UJI AKTIVITAS NANOPARTIKEL RIBOSOME INACTIVATING PROTEIN Mirabilis Jalapa L. (RIP MJ-C) TERHADAP KEMATIAN SEL T47D

SHOFA KHOIRUN NIDA, Prof. Dr. Sismindari, SU., Apt

2017 | Skripsi | S1 FARMASI

Ribosome Inactivating Protein Mirabilis jalapa L fraksi asam (RIP MJ-C) merupakan protein yang memiliki aktivitas sitotoksik terhadap sel kanker. Namun karena mudah terdegradasi dalam tubuh, maka perlu difomulasi menjadi nanopartikel. Kombinasi antara kitosan rantai medium dan pektin metilasi rendah mampu membentuk kompleks nanopartikel yang mampu melindungi protein selama penghantaran, sehingga efek sitotoksik protein menjadi lebih baik. Fraksi asam Ribosome Inactivating Protein Mirabilis jalapa L (RIP MJ-C) diukur kadar protein dengan metode BCA dan diuji aktivitas pemotongan DNA superkoil. Pembentukan nanopartikel RIP MJ-C dengan metode gelasi ionik menggunakan konsentrasi kitosan 0.07%, pektin 0.02% dan RIP MJ-C 50 µg/ml. Nanopartikel RIP MJ-C yang terbentuk dikarakterisasi Entraptment Efficiency (EE), ukuran partikel, indeks polidispersitas, morfologi dan potensial zeta. Kemudian dilakukan uji sitotoksik terhadap sel T47D dengan waktu inkubasi 3 jam, 4 jam, 6 jam dan 24 jam. Nanopartikel RIP MJ-C yang terbentuk memiliki karakteristik Entraptment Efficiency (EE) 78.14%, ukuran partikel 306.2 nm, indeks polidispersitas 0.632, morfologi partikel sferis dan potensial zeta 26.4 mV. Uji sitotoksisitas terhadap sel T47D menunjukkan bahwa nanopartikel RIP MJ-C memiliki aktivitas sitotoksik lebih baik dibandingkan RIP MJ-C tanpa formulasi nanopartikel pada inkubasi 3 jam dan 4 jam. Kata Kunci : RIP MJ-C, nanopartikel, uji sitotoksisitas, waktu inkubasi.

The acidic fraction of Ribosome Inactivating Protein from Mirabilis jalapa L. (RIP MJ-C) is a protein that has cytotoxic activity against cancer cell lines. However, an appropriate delivery system is needed to protect RIP MJ-C from degradation in the body. Polyelectrolyte complex of medium chain chitosan and low methylated pectin can form nanoparticle which could protect proteins during its administration, therefore improving the cytotoxic activity of RIP MJ-C. In this study, concentration of RIP MJ-C was measured using BCA assay and the supercoiled DNA cleavage activity of RIP MJ-C was tested using agarose gel electrophoresis. Formulation of nanoparticle RIP MJ-C was done through ionic gelation method using 0.07% medium chain chitosan, 0.02% low methylated pectin, and 50 µg/ml RIP MJ-C. Resulted nanoparticle of RIP MJ-C was characterized by examining its entrapment efficiency, particle size, polydispersity index and zeta potential. Cytotoxic activity of nanoparticle RIP MJ-C to T47D cells was evaluated by MTT assay. Nanoparticle of RIP MJ-C had entrapment efficiency of 78.14%, particle size of 306.2 nm, polydispersity index of 0.632, spherical particle shape and zeta potential of 26.4 mV. Cytotoxic test of RIP MJ-C nanoparticle showed that RIP MJ-C nanoparticle has better cytotoxic activity than unformulated RIP MJ-C in incubation time of both 3 and 4 hours. Key words: RIP MJ-C, nanoparticle, cytotoxicity test, incubation time.

Kata Kunci : RIP MJ-C, nanoparticle, cytotoxicity test, incubation time.