Structure identification of potential compound as selective renal anticancer isolated from Nerium indicum Mill. Leaves

Oleandrin is one of cardenolida compound isolated from an active fraction of Nerium indicum Mill leaves. (fam. Apocynaceae), which have cytotoxic effect on several human cancer cells, and also to normal cells in vitro. Another compound which was potential as renal anticancer has also been isolated from N. indicum. However, its chemical structure has not been discovered. The aim of this study was to identify the potential compound as selective to renal cancer present in the leaves of N. indicum.The potential compound was isolated from the active fraction using Preparative TLC and structure elucidation was done by using spectroscopic methods (UV, IR, MS and NMR).Base on this spectra and by comparison with oleandrin data, it was indicated that the potential compound as Renal anticancer was as 16,17-dehydrodeacetyl-5a-oleandrin.Key word: N.indicum, oleandrin, cytotoxic, renal anticancer, spectroscopi

Deteksi mekanisme apoptosis sel kanker ginjal pada pemberian senyawa 16,17-dehidrode-asetil-5alpha-oleandrin hasil isolasi daun Nerium indicum Mill

16,17-dehidrodeasetil-Sa-oleandrin nierupakan senyawa golongan kardenolida yang diisolasi dari fraksi aktif daun Nerium indicum Mill (fam. Apocynaceae). Senyawa ini bersifat sitotoksik terhadap berbagai sel kanker dan selektif terhadap sel A498 (kanker ginjal) in-vitro, namun mekanisme apoptosisnya belum diketahui. Penelitian ini bertujuan untuk mendeteksi mekanisme apoptosis sel kanker ginjal pad a pemberian senyawa 16,17dehidrodeasetil-Sa-oleandrin menggunakan metode fluorescein isothiocyanate-annexin V dan immunositokimia. Deteksi mekanisme apoptosis terhadap sel A498, dilakukan dengan metode FITC terlabel Annexin V menggunakan Flow Cytometry (FCM). Sedangkan ekspresi protein pS3 sel A498 dilakukan dengan metode immunositokimia . Hasil penelitian menunjukkan bahwa pemberian senyawa 16,17dehidrodeasetil-Sa-oleandrin dosis 3,88x10-4 mM dengan menggunakan metode FITC-annexin V dapat menaikkan persentase jumlah kematian sel antara inkubasi 24 dan 48 jam. Senyawa 16,17-dehidrodeasetil-Sa-oleandrin dosis 1,94x10-4 mM dan 3,88x10-4 mM, dapat menaikkan ekspresi protein pS3 secara bermakna (

Cytotoxic effect of n-hexane insoluble fraction obtained from kloroform extract of kembang bulan leaves [Tithonia diversifolia (Hemsley) A. Gray] on HeLa cells and mechanism detection by Hoechst 33342

Background: Tithonia diversifolia (Hemsley) A.Gray is a plant traditionally used to cure various diseases. The previous study indicated that n-hexane insoluble fraction of chloroform extract of T. diversifolia\u27s leaves showed cytotoxic effect on Hela cells. Separation of n-hexane insoluble fraction into several fractions was conducted in order to simplify the components. Therefore, compounds responsible for cytotoxic effect and their mechanism of action can be determined. . Objectives: To determine fraction having the most cytotoxic effect and its mechanism of action obtained from n-hexane insoluble fractions of T.diversifolia leaves on Hela cells. Materials and methods: Fractionation of n-hexane insoluble fraction from chloroform extract of T. diversifolia leaves was done by using vacuum liquid chromatography. By MTT assay, the cytotoxic effect of fractions on Hela cells in vitro was determined. and doxorubicin was used as positive control. Each group was tested for 3 times In = 3) and the growth inhibition percentage of Hela cells was counted. The data was analyzed by probit regression, and mechanism of action of the active fraction is conducted by staining with hoechst 33342. Results: Fractionation gave five fractions IF1-5). The ICso levels obtained of those fractions were : F1 having ICso = 32.168 :!: 12.513F2 = 11.416 :!: 0.535F3 III = 6.223 :!: 1.303F4 = 94.308 :!: 7.843and F5 = 41.740 :!: 6.381Ig/mlwhile the ICso value of doxorubicin was 0.241 :!: 0.170 Iglml. Conclusions: Fraction 3 of the n-hexane insoluble displayed the highest cytotoxic effect on Hela cells with ICso= 6.223:!:: 1.303 Iglml. Fraction 3 at the concentration of 3.93 uglml caused chromatin condensation and fragmentation of DNA on Hela cells. Key words: T. diversifolia, cytotoxic effect. Hela cells, insoluble n-hexane fraction