Sulistina, DewiRatna (2014) Pengaruh Rhodamin-B Terhadap Ekspresi BAX (Bcl-2 Antagonist X) Dan BCL-2 (B-cell lymphoma-2) Hypothalamus, FSH (Follicle Stimulating Hormone) Dan LH (Luteinizing Hormone) Pada Rattus norvegicus. Magister thesis, Universitas Brawijaya.
Abstract
Rhodamin B merupakan zat pewarna tekstil mengandung senyawa klorin (Cl-), pengalkilasi (CH3-CH3), poli aromatic hidrokarbon (PAH) mengaktivasi enzim sitokrom P-450 serta struktur quinon yang sangat redoks serta menyebabkan pembentukan Reactive Oxygen Spesies (ROS). Peningkatan ROS menginduksi apoptosis dari jalur intrinsik. Ketidakseimbangan rasio antara BAX dan BCL-2 menstimulasi apoptosis pada nucleus arkuata dan ventromedial hypothalamus (Supraoptik, Tuberal, Arkuatus, Periventrikular, Paraventrikularis). Akibat apoptosis di hypothalamus mengakibatkan penurunan sekresi GnRH sehingga mempengaruhi hipofisis anterior dalam mensekresi FSH dan LH. Dengan berkurangnya kadar FSH dan LH sampai pada target organ tidak memadai untuk mendukung perkembangan gonad. Penelitian ini bertujuan untuk membuktikan Rhodamin B meningkatkan ekspresi BAX (Bcl-2 Antagonist X) dan menurunkan ekspresi BCL-2 (B-cell lymphoma-2) di Hypothalamus, menurunkan kadar FSH (Follicle Stimulating Hormone) dan LH (Luteinizing Hormone) pada darah tikus putih Rattus norvegicus. Metode penelitian adalah “True Experimental Design” dengan desain “ post test only control group” menggunakan dua puluh delapan tikus betina Rattus norvegicus galur Wistar umur 10-12 minggu dibagi menjadi 4 kelompok yaitu kelompok kontrol, kelompok dosis 4,5 mg/200 g BB, kelompok dosis 9 mg/200 g BB dan kelompok dosis 18 mg/200 g BB. Pemberian Rhodamin B dilakukan personde selama 36 hari. Sebelum tikus di euthanasia dilakukan swab vagina untuk menentukan siklus estrus, dilakukan euthanasia pada fase proestrus. Kemudian diambil sampel jaringan hypothalamus otak tikus dan darah untuk pemeriksaan ekspresi BAX dan BCL-2 menggunakan metode immunohistokimia sedangkan untuk pemeriksaan kadar FSH dan LH menggunakan metode ELISA. Pada uji Univariant Analysis of Variance dengan tingkat kepercayaan 0,05 didapatkan hasil bahwa ada perbedaan bermakna rerata ekspresi total BAX, ekspresi BCL-2, kadar FSH dan LH keempat kelompok sampel pengamatan. Analisis hasil penelitian mengunakan uji BNT dengan tingkat kepercayaan 0.05 menunjukkan bahwa ada perbedaan yang bermakna secara statistik (p-value 0.05) ekspresi total BAX antara kelompok kontrol dengan kelompok perlakuan 4,5 mg, 9 mg, 18 mg. Ada perbedaan yang bermakna (p-value 0.05) ekspresi total BCL-2 antara kelompok kontrol dengan kelompok perlakuan 4,5 mg, 9 mg, 18 mg. Perbandingan antara kelompok kontrol dengan dosis 4,5 mg menunjukkan adanya penurunan akan tetapi tidak bermakna secara statistik. Begitu pula pada dosis 4,5 mg dengan dosis 9 mg menunjukkan adanya penurunan akan tetapi tidak bermakna secara statistik. Sedangkan perbandingan antara kelompok kontrol dengan dosis 18 mg menunjukkan adanya penurunan dan bermakna secara statistik. Ada perbedaan yang bermakna (p-value 0.05) kadar FSH antara kelompok kontrol dengan kelompok perlakuan 4,5 mg, 9 mg, 18 mg. Perbandingan antara kelompok kontrol dengan dosis 4,5 mg, 9 mg, 18 mg menunjukkan adanya penurunan dan bermakna secara statistik. Ada perbedaan yang bermakna (p-value 0.05) kadar LH antara kelompok kontrol dengan kelompok perlakuan 4,5 mg, 9 mg, 18 mg. Perbandingan antara kelompok kontrol dengan dosis 4,5 mg, 9 mg, 18 mg menunjukkan adanya penurunan dan bermakna secara statistik. Dapat disimpulkan bahwa Rhodamin B pada penelitian ini meningkatkan ekspresi BAX dan menurunkan ekspresi BCL-2 Hypothalamus, menurunkan kadar FSH dan LH pada Rattus norvegicus.
English Abstract
Rhodamine B is a textile dye compounds containing chlorine (Cl-), alkylating (CH3-CH3), Poly Aromatic Hydrocarbons (PAH) activates enzyme cytochrome P-450 as well as structure of quinone which is very redox that lead to formation of Reactive Oxygen Species (ROS). ROS increasing induce apoptosis of intrinsic pathway. imbalance ratio between BAX and BCL-2 stimulates apoptosis in arcuate nucleus and ventromedial hypothalamus (Supraoptic, Tuberal, Arcuate, Perventricular, Paraventricularis). As a result of apoptosis in hypothalamus causes GnRH secretion decreasing and thus affects anterior pituitary in secreting FSH and LH. This reducing level of FSH and LH to target organ is not adequate to support development of gonads. objective of research is to verify that Rhodamin B can increase expression of BAX (Bcl-2 Antagonist X), can reduce expression of BCL-2 (B-cell lymphoma-2) in Hypothalamus, and can decrease content of FSH (Follicle Stimulating Hormone) and LH (Luteinizing Hormone) in white blood of mouse Rattus norvegicus. Research method was “True Experimental Design”. selected design was “ post test only control group” using twenty-eight Wistar female Rattus norvegicus mouse age of 10-12 weeks. mouse were assigned into 4 groups of dose treatment including control, dose 4.5 mg/200 g body weight, dose 9 mg/200 g body weight, and dose 18 mg/200 g body weight. y were administrated Rhodamine B personde for 36 days. Before mouse had been euthanasia, vaginal swabs did first to determine estrous cycle, euthanasia performed on proestrus phase. mouse brain hypothalamus tissue samples and blood samples was taken to examined expressions of BAX and BCL-2 are analyzed with immunohistochemical method while contents of FSH and LH are analyzed with ELISA. Result of Univariate Analysis of Variance at confidence level of 0.05 had indicated that average of total expressions of BAX and BCL-2 and average of total contents of FSH and LH are significantly different between four observation sample groups. Result of analysis using BNT at confidence level of 0.05 had shown that difference of total expression of BAX between control group and treatment groups of 4.5 mg, 9 mg, and 18 mg was statistically significant (p-value 0.05). significant difference (p-value 0.05) was also observed for total expression of BCL-2 between control group and treatment groups of 4.5 mg, 9 mg, and 18 mg. comparison between control group and dose group of 4.5 mg had shown that difference of both had decreased but not statistically significant. difference between dose 4.5 mg and dose 9 mg was also reduced but also not statistically significant. comparison between control group and dose 18 mg had also indicated a reduced difference and it was statistically significant. re was a significant difference of FSH content (p-value 0.05) between control group and treatment groups of 4.5 mg, 9 mg and 18 mg. comparison between control group and dose groups of 4.5 mg, 9 mg, and 18 mg had shown a reduced difference and it was statistically significant. re was a significant difference of LH content (p-value 0.05) between control group and treatment groups of 4.5 mg, 9 mg, and 18 mg. comparison between control group and dose groups of 4.5 mg, 9 mg, and 18 mg was showing a reduced difference and it was statistically significant. It is concluded that Rhodamin B has been verified as capable to increase expression of BAX, to reduce expression of BCL-2 in hypothalamus, and to decrease contents of FSH and LH in mouse Rattus norvegicus.
| Item Type: | Thesis (Magister) |
|---|---|
| Identification Number: | TES/667.254/SUL/p/041401287 |
| Subjects: | 600 Technology (Applied sciences) > 667 Cleaning, color, coating, related technologies > 667.2 Dyes and pigments |
| Divisions: | S2/S3 > Magister Kebidanan, Fakultas Kedokteran |
| Depositing User: | Budi Wahyono Wahyono |
| Date Deposited: | 07 Apr 2014 07:53 |
| Last Modified: | 07 Apr 2014 07:53 |
| URI: | http://repository.ub.ac.id/id/eprint/160190 |
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