Ignite Creation Date:
2024-05-06 @ 4:19 PM
Last Modification Date:
2024-10-26 @ 2:07 PM
Study NCT ID:
NCT04940884
Status:
COMPLETED
Last Update Posted:
2022-07-25
First Post:
2021-06-14
Brief Title:
Supervised Exercise Training Effects on Older Community Dwellers
Sponsor:
Chang Gung Memorial Hospital
Organization:
Chang Gung Memorial Hospital
Study Overview
Official Title:
The Effects of Exercise Intervention on Health-related Physical Fitness and Circulating microRNA in the Community Senior Residents
Status:
COMPLETED
Status Verified Date:
2022-07
Last Known Status:
None
Delayed Posting:
No
If Stopped, Why?:
Not Stopped
Has Expanded Access:
False
If Expanded Access, NCT#:
N/A
Has Expanded Access, NCT# Status:
N/A
Acronym:
None
Brief Summary:
Altered circulating microRNA miRNA after physical activity reflects exercise effects on muscle performance and cardiorespiratory fitness The present work was designed to highlight associations between exercise-induced physical fitness miRNAs in community-indwelling elderly adults Baseline clinical information was assessed for community-indwelling individuals long-term followed by our community medicine research center aged 55 years near our hospital Among them participants were randomly assigned to the supervised exercise training SET and home exercise training HET groups All included subjects were instructed to walk8000 steps per day stpd which was recorded by wrist-worm smart watches SET Participants underwent 24 sessions of moderate-intensity exercise training MICT at 70 maximum predicted heart rate for 30 min in each session HET participants underwent walking activities as the above instruction Movement analysis and body composition measurements were used to assess physical fitness at baseline and 8 as well as 24 weeks after recruitment MiRNAs miR-21 miR-126 miR-146a and miR-222 were also examined at the above time point SET participants took significantly more steps per day and had greater chance to walk8000 stpd in the community compared to the SET participants during follow-up Prominent beneficial effects on physical fitness including cardiorespiratory fitness flexibility lower extremity muscle strength and body composition were noticed during and 16 weeks after stopping SET Increased miR146a and miR-126 expressions reflecting increased anti-inflammatory response and enhanced angiogenesis respectively after 8 weeks of SET However inhibited skeletal as well as cardiac muscle catabolism respectively reflecting in the increased miR-21 and miR-222 were also identified in the present work These observations may clarify short-term SET effects on lifestyle in community inhabitants and how sensitive of miRNAs to exercise-induced physiological adaptations
Detailed Description:
Design The Institutional Review Board of a tertiary care hospital approved the study IRB No 201602058A3C502 and NCT04839796 A randomized controlled trial with assessor and subject blinded study for effects of exercise regimens on ageing persons was performed from May to November 2021 All subjects provided informed consent after understanding the experimental procedures Participants were then randomly allocated to undergo 30 min of supervised exercise training SET at moderate-intensity continuous training MICT for 24 sessions in the hospital or home exercise training HET groups using a computer-generated concealed allocation schedule Data were collected by a blinded assessor prior to randomization and after completing the exercise training
Participants Community-indwelling individuals with age55 years who lived adjacent to a community hospital were surveyed Elderly adults with mini-mental state examination MMSE score 24 and negative lumbar spine as well as lower extremity degenerative joint disorder were recruited Those who had unstable clinical presentations mentioned in our previous studies or sarcopenia diagnosed based on the recommendation of the Asian Working Group for Sarcopenia were not candidates of the study We also excluded individuals with absolute contraindications for aerobic activities suggested by the American College of Sports Medicine ACSM The baseline physical component score PCS and mental component scores MCS in the Medical Outcomes Study Short Form 36 SF-36 were used to assess the quality of life QoL and were re-evaluated 24 weeks after the recruitment Baseline demographic characteristics and clinical information of included participants were carefully recorded
Exercise Training All subjects were instructed to wear a smart watch WDI08 WisDat Inc Taichung Taiwan and to take at least 8000 steps per day stpd In addition to 8000 stpd the SET participants underwent 24 sessions of supervised MICT 70 of maximum predicted HR for 30-min on a bicycle ergometer Ergoselect 150P ergoline GmbH Bitz Germany in our hospital during an 8-week period All subjects were reminded to take 8000 stpd every week during the first 8-week after recruitment by phone call and app After completing the MICT SET participants were instructed to take 8000 stpd for another 16 weeks HET participants were instructed to take 8000 stpd over the 24-week follow-up period During the latter 16-week the instruction of taking8000 stpd was not reminded in all subjects The exercise training was terminated when the subject had symptomssigns during exercise according to the ACSM guideline20 Measurement of physical fitness The wrist-worn smart watch WDI08 Wisdat Inc Taichung Taiwan recorded the every-day step count and energy expenditure in each subject and the average number of steps and energy expenditure Kcal per day in a week represented the mean every day steps and energy expenditure of a week in community during the 24-week follow-up period
The calf circumference Calf_circ was obtained by averaging the greatest Calf_circ in bilateral legs Intelligent movement analysis eFitHealth uCare Medical Electronics Co Ltd Miaoli Taiwan using interactive voices and 3D depth image guides to assess 2-min step number 5-time sit-to-stand duration and chair sit-and-reach distance Supplementary Data 1 Each of the above test was used to estimate VO2max eV O2max lower extremity muscle strength and flexibility respectively21 Body composition including total body water TBW mineral portion protein amount Prot lean body mass LBM skeletal muscle mass SKM body fat mass BFM and basal metabolic rate BMR were measured by multiple frequency bioimpedance analysis Inbody 720 Inbody Co Ltd CA USA The above measurements were documented before 8-week after and 24-week after initial visit
RNA extraction Blood sampling of 10 ml whole blood from each subject at the recruitment 8-week and 24-week from the initial visit was placed in a tube containing 32 sodium citrate Samples were then centrifuged at 360xg for 15 min which supernatant was further centrifuged at 2400xg for 20 min at room temperature of 25 to keep plasma platelet count less than 25108ml Processed plasma of 400 microL was placed in a 2mL eppendorf Eppendorf corp Hamburg Germany to mix with 1200 microL TRIzol ThermoFisher Scientific Inc Waltham MA USA and 5 microL miR-39 5x10-15 molmicroL exogenous control of C elegans as well as 2 microg 10 microgmL in plasma yeast RNA Invitrogen Carlsbad CA USA for 15 min at room temperature Another 320 microL chloroform was added and placed at room temperature for 5 min The specimen was centrifuged at 12000 xg for 15 min at 4 and 300 microL colorless fluid layer was aspirated to mix with 900 microL iced 100 ethanol overnight at -80 The prepared specimen was placed into the Direct-zol column Direct-zol RNA Miniprep Zymo Research corp Irvine CA USA and was then centrifuged at 12000 xg for 30 sec The column was then transferred to a new collection tube and was centrifuged at 12000 xg for 30 sec after mixing with 400 microL RNA wash buffer DNase I reaction mix of 80 microL DNase I enzyme 5 microLDNA digestion buffer 75 microL was added to the tube after discarding the RNA wash buffer and was incubated at room temperature for 15 min Additional 400 microL pre-wash buffer was introduced into the tube and was centrifuged at 12000 xg for 30 sec The column was then transferred into a new 15 mL RNase-free tube and was treated with 80 microL nuclease-free water at room temperature for 2min The prepared sample was centrifuged at 12000 xg for 2 min to elute RNA solution
Quantification of plasma microRNA levels miR-21 miR-126 miR-146a and miR-222 levels at the above three different time was analyzed One-step real-time quantitative polymerase chain reaction RT-qPCR was performed using a RT-qPCR system T100TM Thermal Cycler Bio-Rad Laboratories Inc Berkeley CA USA to assess plasma microRNA levels A mixture of 100 ng total RNA extraction 10 microL TaqMan master mix ThermoFisher 58 microL nuclease-free water 02 microL universal probe library 21 10 microM 01 microL RNase inhibitor self-constructed 1 microL forward primers and 1 microL reverse primers for miR-39 miR-126 and miR-146a and 2 microL cDNA template were created on ice All reactions were incubated in a 48-well plate at 95C for 3 min followed by 40 cycles of 95C for 5 sec 60C for 10 sec and 72C for 1 sec Self-constructed forward and reverse primer sequences for primary C elegans miR-39 C-miR-39 as exogenous control Human miR-126 miR-126 and miR-146a miR-146a Supplementary Table 1 were normalized by the exogenous control and were used to determine the microRNA levels expressions during the follow-up
miRCURY LNA SYBR Green PCR kit Qiagen was used to determine human miR-21 miR-21 and miR-222 miR-222 A mixture of 4 microL 5X miRCURY RT SYBR Green Reaction Buffer 2 microL 10X miRCURY RT Wnzyme Mix and 14 microL of 100 ng total RNA extraction in nuclease-free water was incubated in a 48-well plate at 42C for 60 min and followed by 95C to generate cDNA A mixture containing 2 microL C-miR-39 or miR-21 or miR-222 primers purchased from Qiagen 10 microL miRCURY SYBR Green Master Mix 2 microL nuclease-free water and 6 microL 5X generated cDNA template was incubated at 95C for 2 min followed by 40 cycles of 95C for 10 sec and 56C for 60 sec has-miR-21 and hsa-miR-222 were normalized by the C-miR-39 and were used to determine the microRNA levels expressions during the follow-up
Determination of inflammatory activity 300 microL of serum was diluted by 12 ratios Loaded 50 microL of prepared samples per well and calibrators in duplicate onto the assay plate Multiplex Human Cytokine Panel 1 Boster Biological Technology Pleasanton CA USA Each well containing antibodies captured IL-1α IL-1β IL-6 IL-10 and TNFα A mixture that contains biotinylated analyte specific antibodies is added after washing away any unbound protein The biotinylated antibodies completed the sandwich for each specific arrayed analyte After washing away unbound biotinylated antibody streptavidin horseradish peroxidase SHRP is added Following an additional wash the amount of SHRP remaining on each location of the array is proportional to the amount of the above initially captured cytokines The amount of conjugated enzyme on each location of the array is measured with the addition of a chemiluminescent substrate
Statistical analysis Data are presented as mean 95 CI or n Differences of continuous and nominal parameters and between the two groups were estimated by student-t and chi-square tests respectively Repeated measurement ANOVA was conducted to analyze differences of continuous parameters measured at the three time points in each group Pearson correlation was performed to find the relationship between physical fitness and miRNAs A p value less than 005 was considered as statistical significance
Participants Community-indwelling individuals with age55 years who lived adjacent to a community hospital were surveyed Elderly adults with mini-mental state examination MMSE score 24 and negative lumbar spine as well as lower extremity degenerative joint disorder were recruited Those who had unstable clinical presentations mentioned in our previous studies or sarcopenia diagnosed based on the recommendation of the Asian Working Group for Sarcopenia were not candidates of the study We also excluded individuals with absolute contraindications for aerobic activities suggested by the American College of Sports Medicine ACSM The baseline physical component score PCS and mental component scores MCS in the Medical Outcomes Study Short Form 36 SF-36 were used to assess the quality of life QoL and were re-evaluated 24 weeks after the recruitment Baseline demographic characteristics and clinical information of included participants were carefully recorded
Exercise Training All subjects were instructed to wear a smart watch WDI08 WisDat Inc Taichung Taiwan and to take at least 8000 steps per day stpd In addition to 8000 stpd the SET participants underwent 24 sessions of supervised MICT 70 of maximum predicted HR for 30-min on a bicycle ergometer Ergoselect 150P ergoline GmbH Bitz Germany in our hospital during an 8-week period All subjects were reminded to take 8000 stpd every week during the first 8-week after recruitment by phone call and app After completing the MICT SET participants were instructed to take 8000 stpd for another 16 weeks HET participants were instructed to take 8000 stpd over the 24-week follow-up period During the latter 16-week the instruction of taking8000 stpd was not reminded in all subjects The exercise training was terminated when the subject had symptomssigns during exercise according to the ACSM guideline20 Measurement of physical fitness The wrist-worn smart watch WDI08 Wisdat Inc Taichung Taiwan recorded the every-day step count and energy expenditure in each subject and the average number of steps and energy expenditure Kcal per day in a week represented the mean every day steps and energy expenditure of a week in community during the 24-week follow-up period
The calf circumference Calf_circ was obtained by averaging the greatest Calf_circ in bilateral legs Intelligent movement analysis eFitHealth uCare Medical Electronics Co Ltd Miaoli Taiwan using interactive voices and 3D depth image guides to assess 2-min step number 5-time sit-to-stand duration and chair sit-and-reach distance Supplementary Data 1 Each of the above test was used to estimate VO2max eV O2max lower extremity muscle strength and flexibility respectively21 Body composition including total body water TBW mineral portion protein amount Prot lean body mass LBM skeletal muscle mass SKM body fat mass BFM and basal metabolic rate BMR were measured by multiple frequency bioimpedance analysis Inbody 720 Inbody Co Ltd CA USA The above measurements were documented before 8-week after and 24-week after initial visit
RNA extraction Blood sampling of 10 ml whole blood from each subject at the recruitment 8-week and 24-week from the initial visit was placed in a tube containing 32 sodium citrate Samples were then centrifuged at 360xg for 15 min which supernatant was further centrifuged at 2400xg for 20 min at room temperature of 25 to keep plasma platelet count less than 25108ml Processed plasma of 400 microL was placed in a 2mL eppendorf Eppendorf corp Hamburg Germany to mix with 1200 microL TRIzol ThermoFisher Scientific Inc Waltham MA USA and 5 microL miR-39 5x10-15 molmicroL exogenous control of C elegans as well as 2 microg 10 microgmL in plasma yeast RNA Invitrogen Carlsbad CA USA for 15 min at room temperature Another 320 microL chloroform was added and placed at room temperature for 5 min The specimen was centrifuged at 12000 xg for 15 min at 4 and 300 microL colorless fluid layer was aspirated to mix with 900 microL iced 100 ethanol overnight at -80 The prepared specimen was placed into the Direct-zol column Direct-zol RNA Miniprep Zymo Research corp Irvine CA USA and was then centrifuged at 12000 xg for 30 sec The column was then transferred to a new collection tube and was centrifuged at 12000 xg for 30 sec after mixing with 400 microL RNA wash buffer DNase I reaction mix of 80 microL DNase I enzyme 5 microLDNA digestion buffer 75 microL was added to the tube after discarding the RNA wash buffer and was incubated at room temperature for 15 min Additional 400 microL pre-wash buffer was introduced into the tube and was centrifuged at 12000 xg for 30 sec The column was then transferred into a new 15 mL RNase-free tube and was treated with 80 microL nuclease-free water at room temperature for 2min The prepared sample was centrifuged at 12000 xg for 2 min to elute RNA solution
Quantification of plasma microRNA levels miR-21 miR-126 miR-146a and miR-222 levels at the above three different time was analyzed One-step real-time quantitative polymerase chain reaction RT-qPCR was performed using a RT-qPCR system T100TM Thermal Cycler Bio-Rad Laboratories Inc Berkeley CA USA to assess plasma microRNA levels A mixture of 100 ng total RNA extraction 10 microL TaqMan master mix ThermoFisher 58 microL nuclease-free water 02 microL universal probe library 21 10 microM 01 microL RNase inhibitor self-constructed 1 microL forward primers and 1 microL reverse primers for miR-39 miR-126 and miR-146a and 2 microL cDNA template were created on ice All reactions were incubated in a 48-well plate at 95C for 3 min followed by 40 cycles of 95C for 5 sec 60C for 10 sec and 72C for 1 sec Self-constructed forward and reverse primer sequences for primary C elegans miR-39 C-miR-39 as exogenous control Human miR-126 miR-126 and miR-146a miR-146a Supplementary Table 1 were normalized by the exogenous control and were used to determine the microRNA levels expressions during the follow-up
miRCURY LNA SYBR Green PCR kit Qiagen was used to determine human miR-21 miR-21 and miR-222 miR-222 A mixture of 4 microL 5X miRCURY RT SYBR Green Reaction Buffer 2 microL 10X miRCURY RT Wnzyme Mix and 14 microL of 100 ng total RNA extraction in nuclease-free water was incubated in a 48-well plate at 42C for 60 min and followed by 95C to generate cDNA A mixture containing 2 microL C-miR-39 or miR-21 or miR-222 primers purchased from Qiagen 10 microL miRCURY SYBR Green Master Mix 2 microL nuclease-free water and 6 microL 5X generated cDNA template was incubated at 95C for 2 min followed by 40 cycles of 95C for 10 sec and 56C for 60 sec has-miR-21 and hsa-miR-222 were normalized by the C-miR-39 and were used to determine the microRNA levels expressions during the follow-up
Determination of inflammatory activity 300 microL of serum was diluted by 12 ratios Loaded 50 microL of prepared samples per well and calibrators in duplicate onto the assay plate Multiplex Human Cytokine Panel 1 Boster Biological Technology Pleasanton CA USA Each well containing antibodies captured IL-1α IL-1β IL-6 IL-10 and TNFα A mixture that contains biotinylated analyte specific antibodies is added after washing away any unbound protein The biotinylated antibodies completed the sandwich for each specific arrayed analyte After washing away unbound biotinylated antibody streptavidin horseradish peroxidase SHRP is added Following an additional wash the amount of SHRP remaining on each location of the array is proportional to the amount of the above initially captured cytokines The amount of conjugated enzyme on each location of the array is measured with the addition of a chemiluminescent substrate
Statistical analysis Data are presented as mean 95 CI or n Differences of continuous and nominal parameters and between the two groups were estimated by student-t and chi-square tests respectively Repeated measurement ANOVA was conducted to analyze differences of continuous parameters measured at the three time points in each group Pearson correlation was performed to find the relationship between physical fitness and miRNAs A p value less than 005 was considered as statistical significance
Study Oversight
Has Oversight DMC:
None
Is a FDA Regulated Drug?:
False
Is a FDA Regulated Device?:
False
Is an Unapproved Device?:
None
Is a PPSD?:
None
Is a US Export?:
None
Is an FDA AA801 Violation?:
None