Tesis

Kombinasi Madu Kelengkeng dan Etilen Glikol sebagai Krioprotektan terhadap Indeks Folikel Preantral dan Rasio Bax/Bcl-2 pada Ovarium Tikus = Combination of Longan Honey and Ethylene Glycol as a Cryoprotectant towards Preantral Follice Index and Bax/Bcl-2 Ratio of Rat Ovary.

Preservasi sel atau jaringan erat kaitannya dengan cryoinjury akibat pembentukan kristal es yang akan merusak struktur memban sel dan peningkatan kadar radikal bebas yang dapat memicu kejadian apoptosis. Vitrifikasi ovarium menggunakan krioprotektan intraseluler saja, seperti etilen glikol, diketahui masih memiliki efek toksik terhadap sel. Madu berpotensi sebagai krioprotektan ekstraseluler dengan cara menjaga kestabilan membran sel selama proses vitrifikasi dan kombinasinya dengan etilen glikol dapat mengurangi efek toksik dari etilen glikol tersebut. Penelitian ini bertujuan untuk memanfaatkan madu kelengkeng dalam menjaga integritas folikel preantral ovarium dan menekan laju apoptosis folikel selama vitrifikasi, melalui analisis densitas, indeks folikel, dan rasio Bax/Bcl-2. Pulasan histologis digunakan untuk menganalisis integritas folikel menggunakan pewarnaan Hematoksilin-Eosin, serta pewarnaan Imunohistokimia untuk melihat ekspresi dari protein proapoptosis dan antiapoptosis. Pengamatan dilakukan menggunakan Optilab 3.0, software Image Raster, dan ImageJ. Hasil penelitian menunjukkan bahwa pemberian madu dengan konsentrasi 7,5% yang dikombinasikan dengan etilen glikol, baik 7,5% maupun 15%, dapat mempertahankan densitas dan indeks folikel preantral intak, serta menunjukkan rasio Bax/Bcl-2 yang lebih rendah dibandingkan kontrol vitrifikasi pada folikel preantral. Oleh karena itu, pemberian madu mampu mempertahankan integritas folikel preantral dan menekan laju apoptosis folikel preantral sesudah vitrifikasi.
Kata Kunci : Apoptosis, cryoinjury, integritas folikel, madu kelengkeng, preservasi


Preservation of cells or tissues is related to cryoinjury due to the formation of ice crystals that will damage the cell and increase free radicals that can trigger apoptosis. Ovarian vitrification using single intracellular cryoprotectants, such as ethylene glycol, is known to still have a toxic effect on cells. Honey has the potential as an extracellular cryoprotectant by maintaining the stability of cell membranes during the vitrification process and their combinations can reduce the toxic effects of ethylene glycol. This study aims to utilize longan honey to maintain the integrity of the ovarian preantral follicles and reduce the rate of follicle apoptosis during vitrification, through density, follicle index, and Bax/Bcl-2 ratio analysis. Histological staining was used to analyze the integrity of the follicles using Hematoxylin-Eosin staining, and Immunohistochemical staining to see the expression of proapoptotic and antiapoptotic proteins. Observations were made using Optilab 3.0, Image Raster, and ImageJ software. The results showed that honey with a concentration of 7.5% combined with ethylene glycol, both 7.5%, and 15%, could maintain follicle density and intact preantral follicle index, and showed a lower Bax/Bcl-2 ratio than the vitrified control in the preantral follicle. Therefore, honey was able to maintain the integrity of the preantral follicles and reduce the rate of apoptosis of the preantral follicles after vitrification.
Keywords : Apoptosis, cryoinjury, follicle integrity, longan honey, preservation