the images using ImageJ (National Institutes of 26
incline. Prior to the first exercise session, animals were habit-
Health, Bethesda, MD, USA). In each replicate, the first 15
uated to the treadmills. The habituation consisted of 5 min on
cells starting from a random corner within each image were
the treadmill while turned off, followed by 8.0 m/min for
selected and the perimeter was drawn around the individual
10 min. The HIIT sessions were performed 5 times/week and
cells to measure cell size. For more details, see the Supplemen-
started with a 10-min warm-up at 6 m/min, immediately
tary Materials.TaggedAPTARAEnd
followed by 10 high-intensity intervals, each lasting 4 min,
with 2-min active resting periods. The active rest speed was
increased gradually from 6 m/min to 8 m/min during the 6-week
TaggedAPTARAH22.2. Animal modelTaggedAPTARAEnd
exercise training period. According to our research group’s
TaggedAPTARAPWe used the McGill-R-Thy1-APP transgenic rat model for
extensive experience training rats, the 4-min intervals required
AD (hAPP751 Swedish KM670/671NL and Indiana V717F the rats to run at a speed that corresponds to an exercise inten-
mutations, controlled by mouse Thy1.2 promoter) 25 along
sity of approximately 80%90% of their peak oxygen uptake
with non-transgenic littermates (wild type). For the rat model 27
(VO 2peak ). To maintain a similar workload throughout the
characterization data, see the Supplementary Materials
training period, the treadmill speed was progressively
(Supplementary Fig. 1 and Supplementary Table 1). Male
increased from the initial 12 m/min to the final 18 m/min. For
homozygous AD rats were assigned to transfusion treatment
more details, see the Supplementary Materials.TaggedAPTARAEnd
beginning at an early pre-plaque pathology stage (age = 2.20
§ 0.14 months, mean § SD; n = 20) and a later stage closer to
TaggedAPTARAH22.5. Cardiorespiratory fitness testingTaggedAPTARAEnd
the time point when extracellular plaques emerge (age = 5.00
§ 0.20 months; n = 23). The rats were randomly divided into 3 TaggedAPTARAPA graded maximal exercise protocol in an enclosed tread-
treatment groups: 1 receiving 0.9% NaCl (saline), 1 receiving mill with an indirect open circuit calorimeter system
plasma from sedentary young Wistar rats (SedPlas), and 1 (Oxymax; Columbus Instruments) was used to determine
receiving exercised plasma from young exercise-trained VO 2peak of the donor rats before and after the 6-week HIIT
Wistar rats (ExPlas). Male Wistar donor rats aged 12 months protocol. All rats were habituated to the enclosed treadmill
were either subjected to 6 weeks of exercise training (n = 48) before testing. This consisted of 5 min on the treadmill while
or remained sedentary (n = 48) in their standard housing condi- turned off, after which the speed of the treadmill was set to 8.0
tions, thus providing either ExPlas or SedPlas. For the study meters per min for 10 min. Both the habituation and the
overview and details on timing for procedures (Supplementary VO 2peak test were performed at a treadmill incline of 25˚. The
rat being tested was weighed and then put in the treadmill
Fig. 2 and Supplementary Table 2).TaggedAPTARAEnd
TaggedAPTARAPThe study was carried out in accordance with the Norwe- chamber, and the test was started with 10 min of warm-up at
gian regulations on animal experimentation. All experimental 8 m/min, after which the speed was increased by 1.8 m/min
procedures were approved by the competent authority for every 2 min, based on our previously published protocol. 27
animal research at the Norwegian Food Safety Authority The test was terminated when we observed a VO 2 plateauing
(FOTS ID 11740/2017) and are in accordance with the Norwe- despite increased workload. The absolute VO 2 values were
gian Animal Welfare Act xx1-28, Norwegian Regulations on scaled for size differences by dividing by body weight to the