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. 2023 Jun 7:19:100204.
doi: 10.1016/j.eurox.2023.100204. eCollection 2023 Sep.

Sulawesi propolis induces higher apoptotic activity and lower inflammatory activity in a rat endometriosis model

H Situmorang  1 , A Hestiantoro  1 , S Purbadi  1 , P E Wuyung  1 , R A Werdhani  1 , A Harahap  1 , W Permadi  2 , M Sahlan  3 , W Hadisaputra  1
Affiliations

Sulawesi propolis induces higher apoptotic activity and lower inflammatory activity in a rat endometriosis model

H Situmorang et al. Eur J Obstet Gynecol Reprod Biol X. .

Abstract

Background: Endometriosis has a major impact on women's quality of life. The two primary pathologies are chronic inflammation and altered apoptotic activity. Sulawesi propolis has been shown to have known anti-inflammatory and pro-apoptotic properties in other diseases.

Objective: To investigate the effects of Sulawesi propolis in the rat endometriosis model.

Methods: An autologous endometriosis model was created in 60 female Wistar rats by laparotomy. Rats were divided into four groups (n = 15 in each group): control group (CG), dienogest group (DG), propolis 50 mg/kg body weight (BW)/day (P50) group, and propolis 100 mg/kg BW/day (P100) group. Each treatment group was divided into three different treatment durations (n = 5 in each treatment group): 2, 4 and 6 weeks. After treatment, laparotomy was performed to determine endometriotic tissue growth, apoptosis [caspase-3 and Bcl-2-associated X/Bcl-2 (Bax/Bcl)] and inflammation [prostaglandin-E2 (PGE2) and interleukin-1B (IL-1B)].

Results: A significant difference was seen in endometriotic tissue growth between the P50 group and the CG, with the greatest reduction in the P50 6-week (P50-6) group, reaching 70.66% of the initial area. Highest Bax/Bcl-2 mRNA expression was shown in the P50-4 and P100-4 groups, highest caspase-3 expression was shown in the P50-2 and P50-4 groups, and lowest IL-1B expression was shown in the P50-4 group; all differed significantly from the CG. No significant difference in PGE2S mRNA was found between the groups.

Conclusion: Sulawesi propolis extract suppressed endometriotic tissue growth in the rat model by increasing apoptotic activity. The effects were time-dependent, with 50 mg/kg BW as the optimal dose.

Keywords: Apoptotic; Chronic inflammation; Endometriosis; Propolis; Rats.

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Conflict of interest statement

The authors whose names are listed immediately below certify that they have NO affiliations with or involvement in any organization or entity with any financial interest (such as honoraria; educational grants; participation in speakers’ bureaus; membership, employment, consultancies, stock ownership, or other equity interest; and expert testimony or patent-licensing arrangements), or non-financial interest (such as personal or professional relationships, affiliations, knowledge or beliefs) in the subject matter or materials discussed in this manuscript. H. Situmorang, A. Hestiantoro, S. Purbadi, P.E. Wuyung, R.A. Werdhani, A. Harahap, W. Permadi, M. Sahlan, W. Hadisaputra.

Figures

Fig. 1
Fig. 1
Study timeline.
Fig. 2
Fig. 2
Endometriotic lesion in rat models. The yellow arrow points to a smooth-surfaced, hyperaemic cystic mass surrounded by blood vessels. Bowel adhesion can also be seen (yellow arrowhead).
Fig. 3
Fig. 3
Microscopic view of endometriotic tissue in rat model, haematoxylin and eosin staining. (A) 4 × 10 enlargement of endometriosis model. (B) A cystic mass filled with leukocytes and endometrial cells, with the cyst wall consisting of a layer of cuboidal epithelium (yellow arrow), 4 × 10 enlargement. (C, D) Cystic mass surrounded by endometrial stromal (yellow arrow) and epithelial-like structure (purple arrowhead), 40 × 10 enlargement.
Fig. 4
Fig. 4
Changes in lesion area for all treatment groups. CG, control group; DG, dienogest group; P50, propolis 50 mg/kg body weight (BW)/day; P100, propolis 100 mg/kg BW/day (Mann–Whitney test).
Fig. 5
Fig. 5
Relative changes in lesion area for each treatment duration compared with the initial lesion area. CG, control group; DG, dienogest group; P50, propolis 50 mg/kg body weight (BW)/day; P100, propolis 100 mg/kg BW/day (Mann–Whitney test). The bars and tails represent the minimum, quartile (Q) 1, Q2 (median), Q3 and maximum value of each measurement.
Fig. 6
Fig. 6
Bax/Bcl-2 mRNA expression in endometriotic tissue in various treatment groups compared with the control group (CG). DG, dienogest group; P50, propolis 50 mg/kg body weight (BW)/day; P100, propolis 100 mg/kg BW/day (Mann–Whitney test). The bars and tails represent the minimum, quartile (Q) 1, Q2 (median), Q3 and maximum value of each measurement.
Fig. 7
Fig. 7
Caspase-3 mRNA expression in endometriotic tissue in various treatment groups compared with the control group (CG). DG, dienogest group; P50, propolis 50 mg/kg body weight (BW)/day; P100, propolis 100 mg/kg BW/day (Mann–Whitney test). The bars and tails represent the minimum, quartile (Q) 1, Q2 (median), Q3 and maximum value of each measurement.
Fig. 8
Fig. 8
Prostaglandin E2 (PGE2) mRNA expression in endometriotic tissue in various treatment groups compared with the control group (CG). DG, dienogest group; P50, propolis 50 mg/kg body weight (BW)/day; P100, propolis 100 mg/kg BW/day (Mann–Whitney test). The bars and tails represent the minimum,quartile (Q) 1, Q2 (median), Q3 and maximum value of each measurement.
Fig. 9
Fig. 9
Interleukin-1B (IL-1B) mRNA expression in endometriotic tissue in various treatment groups compared with the control group (CG). DG, dienogest group; P50, propolis 50 mg/kg body weight (BW)/day; P100, propolis 100 mg/kg BW/day (Mann–Whitney test). The bars and tails represent the minimum, quartile (Q) 1, Q2 (median), Q3 and maximum value of each measurement.
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