The Suppression Effect of Kebar Extract on Endometriosis Lesion, MDA and TNF-α, Independent to VEGF:A Study in Endometriosis Mice Model

Abstract

The Suppression Effect of Kebar Extract on Endometriosis Lesion, MDA and TNF-α, Independent to VEGF:A Study in Endometriosis Mice Model Efek Supresi Ekstrak Kebar terhadap Lesi Endometriosis, MDA dan TNF-α, tidak Bergantung pada VEGF: Studi pada Model Tikus Endometriosis Abstract Yuli Trisetiyono 2 1 1 , Noor Pramono 1 , Syarief T. Hidayat Division of Reproductive Endocrinology and Infertility Department of Obstetrics and Gynecology Faculty of Medicine Universitas Diponegoro, Semarang Department of Embriology, Faculty of Veterinary Medicine Universitas Airlangga, Surabaya Objective: To investigate the effects of Kebar grass (Biophytum petersianum) extract administration to MDA serum levels, TNF-α and VEGF expression, and the extension of the endometriotic lesions in mice model. Methods: This study was experimentally designed. It was conducted in the Department of Obstetrics Gynecology, Faculty of Medicine Universitas Diponegoro and Faculty of Veterinary Medicine, Universitas Airlangga. Twenty-one mice were divided into three groups: the first group is control of 7 untreated endometriosis mice model, the second group consisted of 7 mice injected with leuprolide acetate 1mg/ kgBW single dose, and the last group consisted of 7 mice fed with Kebar grass extract 3mg/day for 14 days. MDA serum level was measured by spectrophotometry, TNF-α and VEGF expression by IHC using Rammele Scale Index (Immuno Reactive Score), whereas the extension of the endometriotic lesions was measured using computerize tracing. Results: Both Kebar grass extract and leuprolide acetate administration significantly decrease MDA serum levels in endometriosis mice model, compare with the control group, (0.09±0.02 nmol, 0.11±0.07 nmol, and 0.30±0.06 nmol, respectively; p=0.001). TNF-α expression of the group treated with Kebar grass extract was lower than leuprolide acetate and control group (2.43±1.521%, 4.86±0.458%, and 7.26±2.898%, respectively; p=0.002). However, there was no significant difference in VEGF expression among study groups (4.34±2.40%, 5.11±1.95%, and 7.40±3.49% respectively; p=0.116). Finally, the extension of the endometriotic lesions of the mouse models administered with Kebar grass extract and leuprolide acetate was smaller than the control group (0 mm 2 , 4.54±7.75 mm 2 , and 34.80±13.09 mm respectively; p=0.005). 2 Conclusions: Kebar grass extract has the effect of decreasing MDA serum levels and reducing TNF-α expression, resulted in smaller endometriotic lesions in mice, even though it does not affect VEGF expression. Keywords: biophytum petersianum, endometriosis, malondialdehyde, tumour necrosis factor-alpha, vascular endothelial growth factor. Abstrak Correspondence author. Yuli Trisetiyono. yulitrisetiyono@fk.undip.ac.id 1 , Widjiati 2 Tujuan: Mengetahui efek pemberian ekstrak Rumput Kebar (Biophytum petersianum) terhadap kadar MDA serum, ekspresi TNF-α dan VEGF jaringan, serta luas implan endometriosis pada tikus model. Metode: Penelitian eksperimental di Departemen Obstetri Ginekologi Fakultas Kedokteran Universitas Diponegoro dan Fakultas Kedokteran Hewan Universitas Airlangga, menggunakan 21 ekor mencit dibagi tiga kelompok, yaitu kelompok kontrol mencit model endometriosis yang hanya diberi pakan biasa, kelompok kedua diberi injeksi leuprolide asetat1mg/kgBB, dan kelompok ketiga diberi ekstrak rumput Kebar 3mg/hari selama 14 hari. Pemeriksaan MDA dari serum diukur dengan spektofotometri, ekspresi TNF-α dan VEGF dari implan endometriosis di peritoneum diukur secara IHC menggunakan Immuno Reactive Score, sedangkan luas implan endometriosis diukur dengan tracing komputer. Hasil: Ekstrak rumput Kebar dan leuprolide asetat secara signifikan menurunkan kadar MDA serum pada tikus model endometriosis, dibandingkan kelompok kontrol (0.09±0.02 nmol, 0.11±0.07 nmol, dan 0.30±0.06 nmol; p=0,001). Ekspresi TNF-α kelompok perlakuan rumput Kebar lebih rendah dibandingkan kelompok leuprolide asetat dan kontrol (2,43±1,521%, 4,86 ± 0,458%, dan 7,26 ± 2,898%; p=0.002). Akan tetapi tidak didapatkan perbedaan ekspresi VEGF yang signifikan pada semua kelompok perlakuan (4,34±2,40%, 5,11±1,95%, dan 7,40±3,49%; p=0,116). Luas lesi endometriosis pada tikus model yang diberi ekstrak rumput Kebar dan leuprolide asetat lebih kecil dibandingkan kontrol (0 mm , 4.54±7.75 mm 2 , dan 34.80±13.09 mm ; p=0,005). 2 Kesimpulan: Pemberian ekstrak rumput Kebar dapat menurunkan kadar MDA serum dan mengurangi ekspresi TNF-α, serta mengurangi luas lesi endometriosis pada tikus model, meskipun tidak mempengaruhi ekspresi VEGF jaringan. Kata kunci: Biophytum petersianum, endometriosis, malondialdehyde, tumour necrosis factor alpha, vascular endothelial

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