The potentiation effect of Bawang Dayak (Sisyrinchium palmifolium L.) extract on T47D cell growth inhibition after 5-fluorouracil treatment

5-Fluorouracil (5-FU) is chemotherapy agents that used in breast cancer treatment that has toxic effects such as myelotoxicity. its effectiveness relatively low. Bawang Dayak has been widely empirically used as a breast cancer treatment. This study's aims were to determine the sensitivity of 5-FU against T47D cells, as single and its combination with EBD, to determine apoptotic induction, and the inhibitory effect of T47D cell cycle. The cytotoxic assay was using the MTT method with the concetration of 5-FU (0-600μg / mL) and EBD (0-250μg / mL). Apoptotic induction analysis were determined by PI/annexin V. Both of apoptotic and cell cycle analysis was performed by flowcytometry. The results of the 5-FU and EBD cytotoxic test on T47D cells obtained IC50 274μg/mL and 75μg/mL, respectively. The combination of 5-FU and EBD has a mild to strong synergistic effect (CI 0.829 and 0.779). its ombined treatment induced apoptosis in T47D cells by 21.33%. Combination of these combinations caused cell accumulation in G0-G1 phase of 57.11%, greater than control (41.20%). Provision of 5-FU and EBD combination treatments causes disturbance and damage to cells that cause cell cycle arrest in the G0-G1 phase and can not proceed to the next phase. The combination of 5-fluorouracil and EBD has a synergistic effect as breast cancer co-chemotherapy is able to induce apoptosis against T47D cells by triggering cell accumulation in G0-G1 phase. The extract of Bawang Dayak has a high potential to be developed as co-chemotherapy in breast cancer

UJI AKTIFITAS SERUM EKSTRAK TERPURIFIKASI DAUN APUH-APUHAN (AZOLLA MICROPHYLLA) SEBAGAI ANTIAGING

Penuaan merupakan proses menghilangnya kemampuan jaringan secara perlahan-lahan untuk memperbaiki dan mempertahankan struktur serta fungsi normalnya. Penyebab penuaan kulit dini yang paling banyak terjadi dikarenakan oleh paparan sinar matahari yang terinduksi. Penelitian ini dilakukan menggunakan ekstrak terpurifikasi daun apuh-apuhan sebagai zat aktif dalam sediaan serum dan pengujian sebagai anti-kolagenase, anti-elastase dan anti-tirosinase sebagai antiaging. Penentuan kadar flavonoid dilakukan menggunakan spektrofotometri UV-Vis dan hasil yang didapatkan kadar total flavonoid sebesar 8,222±0,03 mgQE/g. Kombinasi bahan yang digunakan dalam pembuatan serum yaitu: karbopol, trietanolamin, propilen glikol, metil paraben, propil paraben dan aquades. Hasil dari formula sediaan serum ekstrak terpurifikasi daun apuh-apuhan memiliki warna hijau jernih,tekstue semi kental, memiliki pH 5,5 dan viskositas 1310 cP hasil yang diperoleh cukup baik. Pada pengujian terhadap enzim kolaganase pada konsentrasi 500 ppm, 1000 ppm dan 2000 ppm memiliki persen penghambatan 38,91%, 45,15% dan 53,61%. Konsentrasi yang digunakan pada aktivitas enzim tirosinase, mendapatkan persentase penghambatan 42,45%, 45,68% dan 45,80%. Konsentrasi tersebut digunakan pada pengujian terhadap enzim elastase, mendapatkan hasil 62,83%, 84, 71% dan 89,24%. Sehingga dapat disimpulkan bahwa ekstrak terpurifikasi daun apuh-apuhan mempunyai aktifitas sebagai antiaging.Aging is the gradual loss of tissue's ability to repair and maintain its normal structure and function. The most common cause of premature skin aging is due to induced sun exposure. This research was conducted using purified extract of apuh-apuhan leaf as an active substance in serum preparations and testing as anti-collagenase, anti-elastase, and anti-tyrosinase as antiaging. Determination of flavonoid levels was carried out using UV-Vis spectrophotometry and the results obtained were total flavonoid levels of 8.222±0.03 mgQE/g. The combination of ingredients used in the manufacture of serum are carbopol, triethanolamine, propylene glycol, methylparaben, propylparaben, and aquades. The results of the serum preparation formula for the purified extract of apuh-apuhan leaf have a clear green color, semi-viscous texture, have a pH of 5.5 and a viscosity of 1310 cP, the results obtained are quite good. In testing the collagenase enzyme at concentrations of 500 ppm, 1000 ppm and 2000 ppm had inhibition percentages of 38.91%, 45.15% and 53.61%. The concentration used on the activity of the tyrosinase enzyme obtained the percentage of inhibition of 42.45%, 45.68% and 45.80%. These concentrations were used in testing the elastase enzyme, getting results of 62.83%, 84, 71% and 89.24%. So it can be concluded that purified extract of apuh-apuhan leaves has antiaging activity

ISOLASI ETIL P-METOKSISINAMAT DARI RIMPANG KENCUR (Kaempferia galangal L.) DAN IDENTIFIKASINYA DENGAN KROMATOGRAFI GAS SPEKTROSKOI MASSA

ABASTRAK Etil p-metoksisinamat (EPMS) adalah salah satu senyawa hasil isolasi rimpang kencur (Kaempferia galangal L.) yang merupakan bahan dasar senyawatabir surya yaitu pelindung kulit dari sengatan sinar matahari. Metode yang digunakan dalam penelitian ini adalah perkolasi. Setelah didapatkan perkolat selanjutnya dihilangkan pelarutnya hingga terbentuk Kristal dan selanjutnya dilakukan proses rekristalisasi. Hasil penelitian menunjukkan bahwa kemungkinan besar EPMS berada dalam ekstrak kencur yang diekstraksi menggunakan pelarut N-heksan dan mempunyai kelarutan dan kemurnian yang cukup tinggi yaitu sebesar 96,92%. Hal ini bisa dilihat dari kromatogram pada Kromatografi Gas Spektroskopi Massa yang menunjukkan 1 puncak yang sangat dominan. Kata kunci : EPMS, kencur (Kaempferia galanga L.), isolasi, KGSM. ABSTRACT EPMS – Ethyl p-methoxycinnamate was one of compound isolated from greater galingale rhizome (Kaempferia galanga L.) can be used as base substance of sun screen product. The method used in this research was percolation. The obtained percolate was dried until crystal was formed. The obtained crystal was later recrystallization. The result showed that greater galingale N-hexane extract possibly contained EPMS. The isolated substance had high solubility and purity (96,92%), it was seen from Gas Cromatograph Mass Spectrometer spectra that show one dominant peak. Keywords: EPMS, greater galingale (Kaempferia galanga L.), isolation, GCMS

THE ETHYL ACETATE FRACTION OF Gynura procumbens SENSITIZES WiDr COLON CANCER CELL LINE AGAINST 5-FLUOROURACIL BUT SHOWS ANTAGONISM WITH CISPLATIN

Our recent study has evaluated fraction of ethyl acetate of Gynura procumbens (FEG) as co-chemotherapeutic agent in combination with 5-fluorouracil (5-FU) and cisplatin (CISP) against colon cancer cell line, WiDr cells. This study aims to assess whether FEG show synergism with 5-FU and CISP and to evaluate its regulation on proliferation, cell cycle, and apoptosis on WiDr colon cancer cells. (3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide) MTT assay was performed to determine the growth inhibitory effect of both single (FEG, 5-FU, or CISP) and combination treatments. FEG (25-500 μg/mL),  5-FU (25-1000 μM)  and CISP (5-100 μM) inhibited cells growth in a dose dependent manner and exhibited IC50 value of 125  μg/mL, 848 mM and 43 mM, respectively. FEG sensitizes WiDr cells that was treated by 5-FU, boosting its therapeutic potential. Conversely when FEG was combined with CISP, it caused antagonism.  The antiproliferative effect of single and combination treatment was determined by studying the cell proliferation kinetics under MTT assay. Flowcytometry and (4’,6-diamidino-2-phenylindole) DAPI staining was used to disclose the mechanism of cell cycle arrest and apoptosis. FEG inhibited cell proliferation, induced G1 and S phase arrest and apoptosis. The inhibitory effect was enhanced when FEG was combined with 5-FU, differing from CISP. According to the datas obtained, FEG appeared to possess sensitizing properties, and caused cell cycle arrest and apoptosis on WiDr cells. FEG demonstrated a possibility of additive to synergism properties when combined with 5-FU but not with CISP.   Keywords: Gynura procumbens; WiDr; G1 and S phase arrest; apoptosis

EFFECT OF LATEX FROM MUSA PARADISIACA VAR. SAPIENTUM AND CARICA PAPAYA L. ON PROLIFERATION AND MIGRATION FIBROBLAST CELL NIH3T3

Objective: The objective of this research was to screen phytochemical constituents and determine the activity of latex from Musa paradisiacavar. sapientum and Carica papaya L. to the process of wound healing in NIH3T3 fibroblasts cells through observations of the proliferation andmigration of cells.Materials and Methods: Screening phytochemical compounds of latex from M. paradisiaca var. sapientum and C. papaya L. used chemical reagent.Cytotoxic activity using 3-(4,5-dimethylthiazol-2-yl)-2.5-diphenyl tetrazolium bromide (MTT) method. The proliferation test used the doubling timemethod at a susceptible incubation time of 0, 24, 48, and 72 h with a concentration of 25 μg/mL–250 μg/mL. The migration test was carried out usingthe scratch wound healing method with a concentration of 25 μg/mL–250 μg/mL in the time range of 0, 12, 24, and 48 h.Results: Phytochemical compounds contained in the latex from M. paradisiaca var. sapientum (saponin and tannin) and C. papaya L. (saponin andalkaloid). The cytotoxic assay results showed that no toxic effect for NIH3T3 fibroblasts cells (IC50 >1000 μg/mL). Cell proliferation and migrationtest results showed an increase in NIH3T3 fibroblast cell proliferation and migration process compared to controls. The concentration of 250 μg/mLof latex from M. paradisiaca var. sapientum and C. papaya L. is the best to increase of proliferation and migration process of NIH3T3 fibroblast cells.Conclusion: This study concludes that the latex of M. paradisiaca var. sapientum and C. papaya L. has the potential to increase proliferation andmigration activity of NIH3T3 cells

Gynura procumbens Prevents Chemoresistance through Inhibition MDR1 Expression on MCF-7 Breast Cancer Cell Line and Sensitizes the Cells to Doxorubicin

The long-term exposure of doxorubicin (Dox) causes enhancement in MDR1 expression that leads to breast cancer cell resistance. This protein become a serious problem in cancer treatment and also well-known as negative prognostic factor in breast cancer malignancies. The new approach using natural chemopreventive substance was developed to inhibit this resistance progress. This study was aimed to investigate whether ethyl acetate fraction of Gynura procumnens (FEG) can prevent chemoresistance through suppressing the MDR1 protein expression. MCF-7 cell was used as chemoresistance cell model. The MCF-7 cells were maintained with 100 nM Dox-contained medium for five weeks. The chemoprevention effect of FEG was investigated by treated MCF-7/Dox with sub-toxic concentration of FEG. The cytotoxic properties of MCF-7 cells were determined using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) assay. Immunofluorescence and western blotting analysis was performed to detect the MDR1 expression. MCF-7/Dox cells need higher concentration for inhibiting cell growth, were compared with MCF-7, shown by IC50value. The MDR1 protein level elevated after Dox exposure in time dependent manner. The FEG treatment decreased MDR-1 protein level with dose dependent manner. FEG in combination with DOX potentiates the DOX effect on breast cancer cell growth inhibition. The FEG prevents the chemoresistance development in breast cancer cell line, MCF-7 induced by Dox through inhibiting MDR1 expression. The additional of FEG enhances Dox effect on cell death induction. Thus, FEG could be developed as co-chemotherapy agent for reverse multidrug resistanc

Efek Hepatoprotektor Ekstrak Etanol Daun Pucuk Merah (Syzygium campanulatum (Korth) dan Daun Cengkeh (Syzygium aromaticum (L.) Merr. & L. M. Perry) pada Tikus Jantan Galur Wistar (Rattus norvegicus) yang diinduksi Paracetamol

Syzygium campanulatum and Syzygium aromaticum contains antioxidant components suchas flavonoids, phenolic, and terpenoids. May have hepatoprotective properties in reducing SGPT and SGOT activity. This research wants to determine the potency of hepatoprotective of ethanolic extract of Syzygium campanulatum (Korth) and Syzygium aromaticum leaf compared with curcuma tablets. This research uses 24 male Wistar rats divided into 6 groups: I, II, III (as a normal, induction, and compared control), group IV, V, VI were treated 105, 210, and 420 mg/kg BW respectively. The study was conducted for 9 days. After 7 days of treatment, treated groups were exposed by hepatotoxic dose of paracetamol (2000 mg/kg BW). The SGPT and SGOT activity of all groups was measured by enzimatic assay. The result can be concluded that Syzygium campanulatum extract was found to be active as hepatoprotective agent with 210 mg/kg BW dosage (SGPT 21.76 ± 3.98 U/L and SGOT 7.32±6.74U/L) as eff ective as with the curcuma tablets (SGPT 23.91 ± 4.41 U/L and SGOT 14.12±5.37 U/L) and the hepatoprotective activity of Syzygium campanulatum extract at a dosage 420 mg/kg BW better than curcuma tablets (SGPT 12.43 ± 6.51 U/L and SGOT 6.64 ± 5.88 U/L). While the hepatoprotec Syzygium campanulatum and Syzygium aromaticum contains antioxidant components such as flavonoids, phenolic, and terpenoids.May have hepatoprotective properties in reducing SGPT and SGOT activity. This research wants to determine the potency of hepatoprotective of ethanolic extract of Syzygium campanulatum (Korth) and Syzygium aromaticum leaf compared with curcuma tablets. This research uses 24 male Wistar rats divided into 6 groups: I, II, III (as a normal, induction, and compared control), group IV, V, VI were treated 105, 210, and 420 mg/kg BW respectively. The study was conducted for 9 days. After 7 days of treatment, treated groups were exposed by hepatotoxic dose of paracetamol (2000 mg/kg BW). The SGPT and SGOT activity of all groups was measured by enzimatic assay. The result can be concluded that Syzygium campanulatum extract was found to be active as hepatoprotective agent with 210 mg/kg BW dosage (SGPT 21.76 ± 3.98 U/L and SGOT 7.32±6.74U/L) as eff ective as with the curcuma tablets (SGPT 23.91 ± 4.41 U/L and SGOT 14.12±5.37 U/L) and the hepatoprotective activity of Syzygium campanulatum extract at a dosage 420 mg/kg BW better than curcuma tablets (SGPT 12.43 ± 6.51 U/L and SGOT 6.64 ± 5.88 U/L). While the hepatoprotective activity of Syzygium aromaticum extracts eff ective as with curcuma tablets at all dosage variation.Daun cengkeh dan daun pucuk merah mengandung komponen antioksidan seperti flavonoid,fenolik, dan terpenoid sehingga diduga memiliki efek hepatoprotektor dalam mengurangi SGPT dan SGOT. Penelitian ini bertujuan untuk mengetahui efek hepatoprotektor dan menentukan potensi hepatoprotektif dari ekstrak etanol daun pucuk merah dan daun cengkeh yang dibandingkan dengan tablet Curcuma. Penelitian ini menggunakan tikus jantan galur wistar yang dibagi menjadi 6 kelompok. Kelompok I, II, III (sebagai kontrol normal, kontrol induksi, dan kontrol pembanding), kelompok IV, V, VI diberi ekstrak uji dengan dosis 105, 210, dan 420 mg/kg BB. Penelitian dilakukan selama 9 hari. Setelah 7 hari diberi perlakuan, semua kelompok diberi parasetamol dosis hepatotoksik kecuali kelompok kontrol normal. Setelah 48 jam diinduksi oleh parasetamol, dilakukan pengukuran SGPT dan SGOT terhadap semua kelompok. Hasil penelitian dapat disimpulkan bahwa ekstrak daun pucuk merah mempunyai aktivitas sebagai hepatoprotektor pada dosis 210 mg/kg BB yang sebanding dengan tablet curcuma (SGPT 21,76±3,98 U/L dan SGOT 7,32±6,74 U/L) dan pada dosis 420 mg/kg BB ekstrak daun pucuk merah memiliki aktivitas hepatoprotektor yang lebih baik dari tablet curcuma (SGPT 12,43±6,51 U/L dan SGOT 6,64±5,88 U/L). Sedangkan ekstrak etanol daun cengkeh mempunyai aktivitas hepatoprotektor yang sebanding dengan tablet curcuma pada dosis 105, 210, dan 420 mg/kgBB

The Cytoprotective and Cell Recovery Properties of Apple Extracts on H2O2 induced-NIH3T3 Cells: An Anti Aging Candidate

Apple contains high concentration of phenolic compounds that protect cells from oxidative stress. The prolong exposure of free radicals may induce cell damage and premature cell aging. Both local and imported apple contain flavonoid, saponin, tannin, steroid, and terpenoid. The extract of local and imported apples showed low toxicity on NIH3T3 fibroblast cells, with IC50 value of 529 and 463 µg/mL, respectively. Both apple extracts (50 – 250 µg /mL) protected three-day-H2O2 induced-cell damage and cell death. Protective effect was observed as the viability increase of treated cells compared to untreated ones. The protective effect of both extracts were higher than the effect of vitamin C as standard antioxidant at this study. Both apple extracts could reverse cell damage caused by three-hour-high concentration H2O2 exposure, similar with vitamin C. Low concentration of both extracts (50 µg /mL) induced the increase of fibroblast cells’ proliferation kinetics. The extract of imported apple showed higher properties of protective, cell recovery and proliferation of fibroblast cells tha local apple, but not statistically significance. This study concludes that the extract of local and imported apples have high potency in cytoprotective effect and cell recovery of damaged cells caused by free radicals induction. Both apple extracts have high potency to be developed the candidate of antiaging and cells’ regeneration agent.Keywords : antiaging, cell recovery, cytoprotective, NIH3T3 cell

Cytotoxic Activity and Apoptosis Induction of Ethanolic Extract of Pericarps of Mangosteen (Garcinia mangostana Linn.) on WiDr Cells and Interaction Study of Alpha-mangosteen to IKK and VEGF Based on Molecular Docking

One of the compounds found efficacious as an anti-proliferative on colon cancer is alpha-mangosteen, a xanthon compound that is abundant in pericarp of mangosteen. This study focused to evaluate anticancer activity of the ethanolic extract of pericarp of mangosteen (Garcinia mangostana Linn.) on WiDr colon cancer cells and to observe the affinity of alpha-mangosteen in inhibiting IKK and VEGF. Cytotoxic effect was tested by MTT assay and apoptosis induction was evaluated by double staining method on WiDr colon cancer cells, while interaction between alpha-mangosteen to the receptors was measured by molecular docking. The ethanolic extract was proven to have cytotoxic activity against WiDr colon cancer cells with IC50 of 25 µg/mL. In addition, the observation of apoptosis induction by double-staining method showed that apoptosis associated the cytotoxic effect of ethanolic extract of pericarp of mangosteen (EPM) on WiDr colon cancer cells. Molecular docking showed that active compounds in pericarp of mangosteen could compete with the native ligand of VEGF because the docking score of alpha-mangosteen was almost equal with native ligand. From these results we could be concluded that the cytotoxic effect of EPM to WiDr cells was mediated by cell apoptosis. This extract was potential to be developed as chemopreventive agent.Keywords : Garcinia mangostana Linn., cytotoxic effect, apoptosis, WiDr cell, molecular docking

EFEKTIVITAS EKSTRAK ETANOL DAUN ADAM HAWA (Rhoeo discolor) DAN DAUN PUCUK MERAH (Syzygium campanulatum Korth.) DALAM MENURUNKAN KADAR GULA DARAH PADA TIKUS PUTIH JANTAN GALUR WISTAR DENGAN PEMBEBANAN GLUKOSA

Penderita diabetes melitus terus semakin meningkat seiring dengan pola hidup yang tidak seimbang. Diabetes melitus merupakan penyakit metabolik yang ditandai dengan peningkatan kadar gula darah di atas normal. Penelitian ini bertujuan untuk menguji efek ekstrak etanol daun adam hawa (Rhoeo discolor) dan daun pucuk merah (Syzygium campanulatum Korth.) terhadap penurunan kadar gula darah pada tikus putih jantan galur wistar yang dibebankan glukosa. Pada penelitian ini tikus jantan galur wistar dibagi menjadi 5 kelompok yaitu kontrol normal, kontrol positif (glibenklamida 0,6 mg/kg bb), kontrol negatif (CMC-Na), tiga kelompok ekstrak etanol daun adam hawa (dosis 100, 200, dan 400 mg/kg bb) dan tiga kelompok ekstrak etanol daun pucuk merah (dosis 300, 600, dan 1200 mg/kg bb. Data diperoleh dengan mengukur kadar gula darah tikus 30 menit setelah pemberian glukosa dan pada menit ke-30, 60, 90, dan 120 setelah perlakuan. Hasil uji penelusuran kandungan senyawa kimia menggunakan Kromatografi Lapis Lipis (KLT) dan pereaksi semprot menunjukkan adanya golongan senyawa flavonoid, alkaloid, dan terpenoid pada ekstrak daun adam hawa, sedangkan ekstrak etanol daun pucuk merah hanya mengandung flavonoid dan terpenoid. Hasil uji statistika menggunakan Anova dengan taraf kepercayaan 95% dan dilanjutkan dengan uji Post Hoc LSD menunjukkan bahwa tidak ada perbedaan yang bermakna antara ekstrak daun adam hawa (dosis 200 dan 400 mg/kg bb) dan ekstrak daun pucuk merah (300 dan 600 mg/kg bb) dengan glibenklamida (0,6 mg/kg bb) dalam aktivitasnya untuk menurunkan kadar glukosa darah tikus. Dosis rendah (100 mg/kg bb) pada adam hawa dan dosis tinggi pada pucuk merah (1200 mg/kg bb) tidak menunjukkan efek hipoglikemik pada tikus. Ekstrak adam hawa dan pucuk merah diduga mempunyai aktivitas antidiabetik yang tergantung dosis (dose dependent). Patients with diabetes mellitus are increasing with the behavior of an unbalanced life. Diabetes mellitus is a metabolic disease characterized by hyperglycemic. This study aims to evaluate the effect of adam hawa (Rhoeo discolor) and pucuk merah (Syzygium campanulatum Korth.) ethanolic leaves extract to decrease blood sugar levels on male wistar rats induced by glucose. Rats were divided into 5 groups: the normal control group, positive control group (glibenclamide 0.6 mg/kg), negative control group (CMC-Na), ethanolic extract of adam hawa group with doses of 100, 200, and 400 mg/kg bw and ethanolic extract pucuk merah group with doses of 300, 600, and 1200 mg/kg bw. Blood glucose levels were measured 30 minutes before and 30, 60, 120 minutes after per oral glucose induction. The results of phytochemical screening using Thin Layer Chromatography (TLC) shown ethanolic extract of adam hawa contained alkaloids, flavonoids, and triterpenoid, while ethanolic extract of pucuk merah only contained flavonoids and terpenoids. Glucose blood levels and AUC datas were statistically analyzed using Oneway Anova and continued with LSD. The datas shown no significant difference between the ethanolic extract of adam hawa (200 and 400 mg/kg bw) and pucuk merah (300 and 600 mg/kg bw) compared with that of glibenclamide (0.6 mg/kg bw) (p>0.05). The ethanolic extract of adam hawa at lower dose (100 mg/kg bw) and pucuk merah at higher dose (1200 mg/kg bw) did not exhibit hipoglicemic effect on rats. Both extracts seems to have antidiabetic properties with dose dependent manner