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A Model of Rat Thrombocytopenia Induced by Cyclophosphamide Kristiana, Hery; Nailufar, Florensia; Winoto, Imelda L; Tjandrawinata, Raymond R
JFIOnline | Print ISSN 1412-1107 | e-ISSN 2355-696X Vol 6, No 3 (2013)
Publisher : Indonesian Research Gateway

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (288.566 KB)

This study is aimed at optimizing the dose of cyclophosphamide in thrombocytopenia rat model. A suitable drug dosage that coulds induce thrombocytopenia rat model. A suitable drug dosage that could induce thrombocytopenia in rat was then investigated. Animal model of 25 rats were randomly divided into five groups: Normal (group A), Cyclophosphamide low dose (group B), medium dose (group C), high dose (group D), very high dose (group E) with each of 5 rats. The four dosage groups were given 25, 50, 100, 150 mg/kg BW cyclophosphamide, respectively. Cyclophosphamide was given by subcutaneous injection once a day for 3 consecutive days. All groups were under investigation for 8 days. The result suggested that a decrease in the platelet count, white blood cell, and mean corpuscular volume of all group cyclophosphamide induced at the 7th day were significantly different than that of Normal. The platelets count was reduced but fluctuated greatly, all of the rats dies in group E at 7th day and group D at 8th day. Though all 4 dosages successfully initiated thrombocytopenia as the platelets number dorpped at the 7th day, the low dose was considered to be a suitable one that was of high efficacy and low toxicity. Thus, using Wistar rats challenged by subcutaneous injection of cyclophosphamide 25 mg/kg per day for 3 consecutive days showed one simple, feasible and stable rat thrombocytopenia model that could be used for phamacodynamic test of the drugs which are supposed to have platelet increasing effect. Keywords : rat, thrombocytopenia, cyclophosphamide

A Model of Rat Thrombocytopenia Induced by Cyclophosphamide Kristiana, Hery; Nailufar, Florensia; Winoto, Imelda L; Tjandrawinata, Raymond R
Jurnal Farmasi Indonesia Vol 6, No 3 (2013)
Publisher : Jurnal Farmasi Indonesia

This study is aimed at optimizing the dose of cyclophosphamide in thrombocytopenia rat model. A suitable drug dosage that coulds induce thrombocytopenia rat model. A suitable drug dosage that could induce thrombocytopenia in rat was then investigated. Animal model of 25 rats were randomly divided into five groups: Normal (group A), Cyclophosphamide low dose (group B), medium dose (group C), high dose (group D), very high dose (group E) with each of 5 rats. The four dosage groups were given 25, 50, 100, 150 mg/kg BW cyclophosphamide, respectively.Â Cyclophosphamide was given by subcutaneous injection once a day for 3 consecutive days. All groups were under investigation for 8 days. The result suggested that a decrease in the platelet count, white blood cell, and mean corpuscular volume of all groupÂ cyclophosphamide induced at the 7th day were significantly different than that of Normal. The platelets count was reduced but fluctuated greatly, all of the rats dies in group E at 7th day and group D at 8th day. Though all 4 dosages successfully initiated thrombocytopenia as the platelets number dorpped at the 7th day, the low dose was considered to be a suitable one that was of high efficacy and low toxicity. Thus, using Wistar rats challenged by subcutaneous injection ofÂ cyclophosphamide 25 mg/kg per day for 3 consecutive days showed one simple, feasible and stable rat thrombocytopenia model that could be used for phamacodynamic test of the drugs which are supposed to have platelet increasing effect. Keywords : rat, thrombocytopenia,Â cyclophosphamide

Peptides Hydrolysate Derived from Collagen of Snakehead Murrel (Channa striata) Skin Demonstrate Anti-cholesterol and Anti-oxidant activities Sinaga, Wenny Silvia L. Br.; Ismaya, Wangsa T.; Retroningrum, Debbie S.; Tjandrawinata, Raymond R.; Suhartono, Maggy T.
HAYATI Journal of Biosciences Vol. 27 No. 2 (2020): April 2020
Publisher : Bogor Agricultural University, Indonesia

Anti-cholesterol and anti-oxidant play a crucial role to combat cardiovascular disease (CVD), due to formation of arterial plagues from oxidation of cholesterol. In the past decades, bioactive peptides demonstrating anti-cholesterol and anti-oxidant activities have emerged as the alternative drugs. In this study, acid soluble collagen was extracted from the skin of snakehead murrel and employed to induce secretion of collagenase by Bacillus licheniformis F11.4. The collagenases secreted were in turn used to produce peptides hydrolysate and were grouped in two distinct collagenase fractions, designated as fraction D and F. Peptides hydrolysate produced by the fraction D was found to demonstrate HMG-CoA inhibitor activity comparable to pravastatin and limited anti-oxidant activity. Meanwhile, peptides hydrolysate generated using the fraction F demonstrated anti-oxidant activity comparable to BHT (2mM), vitamin C (2mM), and vitamin E (2mM), but limited HMG-CoA activity. Combination of the fraction D and F resulted in substantial HMG-CoA inhibition and anti-oxidant activities.

Food Origin Fibrinolytic Enzyme With Multiple Actions Laurentia Stephani; Raymond Rubianto Tjandrawinata; Diana Nur Afifah; Yanti Lim; Wangsa Tirta Ismaya; Maggy Thenawidjaja Suhartono
HAYATI Journal of Biosciences Vol. 24 No. 3 (2017): July 2017
Publisher : Bogor Agricultural University, Indonesia

Many health related problems such as cardiovascular diseases are associated with the formation of excessive clot in the blood (thrombus). Approaches in cardiovascular disease treatment are preventing the formation or removing the thrombus. The present thrombolytic agents can be classified as plasminogen activators, fibrinolytic enzyme which directly degrades fibrinogen or fibrin and heparin type which act as thrombin inhibitor. Recently, microbial fibrinolytic enzymes of food origin receive more attention that leads to escalating efforts to explore traditional fermented foods as the natural sources. We have successfully isolated microorganism from Indonesian fermented soybean tofu dregs “Oncom” that secretes fibrinolytic enzyme. The microorganism identified as Stenotrophomonas sp. is unique because most of the reported fibrinolytic microorganism belongs to Bacillus sp. This isolate was found to produce extracellular fibrinolytic enzyme which could degrade fibrinogen and fibrin directly as determined by fibrinogen zymography and fibrin plate methods. More importantly, the 30-kD purified enzymes was found to demonstrate not only fibrin and fibrinogen degradation capabilities, but also acted as thrombin inhibitor as determined using specific substrates for thrombin. This is the first report of a fibrinolytic enzyme that demonstrates additional synergistic activities. This finding accentuates the importance of further development of the enzyme into a powerful agent to treat the thrombus-related disease effectively.

Peptides Hydrolysate Derived from Collagen of Snakehead Murrel (Channa striata) Skin Demonstrate Anti-cholesterol and Anti-oxidant activities Wenny Silvia L. Br. Sinaga; Wangsa T. Ismaya; Debbie S. Retroningrum; Raymond R. Tjandrawinata; Maggy T. Suhartono
HAYATI Journal of Biosciences Vol. 27 No. 2 (2020): April 2020
Publisher : Bogor Agricultural University, Indonesia

Anti-cholesterol and anti-oxidant play a crucial role to combat cardiovascular disease (CVD), due to formation of arterial plagues from oxidation of cholesterol. In the past decades, bioactive peptides demonstrating anti-cholesterol and anti-oxidant activities have emerged as the alternative drugs. In this study, acid soluble collagen was extracted from the skin of snakehead murrel and employed to induce secretion of collagenase by Bacillus licheniformis F11.4. The collagenases secreted were in turn used to produce peptides hydrolysate and were grouped in two distinct collagenase fractions, designated as fraction D and F. Peptides hydrolysate produced by the fraction D was found to demonstrate HMG-CoA inhibitor activity comparable to pravastatin and limited anti-oxidant activity. Meanwhile, peptides hydrolysate generated using the fraction F demonstrated anti-oxidant activity comparable to BHT (2mM), vitamin C (2mM), and vitamin E (2mM), but limited HMG-CoA activity. Combination of the fraction D and F resulted in substantial HMG-CoA inhibition and anti-oxidant activities.

Overproduction, Purification and Refolding of codon-optimized Hepatitis B Virus X Protein Subgenotype B3 in Escherichia coli BL21(DE3) Anita Artarini; Armini Syamsidi; Anindyajati Anindyajati; Raymond R. Tjandrawinata; Debbie S. Retnoningrum
HAYATI Journal of Biosciences Vol. 29 No. 2 (2022): March 2022
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.29.2.164-170

Hepatitis B virus (HBV) infects human and causes chronic liver infection, leading to liver cirrhosis and hepatocellular carcinoma. HBV X (Hbx) protein is known to interact with tumor suppressor protein p53 and block its translocation into the nucleus. This study outlines the overproduction of Hbx protein from HBV subgenotype B3 in Escherichia coli BL21(DE3), including its purification and refolding. The gene encoding Hbx was first codon-optimized and inserted into pET16b. The recombinant plasmid was then transformed into E. coli BL21(DE3) as an expression host. Optimization of Hbx expression was performed with variation of IPTG concentration and overproduction temperature. The results showed that Hbx protein was optimally induced by 0.075 mM IPTG and overproduction of Hbx at 17, 25, and 37°C exhibited no difference in protein level and location. The optimal refolding of Hbx was obtained using 0.1 M arginine prior to elution from Nickel column using 100 mM imidazole and 0.25 M arginine. Hbx migrates differently in SDS-PAGE reducing and non-reducing, while the melting curve pattern in TSA analysis changed after the refolding step. Essentially, this purified Hbx protein could potentially be used for interaction study with p53 and the inhibitor candidate of the protein.

A Simple Liquid-Liquid Fractionation (LLF) Method for Isolating Deoxyandrographolide dan Andrographolide from Herbs of Andrographis paniculata (Burm., F) Ness and Its Cytotoxic Activity on 3T3-L1 Preadipocyte Cells Putri Rachma Novitasari; Novia Tri Astuti; Suwijiyo Pramono; Raymond Tjandrawinata; Agung Endro Nugroho
Journal of Food and Pharmaceutical Sciences Vol 8, No 3 (2020): J.Food.Pharm.Sci
Publisher : Institute for Halal Industry and System (IHIS) Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jfps.875

The main bitter constituents of sambiloto (Androgaphis paniculata (Burm., F) Ness) are diterpene lactones, namely andrographolide and deoxyandrographolide which have been reported to have antidiabetic, cytotoxic, antiatherosclerosis, anti-inflammatory, and antimicrobial activity. There are many studies that performed the isolation of deoxyandrographolide and andrographolide from A. paniculata herbs, but most of them included several steps that make them not efficient. This research was conducted to do an isolation of deoxyandrographolide and andrographolide through liquid-liquid fractionation (LLF) due to its simplicity, low cost, and time efficient. The extraction of deoxyandrographolide and andrographolide from the herbs was carried out using chloroform as the solvent by using Soxhlet apparatus, and LLF was performed to isolate the compounds. The identities of the compounds were confirmed by TLC scanner compared to its standard references. Hence, these present methods were successfully isolated and determined deoxyandrographolide and andrographolide of A. paniculata. The compounds were also showed relatively moderate cytotoxicity on 3T3-L1 cell lines using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay, with LC50 of deoxyandrographolide and andrographolide; 29.3173 µg/mL and 37.7011 µg/mL, respectively.

Docking Molekular dari Trigonella foenum-graceum sebagai Antidiabetes menggunakan Molegro Virtual Docking Andri Prasetiyo; Esti Mumpuni; Raymond R. Tjandrawinata
Jurnal Jamu Indonesia Vol. 4 No. 2 (2019): Jurnal Jamu Indonesia
Publisher : Pusat Studi Biofarmaka Tropika LPPM IPB; Tropical Biopharmaca Research Center - Bogor Agricultural University

Trigonella foenum-graceum atau fenugreek digunakan secara luas sebagai obat tradisional untuk pengobatan diabetes tetapi mekanisme kerjanya masih belum jelas. Penelitian bertujuan memprediksi senyawa dalam fenugreek yang berkhasiat sebagai antidiabetes secara in-silico dengan menggunakan perangkat lunak Molegro Virtual Docking . Docking dilakukan 10 Senyawa uji dalam fenugreek yaitu 4-hidroxyisoleucine, coumarine, diosgenin, galactomannan, isovitexin, quarcetin, tigogenin, trigoneline, vitexin dan yamogenin dengan 3 reseptor yaitu sugar beet alpha-glucosidase- (PDB ID : 3W37), human dipeptidyl peptidase-4 (PDB ID : 1X70), human peroxisome proliferator activated gamma (PDB: 2PRG) serta senyawa pembanding acarbose, sitagliptin dan rosiglitazone. Dari 10 senyawa uji, galactomanann memiliki nilai Rerank Score/RS paling rendah di dua reseptor yaitu alpha glucosidase dan peroxisome proliferator activated gamma dengan nilai berturut turut -116.56 kcal/mol dan -131.18 kcal/mol dan nilai RS acarbose -113.60 kcal/mol dan rosiglitazone -124.54 kcal/mol . Dari 10 senyawa uji, tigogenin memiliki nilai RS paling rendah direseptor dipeptidyl peptidase-4 dengan nilai RS -86.54 kcal/mol dan nilai RS sitagliptin -87.02 kcal/mol. Berdasarkan nilai RS, galactomannan diprediksi memiliki aktivitas antidiabetes yang bekerja pada reseptor alpha-glucosidase dan peroxisome proliferator activated gamma sedangkan tigogenin diprediksi memiliki aktivitas antidiabetes yang bekerja pada reseptor dipeptidyl peptidase-4.

Mekanisme Molekuler Imunomodulasi oleh Huangqi (Astragali Radix) Menggunakan Sistem Farmakologi Jejaring Raymond Rubianto Tjandrawinata; Anindini Winda Amalia; Hartati Tuna; Viedya Novalinda Said; Santi Tan
JURNAL ILMU KEFARMASIAN INDONESIA Vol 20 No 2 (2022): JIFI
Publisher : Fakultas Farmasi Universitas Pancasila

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35814/jifi.v20i2.1301

Astragali Radix (Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao, also known as Huangqi) has been used to improve stamina and immunity for thousands of years. However, the bioactive components and their mechanisms are still unclear, limiting its clinical application. In this research, for the first time, network pharmacology was used to investigate the molecular mechanisms of Huangqi and its bioactive components that play an important role in improving the immune system. An amount of 1,430 immunomodulation-related target proteins was collected from the human genome database, with 171 target proteins from 20 bioactive components, and resulted in 60 intersection target proteins. Subsequently, the networks of common target and protein-protein interaction (PPI) were analyzed. The results showed that Huangqi influenced the modulations of lymphocytes (B cells and T cells), natural killer cells, macrophages, cytokines, immunoglobulins, and immune signal transduction. To a certain limit, this study revealed the potential bioactive components and pharmacological mechanisms of Huangqi in immunomodulation. It was able to direct novel drug development for treating immune system deficiency.

Anti‐diabetic effect of andrographolide from Sambiloto herbs (Andrographis paniculata (Burm.f.) Nees) through the expression of PPARγ and GLUT‐4 in adipocytes Novia Tri Astuti; Putri Rachma Novitasari; Raymond Tjandrawinata; Agung Endro Nugroho; Suwijiyo Pramono
Indonesian Journal of Biotechnology Vol 27, No 4 (2022)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijbiotech.68800

Andrographolide has been shown to have a pharmacological effect as an antidiabetic. Nevertheless, the comprehensive mechanism of action has yet to be determined. Andrographolide is a primary component of the sambiloto herb (Andrographis paniculata (Burm.f.) Nees), in which a simple isolation process can obtain high yields. This study aimed to explain the anti‐diabetic effect of andrographolide compared to pioglitazone (a positive control) on glucose uptake by measuring the expression levels of peroxisome proliferator‐activated receptor gamma (PPARγ) and glucose transporter type 4 (GLUT‐4) genes in 3T3‐LI mouse adipocytes as an in vitro model. The differentiation of mature adipocytes from 3T3‐L1 fibroblasts was induced with 3‐isobutyl‐1‐methylxanthine, dexamethasone, and insulin. Andrographolide was provided through direct isolation from A. paniculata herbs. The gene expression was detected using the reverse transcription‐polymerase chain reaction (RT‐PCR). Pioglitazone and andrographolide significantly increased glucose uptake capability. Andrographolide was able to increase the mRNA levels of PPARγ and GLUT‐4 compared to pioglitazone with the best concentration at 5.6 µM. In conclusion, andrographolide can improve glucose uptake by increasing mRNA levels of PPARγ and GLUT‐4 that encodes protein, which are key factors for glucose homeostasis. Therefore, this finding further establishes the potency of andrographolide from A. paniculata as an antidiabetic.

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