Aktivitas Antifibrotik Ekstrak Etanol Nerium indicum Mill terstandar 5 Alfa-Oleandrin dalam Menghambat Sintesis Kolagen Fibroblas Keloid

ANNISA NURUL PRATIWI SUDARMADI, Prof. Dr. Mae Sri Hartati W., M.Si., Apt. , drg. Fara Silvia Yuliani, M.Sc

2018 | Skripsi | S1 PENDIDIKAN DOKTER

Latar Belakang: Keloid adalah lesi kulit abnormal akibat proses hiperproliferasi, aktivitas migrasi fibroblas yang berlebihan dan peningkatan sintesis kolagen. Jaringan keloid memiliki fenotip berbeda dari fibroblast kulit normal dengan karakteristik mirip sel kanker seperti mampu bereplikasi bahkan dapat menghindari sinyal antiproliferatif. Begitu banyak metode yang ditawarkan untuk terapi keloid, meskipun demikian terapi dengan efek samping minimal masih terus dikembangkan, termasuk dengan tanaman herbal. Nerium indicum (N. indicum) adalah tanaman yang diketahui memiliki banyak manfaat sebagai anti kanker, anti inflamasi, dan anti mitotik. Penelitian sebelumnya memperlihatkan kandungan utama dari Nerium indicum yaitu 5α-Oleandrin yang diketahui dapat menghambat sintesis kolagen fibroblas keloid, menghambat proliferasi fibroblas keloid serta menghambat aktivitas migrasi fibroblas. Tujuan: Penelitian ini bertujuan untuk mengkaji efek antifibrotik ekstrak etanol Nerium indicum terstandar 5α-Oleandrin dalam menghambat sintesis kolagen fibroblas keloid pada kultur sel fibroblas keloid dengan metode pewarnaan Sirius Red. Metode: Menggunakan metode eksperimental laboratorium dengan subjek kultur sel fibroblas keloid dan bahan uji ekstrak etanol terstandar Nerium indicum terstandar 5α-Oleandrin dalam menghambat sintesis timbunan kolagen fibroblas keloid. Uji sitokimia dilakukan pada kultur sel fibroblas keloid dengan pemberian enam serial konsentrasi ekstrak dengan waktu inkubasi 72 jam dengan dua kali percobaan. Viabilitas kultur sel fibroblas keloid diuji dengan pewarnaan Sirius Red dan dibaca absorbansinya dengan menggunakan ELISA Reader. Hasil : Ekstrak etanol N. indicum terstandar 5α-Oleandrin mampu menghambat sintesis timbunan kolagen kultur sel fibroblas keloid seiring dengan meningkatnya besar konsentrasi yang diberikan, persentase penghambatan sintesis kolagen juga meningkat. Analisis nilai korelasi ditemukan moderat (r=0.314 dan p=0.544) pada percobaan pertama. Nilai korelasi pada percobaan kedua juga ditemukan kuat (r=0.551 dan p=0.257). Dengan menggunakan analisis regresi probit didapatkan nilai IC50 percobaan pertama yaitu 0.055 µg/ml dan 0.053 µg/ml pada percobaan kedua dengan pada waktu inkubasi yang sama yaitu 72 jam. Kesimpulan : Ekstrak Etanol Nerium indicum terstandar 5α-Oleandrin dapat menghambat sintesis timbunan kolagen kultur fibroblas keloid dengan nilai IC50 ekstrak yaitu 0.055 µg/ml pada percobaan pertama dan 0.053 µg/ml pada percobaan kedua dengan waktu inkubasi yaitu 72 jam. Kata Kunci: Nerium indicum, sintesis kolagen, Antifibrotik, Keloid, Kolagen

Background : Keloid is abnormal skin lesion due to hyperproliferation, excessive migration activity of fibroblast and increased collagen synthesis. Keloid tissue has a different phenotype with normal fibroblasts even has cancer-like cell characteristics such as being able to replicate by its own by avoid antiproliferative signals. There are many methods are offered for keloid therapy, but therapy with minimal side effects is still being developed, including with herbs. Nerium indicum is a plant that is known to have many benefits as anti-cancer, anti-inflammatory, and anti-mitotic. Nerium indicum can be used as an antifibrotic agent because it has the activity to inhibit collagen accumulation. Previous studies have shown that the main content of Nerium indicum is 5α-Oleandrin which is known is able to inhibit the synthesis of collagen fibroblasts, the proliferation of keloid fibroblasts and fibroblasts migration activity. Objectives : This study aims to examine the antifibrotic effect of standardized ethanol extract of Nerium indicum 5-α Oleandrin in inhibiting the synthesis of collagen fibroblast in keloid fibroblast cell cultures by Sirius Red staining method. Methods : Used laboratory experimental method with keloid fibroblast cell culture subject and standardized ethanol of 5α-Oleandrin Nerium indicum in inhibiting the synthesis fibroblasts collagen. The cytochemical test was performed on cultured fibroblast cells of keloid by giving six serial concentrations extract with a 72 hour incubation time on two trials. The viability of culture of fibroblast cells of keloids was tested by Sirius Red staining and read absorbance by using ELISA Reader. Result : Standardized ethanol extract of 5α-oleandrin Nerium indicum is able to inhibit the synthesis of collagen in fibroblasts keloid culture along with the increased concentration of ethanol extract of standardized 5α-Oleandrin Nerium indicum given, the percentage inhibition also increased. The analysis of correlation values was found to be moderate (r=0.314 and p=0.544) in the first trial. The correlation value in the second experiment was also found to be strong (r=0.551 and p=0.257). By using probit regression analysis, the first experimental IC50 values were 0.055 μg / ml and 0.053 μg / ml in the second experiment with at the same incubation time of 72 hours. Conclusion : Ethanol 5α-Oleandrin standard of Nerium indicum extract can inhibit the synthesis of collagen culture collagen fibroblast collected with IC50 extract of 0.055 μg / ml in the first experiment and 0.053 μg / ml in the second experiment with the same incubation time of 72 hours. Keywords : N. indicum, collagen synthesis, antifibrotic, keloid, collagen

Kata Kunci : Nerium indicum, sintesis kolagen, Antifibrotik, Keloid, Kolagen/ N. indicum, collagen synthesis, antifibrotic, keloid, collagen