Lactobacillus Plantarum IS 10560 Supplementation in Women With Functional Constipation: Molecular Profile of Fecal Microbiome With NGS Method
Phase: PHASE2
Conditions: Constipation - Functional
Sponsor: Fakultas Kedokteran Universitas Indonesia

PROPOSAL 
29 January 2018 
 
TITLE 
Lactobacillus Plantarum IS 10560 Supplementation in Women With Functional 
Constipation: Molecular Profile of Fecal Microbiome With NGS Method 
INVESTIGATORS 
 
1. Prof. Dr. dr. Murdani Abdullah, SpPD-KGEH 
2. dr. Hasan Maulahela, SpPD 
3. dr. Amanda Pitarini Utari, SpPD 
4. Pratiwi Dyah Kusumo, S.Si., M.Biomed 
5. Prof. dr. Amin Soebandrio, Ph.D, Sp.MK 
6. Prof. Dr. dr. Dadang Makmun, SpPD-KGEH 
 
BACKGROUND 
Functional constipation as a part of Irritable Bowel Disorder is a physiological 
malfunctioning mechanism that occurs in the digestive tract where one of the main 
functions of the luminal gastrointestinal (intestinal) microbiota experiences a balance 
disorder that has an impact on the disruption as well as the expression of Short Chain 
Fatty Acid (SCFA) excreted by the intestinal microbiota. The imbalance of microbiota in 
the digestive tract will affect metabolites of Short Chain Fatty Acid (SCFA) as a result of 
fermentation of carbohydrates and proteins in the lower digestive tract. SCFA has a role 
in physiological processes (cytokine formation also activates nuclear factor κB (NF κB), 
maintains mucosal defence balance, neurotransmission regulation (SERT, TPH1 and 
serotonin), affects peristalsis and motility) Measurement of SCFA titers is one of the 
scientific bases It was established in the parent study to look at the mechanisms 
underlying the improvement of symptoms and signs of functional constipation with the 
supplementation of probiotics Lactobacillus plantarum IS 10506 indigenous to fermented 
West Sumatra curd. 
 
OBJECTIVE 
1. Test the role of Short Chain Fatty Acid (SCFA) in relation to improvement of 
symptoms and signs of Jakarta Women with Functional Constipation 
2. Test the concentration ratio of SCFA (Acetate, propionate and butyrate) as SCFA 
profiles of Jakarta Women with Functional Constipation with measurements GCMS 
and GCFID 
3. Test the relevance of changes in SCFA titers with supplementation probiotics during 
the 3-week period of supplementation of Lactobacillus plantarum IS 10506 
 
 
 

 
METHOD 
1. Design 
This study is a laboratory experimental study with two blind double randomized clinical 
trials using female subjects with functional constipation. Metabolite analysis of Short 
Chain Fatty Acid (SCFA) with GCMS is carried out after supplementation of indigenous 
Lactobacillus plantarum IS 10506 and placebo (2 variations of treatment) for three weeks. 
The analysis is carried out at the DKI Jakarta Regional Health Laboratory (DKI Jakarta 
Labkesda). 
 
2. Interventions 
Drug 
: Fermented milk containing probiotic Lactobacillus plantarum for three weeks 
Placebo  : Fermented milk containing placebo for three weeks 
 
3. Patients 
Inclusion Criteria 
- 
Women aged 18 to 60 years 
- 
Being declared healthy based on initial examination and the Structured Interview 
Questionnaire (SIQ) 
- 
Having the symptoms and signs of functional constipation refer to ROME-IV 
criteria  
- 
Able to communicate well  
- 
Able to consume 1 bottle of fermented milk each day for three weeks  
- 
Not using antibiotic no later than one week before supplementation 
 
Exclusion Criteria 
- 
Diagnosed with functional bowel disorder 
- 
Using anesthesia at least 4 weeks before treatment 
- 
Having a serious pathological disorder (carcinoma) 
- 
During healing phase of acute gastrointestinal disorders at least 4 weeks before 
treatment 
- 
Having severe heart disease 
- 
Taking chronic medications such as antidepressants or analgesics 
 
4. Procedure  
Samples will be taken from 62 (plus 10% DO predictions to 70 people) women from 
Jakarta community with a diagnosis of Functional Constipation based on ROME-IV 
established by the Medical Team of the Gastroenterology Department FK UI.  
 
Subjects will be given supplies of fermented milk containing probiotic or placebo 1-3 
days/time of arrival of researchers while monitoring compliance. Supplement is taken one 
bottle/day. During the supplementation of fermented milk, it should be kept in the 
refrigerator to a minimum of 80C. 
 

Functional constipation symptom improvement parameter (PAC Sym and PAC QoL), 
faecal samples, and blood samples will be collected three times (pre-interval-post).  
Faecal samples will be obtained using pot faeces then will be inserted in the cool box and 
ice pack stored in -200C freezer. From the faecal samples, SCFA titer will be analyzed by 
Gas chromatography–mass spectrometry (GCMS) and microbiome profile will be 
analyzed with NGS (Next Gen Sequencing) method with MiSEq Reporter software (MSR).  
 
5. Statistical Analysis 
For normally distributed data Kolmogorov-Smirnov, Non parametric test Mann-Whitney 
will be used to evaluate the differences between treatment and SCFA titer. Spearman Rho 
test will be performed to evaluate the correlation between variation of treatment and 
SCFA titer and functional constipation symptom improvement parameter. P value < 0.05 
is considered statistically significant. 
 
6. Ethical Committee Approval 
Approval from the Ethics Committee of the Faculty of Medicine, Universitas Indonesia 
must be obtained before starting the trial. 
 
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