Effect of Endurance Exercise on nrf2 mRNA Expression Gene and Physical Fitness (VO2max) of Indonesian Hajj Health Officers
Phase: NA
Conditions: Physical Fitness
Sponsor: Hasanuddin University

15 May 2019 
 
 
Effect of endurance exercise on nrf2 mRNA expression gene and physical 
fitness (VO2max) of Indonesian Hajj Health Officers 
 
 
Author's name: Ismail Ahmad1*, Budu Budu2, Mochammad Hatta2, Andi Ihsan3, Dany 
Haryanto1, Riza Anshori1 
 
Affiliation: 1Postgraduate School, Doctoral Study Program, Medical Sciences, Hasanuddin University Makassar, 90245, Indonesia 
2Department of Biomolecular Medicine, Hasanuddin University Makassar, 90245, Indonesia 
3Department of Sports Health, Faculty of Sport Science, Makassar State University, 90245, Indonesia 
 
 
Contact Person: Ismail Ahmad 
Doctoral study program, Medical Sciences, Hasanuddin University  
90245 Jl. Perintis Kemerdekaan Km. 10 Makassar, Indonesia  
Tel.: +62 813 5491 6037 
Email: ismailskpns@gmail.com 
 

 
1. Background 
Indonesian Hajj Health Officers are health workers assigned by the Minister of Health of 
the Republic of Indonesia to provide services, guidance, and health protection to pilgrims during 
the Hajj pilgrimage, not it is only sufficient to have work competence in terms of knowledge, skills 
and work attitudes that are in accordance with established standards, but it is highly demanded to 
have excellent physical endurance.  An Indonesian Hajj Health Officer is required to have 3 (three) 
abilities in carrying out his duties and responsibilities in the health sector, namely (a) managerial 
ability, (b) skill ability and (c) excellent physical ability.  Excellent physical ability is one of the 
main keys to success in providing health services to Indonesian Hajj pilgrims, this is possible 
because the number of Indonesian Hajj pilgrims is very large and the length of time serving 
Indonesian Hajj Health Officers for approximately 39 days for TKHI and 76 days for PPIH.  The 
physical fitness of a Hajj Health Officer is required in addition to being in general fitness, they 
must also be demanded for specific physical fitness. Standarization of individual physical fitness 
in general can be measured by the VO2max level is one of the markers of physical fitness and an 
increase in VO2max. Regular exercise can increase of VO2max levels.  So the higher one's VO2max, 
also has good endurance. Endurance exercise consists of two groups, namely endurance cardio 
respiration (general) and muscle endurance (local). For measurement of cardiorespiratory 
endurance by VO2max.  Some of the results of research in the field biomolecular of genetic markers 
for endurance, one of which is the nrf2 gene, also known as GA-binding protein transcription 
factor β sub unit (GABPBI), which is a transcription factor in mitochondrial biogenesis.  
Mitochondrial Biogenesis and nucleus expressed by one of the nrf2 mRNA genes encoded by the 
GABPB1 gene, located on chromosome 15q212.  
 

 
2. Objectives 
The aim of the research was to examine the effects of Endurance Exercise on the Expression of 
the mRNA Nrf2 gene and Level of VO2max of Indonesian Hajj Health Officers. 
 
3. Methods 
This study was a single group quasy experiment that took place in two locations the Makassar 
Health Training Center and Makassar Haji Sudiang Dormitory. 
 
3.1 Participant 
Participants are Prospective Indonesian Hajj Health Officers as many as 30 participants consisting 
of 17 men and 13 women. Criteria for inclusion of participants include: (a) Participants from the 
Nurse profession aged 30 - 39 years (b) Pass the selection of candidates for Indonesian Hajj Health 
Officers (c) Understand the physical training program instructions in the rockport method (d) 
willing to participate in activities and all research provisions . Participants who (a) had a history 
of coronary heart disease,asthma; (b) not approved by the doctor to participate in the study due to 
a medical condition, excluded from the study. 
 
3.2 Desain and measure 
3.2.1 Exercise Protocol 
Subject (n = 30), measured VO2max level 3 times using the Multistage Fitness technique Test (MFT) 
Bleep Test method (level and feedback) before and after the 1600 meter running exercise duration 
20-30 minutes 3 times a week with a frequency of exercise 16 times, then measuring VO2max, 1 
week after exercise without running intervention. 

 
3.2.2 VO2max Measurement 
Level value and feedback from the Bleep test, then the value is converted into the VO2max based 
on the level achievement and feedback obtained, then the VO2max value  is grouped into the form 
of levels including special, excellent, good, moderate , less and very less. 
3.2.3 Measurement of mRNA NRF2 Gene Expression 
RT-PCR analyzes human RNA in the nrf2 RNA sample as the target gene and GAPDH is selected 
as the housekeeping gene. Samples were preserved in nuclease free water (Norgen biotech, Inc.), 
and RNA was extracted using the RNA isolation kit "RNAprep pure kit for blood" (Tiangen 
Biotech, Beijing). cDNA is cleaned using the "iScript cDNA sytesis kit" (Bio-Rad Laboratories, 
Inc.) cDNA kit and store at -20oC until analyzed by RT-PCR. RT-PCR is run on CFX96 touch 
(Bio-Rad Laboratories, Inc.) using the QPCR master mix "Ssofastevagreen supermix" (Bio-Rad 
Laboratories, Inc.). Specific gene primer pairs are described as follows:  
Forward: GAGAGCCCAGTCTTCATTGC; 
Reverse NRF2 Primary: TTGGCTTCTGGACTTGGAAC. 
 
3.3 Statistical Analysis Method 
Statistical analysis using SPSS Software (version 21. Inc. Chicago, IL USA) Wilcoxon sign rank 
test was used to prove the effects of endurance exercise on nrf2 mRNA expression gene and 
VO2max. Liner regression test and Spearman Rank to analyze the correlation of the nrf2 gene 
mRNA with VO2max and the Mann Whitney test to prove changes in retention VO2max.