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PENGARUH EKSTRAK PROPOLIS TERHADAP EKSPRESI CASPASE 3, PROLIFERASI DAN INDUKSI APOPTOSIS PADA SEL KANKER KOLON (CELL LINE WiDr) Anandani, Esti Tantri; Kusnanto, Paulus; Purwanto, Bambang
Biomedika Vol 9, No 2 (2017): Biomedika Agustus 2017
Publisher : Universitas Muhamadiyah Surakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.23917/biomedika.v9i2.5839

Kanker kolorektal mencapai sepuluh persen dari total tipe tumor di seluruh dunia dan merupakan kanker dengan mortalitas tertinggi di seluruh dunia. Kejadian kanker kolon terus meningkat seiring dengan pertambahan penduduk baik di negara berkembang maupun negara maju. Kemoterapi belum memberikan hasil yang optimal dan sering menimbulkan efek samping yang serius karena pada umumnya tidak bekerja spesifik pada sel kanker tetapi juga pada sel normal. Banyak bahan alam yang berpotensial untuk dikembangkan sebagai agen kombinasi seperti propolis, jinten, tapak dara, mengkudu, dan daun dewa. Propolis merupakan suplemen nutrisi yang dihasilkan oleh lebah dan telah digunakan sebagai pengobatan tradisional di dunia. Propolis sudah banyak dijadikan bahan penelitian karena aktivitas antibakteri, antijamur, antivirus dan hepatoprotektifnya. Propolis dan senyawa lainnya telah digunakan untuk mengobati inflamasi, untuk meningkatkan kekebalan tubuh, dan agen anti kanker. Penelitian bertujuan untuk melihat pengaruh pemberian propolis dari daerah Kerjo, Karanganyar, Indonesia terhadap ekspresi caspase 3, proliferasi dan induksi apoptosis pada kultur sel kanker kolon (cell line WiDr).Â Penelitian menggunakan desain eksperimental laboratorik dengan post test with control group design. Penelitian menggunakan kultur sel WiDr (sel kanker kolon) dengan pemberian ekstrak ethanol propolis (EEP). Pengamatan ekspresi caspase 3 dengan metode imunositokimia, sedangkan pengamatan proliferasi dengan metode doubling time dan induksi apoptosis dengan metode double staining. Hasil Penelitian didapatkan EEP dengan IC50 sebesar 140 Âµg/mL meningkatkan ekspresi caspase 3 sebesar 42,1% dibandingkan kelompok kontrol 6,89%. EEP meningkatkan ekspresi caspase 3 sebanding dengan peningkatan konsentrasi EEP. EEP dengan dosis konsentrasi 70, 140, 280 Âµg/mL mampu menekan proliferasi baik pada proliferasi jam ke-24, 48 , maupun 72. EEP dengan IC50 sebesar 140 Âµg/mL meningkatkan induksi apoptosis sebesar 53,16% dibandingkan kelompokkontrol 0,64%. Pemberian EEP meningkatkan induksi apoptosis sebanding dengan peningkatan konsentrasi EEP. Penelitian ini menunjukkan bahwa EEP mempunyai sifat antiproliferasi dan mampu menginduksi apoptosis pada sel WiDr. Â Kata kunci: EEP, caspase 3, proliferasi, apoptosis, WiDr

PENGARUH EKSTRAK ETHANOL PROPOLIS TERHADAP KSPRESI PROTEIN CASPASE 8, APOPTOSIS, DAN PROLIFERASI PADA KULTUR SEL ADENOKARSINOMA KOLON (WiDr) Poernomo, Gigih Rahmadanu; Kusnanto, Paulus; Purwanto, Bambang
Biomedika Vol 10, No 1 (2018): Biomedika Februari 2018
Publisher : Universitas Muhamadiyah Surakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.23917/biomedika.v10i1.5856

Kanker kolorektal merupakan kanker terbanyak ketiga pada pria, kedua pada wanita, dan menjadi penyebab kematian keempat, sekitar 8% dari penyebab kematian karena kanker. Sebagian besar pasien kanker kolorektal didiagnosis dalam stadium yang sudah tidak dapat dioperasi. Propolis diketahui memiliki aktivitas anti kanker. Mekanisme anti kanker propolis melalui induksi apoptosis dan juga melalui penghambatan proliferasi dalam siklus sel. Penelitian ini bertujuan untuk mengkaji efek anti kanker ekstrak ethanol propolis (EEP) yang berasal dari Kerjo, Karanganyar, Indonesia pada kultur sel adenokarsinoma kolorektal (cell line WiDr) melalui pengaruhnya terhadap ekspresi protein Caspase 8, apoptosis dan proliferasi sel. Penelitian ini merupakan penelitian experimental laboratories, post test with control group design. Penelitian dilakukan pada kultur sel WiDr dengan perlakuan pemberian dosis ekstrak ethanol propolis, kontrol positif (5-FU), kombinasi ekstrak ethanol propolis dengan 5-FU, dan kontrol negatif (tanpa obat). Pengamatan ekspresi protein Caspase 8 dilakukan dengan metode imunositokimia, pengamatan apoptosis dilakukan dengan double staining menggunakan pewarnaan akridin oranye–etidium bromide, sedangkan pengamatan proliferasi menggunakan MTT assay dengan doubling time. Hasil Penelitian menunjukkan bahwa EEP cenderung menekan viabilitas sel WiDr dengan IC sebesar 140 µg/mL. EEP konsentrasi 70,140, 280 µg/mL mampu meningkatkan ekspresi protein Caspase 8 dan menginduksi apoptosis yang sebanding dengan peningkatan konsentrasi yang diberikan. EEP konsentrasi 70 µg/mL dapat menghambat proliferasi sel sebanding dengan EEP konsentrasi 140 dan 280 µg/Ml. Penelitian ini menunjukkan EEP mampu menekan viabilitas sel WiDr. Aktivitas ini kemungkinan terkait dengan kemampuannya dalam meningkatkan ekspresi protein Caspase 8 dan apoptosis sebanding dengan peningkatan konsentrasi yang diberikan. EEP pada konsentrasi terkecil yang diuji (70 µg/mL) mampu menghambat proliferasi pada sel WiDr sebanding dengan dosis yang lebih tinggi. Kata kunci: EEP, Caspase 8, apoptosis, proliferasi, cell line WiDr

PENGARUH EKSTRAK ETANOL PROPOLIS TERHADAP EKSPRESI PROTEIN BCL2 DAN P21 DALAM MENEKAN PROLIFERASI DAN MENGINDUKSI APOPTOSIS PADA KULTUR SEL HEPATOMA (Hep G2) Salimah, Kun; Kusnanto, Paulus; Purwanto, Bambang
Biomedika Vol 11, No 1 (2019): Biomedika Februari 2019
Publisher : Universitas Muhamadiyah Surakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.23917/biomedika.v11i1.5364

Hepatocellular carcinoma (KHS) is 90% of primary tumors in the liver. One of natural treatment that have anti-cancer activity is propolis. Its anti-cancer activity works through induction of apoptosis and inhibition of cell proliferation. Bcl2 belongs to the family group B-cell lymphoma 2 protein which acts as an intrinsic pathway apoptotic protein. p21 is a tumor suppressor protein that has a primary function in inhibiting cell cycle progression. This study was aimed to determine the anti-cancer effect of propolis ethanol extract (EEP) originating from Kerjo, Karanganyar on hepatocellular cancer cell culture (Hep G2) through its potential as an anticancer in increasing p21 protein expression, reducing bcl2 protein expression in suppressing apoptosis and reducing cell proliferation . This research is an experimental laboratories with post test with control group design. Performed on Hep G2 cells with the treatment of EEP concentrations (½IC50, IC50, 2IC50), sorafenib IC50, a combination of EEP IC50 + sorafenib IC50, and controls. Observation of Bcl2 and p21 protein expression by immunocytochemistry method, observing cell proliferation with doubling time, observing apoptosis with flowcytometry. We use ANOVA statistical test, followed by the Tuckey post hoc test, were stated to be significant if p< 0.05. The inhibition concentration of 50% Hep G2 cells by EEP was 65.2 μg / mL, and sorafenib (6.08 μg / mL). EEP concentration of 2IC50 (p = 0.001), sorafenib 1 IC50 (p = 0.001), 1IC50 EEP + 1IC50 sorafenib (p= 0.001) able to increase p21 protein expression. The 2 EEP concentration (p= 0.040), was able to reduce the expression of Bcl2 protein, with p= 0.001. The strength of EEP 1IC50 is equivalent to sorafenib IC50 in suppressing 24-hour proliferation (p> 0.05). The strength of EEP ½IC50 is equivalent to sorafenib IC50 in suppressing proliferation 48 hours (p> 0.05). The strength of EEP 1IC50 is better than sorafenib IC50 in suppressing proliferation of 72 hours (p> 0.05). All EEP concentrations increase p21 protein expression, with the EEP 2IC50 group providing the strongest effect. All EEP concentrations were able to reduce cell proliferation, with the EEP 2IC50 group, giving the strongest effect. It was concluded that concentration of 2IC50 EEP has an effect on decreasing the expression of bcl2 protein and hepatoma cell apoptosis.Keywords: Ethanol Extract Propolis, Bcl2, P21, Apoptosis, Proliferation, Hep G2 Cells

The Effect of Simvastatin on Nitric Oxide and hs-CRP Levels in patients with Liver Cirrhosis at Dr. Moewardi Hospital, Surakarta Hariyanto, Budhi; Kusnanto, Paulus; Purwanto, Bambang
Indonesian Journal of Medicine Vol 4, No 1 (2019)
Publisher : Masters Program in Public Health, Universitas Sebelas Maret, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (193.661 KB)

Background: The prevalence of liver cirrhosis (SH) worldwide is at seventh place as the cause of death. Increased pressure in the portal vein Portal Hypertension (HP) causes esophageal varices and the more likely the patient to bleed. Endothelial dysfunction in cirrhosis of the liver causes a decrease in nitric oxide (NO) levels resulting in an imbalance between relaxation factors and portal vein contractions and increasing hs-CRP levels. Simvastatin can increase NO levels and improve endothelial dysfunction in patients with SH and can reduce levels of hs-CRP. This study aimed to determine the effect of simvastatin administration on nitric oxide and hs-CRP level in patients with liver cirrhosis.Subjects and Method: This was a randomized controlled trial conducted at Dr. Moewardi hospital, Surakarta, Central Java. A sample of 29 patients was divided into treatment and control groups. 14 patients in the treatment group were given simvastatin dose of 20 mg. 15 patients in the control group received a placebo. The dependent variable was a nitric oxide and hs-CRP levels. The independent variable was Simvastatin administration. Nitric oxide serum level was measured by an enzyme-linked immunosorbent assay (ELISA). Hs-CRP level was measured by the immunechemiluminescent method. The data were analyzed by t-test.Results: NO level after Simvastatin 20 mg administration (Mean= 7.93 µg/mL; SD= 1.78 µg/ mL) was higher than before (Mean= 5.14 µg/mL; SD= 2.25 µg/mL) and it was statistically significant (p = 0.001). Hs-CRP level after Simvastatin 20 mg administration (Mean= 2.04 mg/L; SD= 0.80 mg/L) was lowered than before (Mean= 7.82 mg/L; SD= 1.15 mg/L) and it was statistically significant (p= 0.001).Conclusion: Simvastatin administration increases NO serum levels and reduces the hs-CRP level in patients with liver cirrhosis.Keywords: nitric oxide, hs-CRP, simvastatin, liver cirrhosisCorrespondence: Budhi Hariyanto. Masters Program in Family Medicine, Universitas Sebelas Maret. Jl. Ir Sutami 36A, Surakarta 57126, Centra Java. Email: antohariyanto787@gmail.com. Mobile: 082328010442.Indonesian Journal of Medicine (2019), 4(1): 9-14https://doi.org/10.26911/theijmed.2019.04.01.02

PENGARUH EKSTRAK PROPOLIS TERHADAP EKSPRESI CASPASE 3, PROLIFERASI DAN INDUKSI APOPTOSIS PADA SEL KANKER KOLON (CELL LINE WiDr) Esti Tantri Anandani; Paulus Kusnanto; Bambang Purwanto
Biomedika Vol 9, No 2 (2017): Biomedika Agustus 2017
Publisher : Universitas Muhamadiyah Surakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.23917/biomedika.v9i2.5839

Kanker kolorektal mencapai sepuluh persen dari total tipe tumor di seluruh dunia dan merupakan kanker dengan mortalitas tertinggi di seluruh dunia. Kejadian kanker kolon terus meningkat seiring dengan pertambahan penduduk baik di negara berkembang maupun negara maju. Kemoterapi belum memberikan hasil yang optimal dan sering menimbulkan efek samping yang serius karena pada umumnya tidak bekerja spesifik pada sel kanker tetapi juga pada sel normal. Banyak bahan alam yang berpotensial untuk dikembangkan sebagai agen kombinasi seperti propolis, jinten, tapak dara, mengkudu, dan daun dewa. Propolis merupakan suplemen nutrisi yang dihasilkan oleh lebah dan telah digunakan sebagai pengobatan tradisional di dunia. Propolis sudah banyak dijadikan bahan penelitian karena aktivitas antibakteri, antijamur, antivirus dan hepatoprotektifnya. Propolis dan senyawa lainnya telah digunakan untuk mengobati inflamasi, untuk meningkatkan kekebalan tubuh, dan agen anti kanker. Penelitian bertujuan untuk melihat pengaruh pemberian propolis dari daerah Kerjo, Karanganyar, Indonesia terhadap ekspresi caspase 3, proliferasi dan induksi apoptosis pada kultur sel kanker kolon (cell line WiDr). Penelitian menggunakan desain eksperimental laboratorik dengan post test with control group design. Penelitian menggunakan kultur sel WiDr (sel kanker kolon) dengan pemberian ekstrak ethanol propolis (EEP). Pengamatan ekspresi caspase 3 dengan metode imunositokimia, sedangkan pengamatan proliferasi dengan metode doubling time dan induksi apoptosis dengan metode double staining. Hasil Penelitian didapatkan EEP dengan IC50 sebesar 140 µg/mL meningkatkan ekspresi caspase 3 sebesar 42,1% dibandingkan kelompok kontrol 6,89%. EEP meningkatkan ekspresi caspase 3 sebanding dengan peningkatan konsentrasi EEP. EEP dengan dosis konsentrasi 70, 140, 280 µg/mL mampu menekan proliferasi baik pada proliferasi jam ke-24, 48 , maupun 72. EEP dengan IC50 sebesar 140 µg/mL meningkatkan induksi apoptosis sebesar 53,16% dibandingkan kelompokkontrol 0,64%. Pemberian EEP meningkatkan induksi apoptosis sebanding dengan peningkatan konsentrasi EEP. Penelitian ini menunjukkan bahwa EEP mempunyai sifat antiproliferasi dan mampu menginduksi apoptosis pada sel WiDr. Kata kunci: EEP, caspase 3, proliferasi, apoptosis, WiDr

Non-endoscopic Examination as Predictor of Varices Degree in Liver Cirrhosis Patients Who have Experienced Esophageal Variceal Bleeding Paulus Kusnanto; Marcellus Simadibrata; Irsan Hasan
The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy VOLUME 12, NUMBER 1, April 2011
Publisher : The Indonesian Society for Digestive Endoscopy

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24871/121201115-22

Background: Standard diagnosis for determining the degree of varices is by endoscopy. However, sometimes there are obstacles in the implementation of endoscopy. Based on the factors, we need to know the parameters of non-endoscopic examination which include ascites, splenomegaly, thrombocytopenia, Child-Pugh, portal vein diameter as a predictor of the degree of liver cirrhosis patients with varices who have experienced esophageal variceal bleeding. Method: The study design was cross-sectional study. The study was conducted on hospitalized patients in Cipto Mangunkusumo hospital, Gatot Subroto hospital, and Kraton hospital from September 2008 to November 2009. The patients were liver cirrhosis patients with history of upper gastrointestinal bleeding, no present bleeding, and hemodynamically stable. Examination of predictor factors in the patients such as ascites, splenomegaly, thrombocytopenia, Child-Pugh and portal vein diameter were done. Statistical analysis was performed with student’s t-test, Mann-Whitney test, and stepwise multivariable logistic regression. Results: The study involved 44 patients with liver cirrhosis who have esophageal variceal bleeding. Based on the results of endoscopic examination, large varices (F3) were found in 21 (47.73%) patients, small varices (F1 F2) in 23 (52.27%) patients, located on the distal esophagus extending to the medial (86.4%), with red color sign present (54.5%). Results of non-endoscopic examination such as splenomegaly, ascites, thrombocytopenia, portal vein diameter and Child-Pugh score was known not to be associated with the degree of esophageal varices (p 0.05). Conclusion: Non-endoscopic examination was not related to the degree of varices in liver cirrhosis patients who have experienced esophageal variceal bleeding. Keywords: esophageal variceal bleeding, liver cirrhosis, predictor factors, endoscopic criteria

Histopathology of Helicobacter pylori in Chronic Dyspepsia Patients Wildan Nur; Paulus Kusnanto; Triyanta Yuli Pramana; Michael Tantoro Harnomo; Oyong Oyong; Ambar Mudigdo
The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy VOLUME 13, NUMBER 3, Desember 2012
Publisher : The Indonesian Society for Digestive Endoscopy

ABSTRACTBackground: Helicobacter pylori (H. pylori) are the most common infection found in dyspepsia cases. This infection is almost always found in digestive tract inflammation and commonly develops into chronic gastritis. Meanwhile, chronic gastritis is a condition assumed as the early event in pathological abnormalities of the stomach which finally may develop into carcinoma of the gaster. In Indonesia, data describing the incidence of H. pylori infection based on the histopathological appearance, location of specimen collection, inflammatory degree, and age of chronic dyspepsia patients is not yet available. The aim of this study is to determine the incidence of H. pylori based on histopathology appearance in chronic dyspepsia patients in Moewardi Hospital Surakarta.Method: This study is a cross sectional descriptive study by performing endoscopy-biopsy and histopathology examination to chronic dyspepsia patients who came to Gastroenterohepatology Clinic, Department of Internal Medicine, Moewardi Hospital, Surakarta on 1 January 2009 - 31 December 2010.Results: More than 90% subjects were 40 year old with the distribution of majority patients were 46-55 year old (32.43%). The most commonly found endoscopic appearance in subjects with positive H. pylori was superficial chronic gastritis (81.08%) with mild inflammatory degree (64.86%) and majority located in the antrum 97.3%.Conclusion: The proporsion of H. pylori infection in male and female was almost equal and was mostly found in the age group of 46-55 year old. This infection frequently happens in chronic dyspepsia who has histopathologic appearance of superficial chronic gastritis with mild inflammatory degree in the antrum area. Keywords: Helicobacter pylori, superficial chronic gastritis, age

A 17-Years Old Man of Colitis Tuberculosis with Fistula Perianal Paulus Kusnanto; Marcellus Simadibrata; Ari Fahrial Syam; Achmad Fauzi; Murdani Abdullah; Dadang Makmun; Chudahman Manan; Daldiyono Daldiyono; Abdul Aziz Rani; Ening Krisnuhoni
The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy VOLUME 9, ISSUE 3, December 2008
Publisher : The Indonesian Society for Digestive Endoscopy

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24871/932008103-106

Tuberculosis (TB) remains to be one of the most common problems in developing countries such as Indonesia. It can involve many organs including gastrointestinal tracts. Colonic tuberculosis is an ancient disease and has long been recognized. However, it is sometimes difficult to make early diagnosis due to its nonspecific signs and symptoms. Perianal granulomas or perianal fistula presents some degree of diagnostic difficulty. A spectrum of diseases can produce granulomas in perianal region and perineum. Most are infectious or inflammatory diseases. Standard histological diagnosis often less significant to clarify the etiology and treatment will vary from one to another disease entity. In this report, we present a case 17-years old man with colitis TB and fistula perianal, in which the initial diagnostic workup suggested Crohn’s disease. Mantoux test, Acid Fast Bacilli test on fecal examination and polymerase chain reaction analysis revealed negative result. The chest X-ray was normal; while the fistulography X-ray: revealed 2 orifices in rectosigmoid area. The colonoscopy revealed mucosa edema with ulceration in rectosigmoid and pedincular polyp in the caecum. Initial colonoscopy diagnosis was Crohn’s disease with differential diagnosis colitis ulcerative, colitis TB, colitis infection. results of direct or post homogenizes examination (Ziel–Nielsen staining) revealed that no acid-fast bacilli was found. Multiple biopsies were done, which indicated mucosa edema with ulceration in rectosigmoid area, pedincular polyp in the caecum, and surrounded by fistula perianal; while histopathological examination showed inflammatory-caseating-epithelioid-granulomas and giant cells (Langhans datia cell) caused by tuberculosis. It highlights the need for awareness of intestinal TB along with the differential diagnosis of chronic intestinal disease. Standard regimen of antituberculosis treatment was given and the patient showed good clinical response. Keywords: Crohn’s disease, caseating epithelioid granuloma, giant cell, colitis TB, perianal fistula

Phenotype and Conditioning Medium on Umbilical Cord-Mesenchymal Stem Cell (UC-MSC) Kusnanto, Paulus; Purwanto, Bambang; Wasita, Brian; Widyaningsih, Vitri
The Indonesian Journal of Gastroenterology, Hepatology, and Digestive Endoscopy Vol 24, No 2 (2023): VOLUME 24, NUMBER 2, August, 2023
Publisher : The Indonesian Society for Digestive Endoscopy

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24871/2422023127-131

Stem cells are unspecialized cells that differentiate into more specialized cells for various tissues in the human body. Stem cells have the characteristics and ability to become themselves by performing various mitoses and differentiating different cells. Stem cells also have great potential for tissue regeneration. Detection and isolation of specific cells is very important in understanding biological processes such as development, aging, regeneration, and cell pathogenesis. Therefore, stem cells are present as an invaluable therapeutic option. Before stem cells can be applied, it is important to know the characteristics of the specific function cells or phenotypic markers. Isolation and characteristics of stem cells on culture morphology or immunotaining on specific markers. Immunohistochemistry can provide a general description of the expression of markers in a cell under certain conditions. Surface markers are the basis for identifying the characteristics and isolation of stem cells. The basic characteristics of stem cells are one that influences the outcome of the therapy to be carried out. The presence of marker cell surface expressions can also be a differentiator in searching for the origin of the stem cells used6. MSC culture method was performed with DMEM + 5% HPL. UC-MSCs with phenotype differentiation were found to be CD73+, CD90+, CD105+, Lin- while the parameters found were IL-6, SDF-1, BDNF, FGF, NGF, IL-12p70, IL-23, IL-18, IL-10 , VEGF, EGF, IL-17A, IFN-, IL-1β, PDGF, MCP-1.The results obtained are in accordance with the standard content of UC-MSC cells which can be continued as cell research.

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