Effects of Sulawesi Propolis Extract on Lesion Growth, Apoptotic and Inflammatory Activity of the Rat Endometriosis Tissue

Effects of Tetragonula Aff. Biroi Propolis Extract on Lesion Growth, Apoptotic and Inflammatory Activity of the Rat Endometriosis Tissue

This study aims to evaluating inflammatory and apoptotic activity in rat's endometriosis lesion treated by Propolis compared to Dienogest

Study Overview

• Status: Completed
• Conditions: Endometriosis; Endometriosis; Peritoneum
• Intervention / Treatment: Drug: Propolis; Drug: Dienogest; Other: placebo

Detailed Description

Propolis has known of having various efficacy. The main benefits are as antimicrobial, antioxidant, anti-inflammatory, immunomodulatory and antiapoptotic.Propolis has even been shown to have an antibacterial effect in vitro, without requiring human immunological reactions.

The immunological effect of propolis is generated through immune suppression directly and decreasing cytokine production which stimulates immune process. Although proven to have a various efficacy, propolis can't be used as a therapeutic drug due to the difficulty of obtaining standard chemical composition as a requirement to be the drug's substances. Therefore, it is necessary to have biological composition in each propolis prepartions written down before comparing the efficacy.

Anti tumor activity by propolis, which has been tested in animal previously, showed that propolis can inhibit DNA synthesis in tumor cells, induce cell apoptosis and activate macrophage to produce factors which regulate the function of B, T, as well as NK cell.

Dienogest, is the next choice of progestin drug class. It has high selectivity binding in progesterone receptors, inhibits cytokine secretion in endometriosis stroma cell and induces apoptosis in endometriosis cell. Dienogest also has slight inhibitory effect against estradiol, so it can deminisce the symptoms due to hypoestrogenic such as decresing in bone mass density.

• Study Type: Interventional
• Enrollment (Actual): 120
• Phase: Early Phase 1

Contacts and Locations

Study Locations

• Indonesia
  • Capital Special Region
    • Jakarta, Capital Special Region, Indonesia
      • Animal Laboratory of Indonesia University

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Child; Adult; Older Adult
• Accepts Healthy Volunteers: No
• Genders Eligible for Study: Female

Description

Inclusion Criteria:

• female ratus rat type
• having normal estrus cycle

Exclusion Criteria:

• nothing

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Treatment
• Allocation: Randomized
• Interventional Model: Parallel Assignment
• Masking: None (Open Label)

Number of Arms

5

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: propolis 50; we do sonde everyday to the rat that given treatment of 50mg/kg propolis for 2, 4, and 6 weeks	Drug: Propolis; The propolis contains 20 mg in each ml. So, each rat we given 0,2 ml for 50mg/kg propolis treatment and 0,4 ml for 100mg/kg propolis treatment; Other Names: sulawesian propolis
Experimental: propolis 100; we do sonde everyday to the rat that given treatment of 100mg/kg propolis for 2, 4, and 6 weeks	Drug: Propolis; The propolis contains 20 mg in each ml. So, each rat we given 0,2 ml for 50mg/kg propolis treatment and 0,4 ml for 100mg/kg propolis treatment; Other Names: sulawesian propolis
Active Comparator: dienogest; we do sonde everyday to the rat that given treatment of 25mg/kg dienogest for 2, 4, and 6 weeks	Drug: Dienogest; 1mg/kg BW/day based on the study by Katayama et al. A DNG in 0,5% carboxymethylcellulose suspension was given orally with metal sound daily 0,2-0,4 mL per day; Other Names: visanne
Placebo Comparator: water; we do sonde everyday to the rat that given 0,2 ml water placebo for 2, 4, and 6 weeks	Other: placebo; we gave 0,2 ml to each rat everyday to negative control groups, and we gave no treatment to sham groups
Sham Comparator: sham group; after the 2nd laparotomy, we do nothing about sonde, just giving food and drink everyday	Other: placebo; we gave 0,2 ml to each rat everyday to negative control groups, and we gave no treatment to sham groups

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
mRNA expressions of Bax, Bcl, Cleaved Caspase 3, IL-1B, and PGE 2	The expression of Bax, Bcl-2, ad Caspase 3, IL-1B, PGE2 mRNA was carried out by isolating the RNA samples of endometriosis tissue based on the modified QIAamp RNA Blod Mini Handbook protocol. RNA spectrophotometric measurments and cDNA synthesis were carried out based on the Instruction Manual ReverTra Ace qPCR RT Master Mix with gDNA Remover protocol (Toyobo 2017: 2). In real-time PCR examination, the melting temperature (Tm) was determined to perform Quantitative Real-time PCR (Q-PCR) based on the QuantiTech SYBR Green PCR Handbook protocol (Qiagen 2011: 11) using a 3-step cycling cycle. The results were then compared with the standard curve and analyzed	through study completion, an average of 6 months
Mollecular docking	Twenty-three compounds contained in Propolis were tested by screening using rule of 5 (Lipinski's rule), screening results determine which compounds can enter the body and can work in endometriosis tissue. Furthermore, the three-dimensional shape is made of compounds which have passed the first stage. This process was created using the Marvin Sketch program. Furthermore, the compound that has been formed is stored as a ligand with extension pdb (protein data bank). Look for NFĸB receptor proteins, estrogen A, estrogen B, progesterone A and progesterone B in the protein data bank, which will be stored as protein. Ligands and proteins are tested using standard Lamarckian algorithms in the Autodock program. The results obtained are in the form of bond energy (ΔG), where smaller results indicate a higher likelihood of interaction	during the procedure
Lession size	In the second laparotomy (before the treatment are given) the investigators take a picture of the lession (as the beginning lession). Then the investigators give the treatment. After the treatment are done, the investigators do the necropsy to take a picture after the treatment was given to compare it with the beginning	during the procedure
The normality of estrus cycle after the treatment	Vaginal discharge of the rat obtained through vaginal lavage was fixed using 90% alcohol solution and stained using Giemsa. The specimen was examined microscopically to determined its estrous cycle	during the procedure

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Organs morpholgy after the treatment	In the necropsy, the investigators take ovarium, uterus, and peritonium to see if there's any change in morphology after the treatment was given	through study completion, an average of 6 months

Collaborators and Investigators

• Sponsor: Indonesia University
• Investigators: Principal Investigator: Wachyu Hadisaputra, Prof, dr, SpOG, Lecturer at Obgyn Department, Indonesia University

Study record dates

Study Major Dates

• Study Start (Actual): November 1, 2019
• Primary Completion (Actual): May 18, 2020
• Study Completion (Actual): May 18, 2020

Study Registration Dates

• First Submitted: April 21, 2020
• First Submitted That Met QC Criteria: April 30, 2020
• First Posted (Actual): May 5, 2020

Study Record Updates

• Last Update Posted (Actual): April 1, 2021
• Last Update Submitted That Met QC Criteria: March 26, 2021
• Last Verified: March 1, 2021

More Information

Terms related to this study

• Keywords: endometriosis; propolis
• Additional Relevant MeSH Terms: Endometriosis; Physiological Effects of Drugs; Anti-Infective Agents; Antineoplastic Agents; Hormones, Hormone Substitutes, and Hormone Antagonists; Antineoplastic Agents, Hormonal; Hormone Antagonists; Contraceptive Agents, Hormonal; Contraceptive Agents; Reproductive Control Agents; Contraceptives, Oral; Contraceptive Agents, Female; Contraceptives, Oral, Hormonal; Contraceptive Agents, Male; Dienogest; Propolis
• Other Study ID Numbers: 19-10-1269'

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No