EFFECT OF STATINS ON ENDOTHELIAL PROGENITOR CELL (EPC) MIGRATION FROM PERIPHERAL BLOOD OF STABLE CORONARY ARTERY DISEASE PATIENT

Penelitian Klinis ABSTRAK   Efek Pemberian Statin Terhadap Migrasi Endothelial Progenitor Cell (EPC) Pada Darah Tepi Penderita Penyakit Jantung Koroner Stabil Tyagita Verdena Rani Savitri, Yudi Her Oktaviono, Djoko Soemantri   Latar Belakang: Endothlelial progenitor cell (EPC) berpartisipasi dalam perbaikan endotel dan pertumbuhan pembuluh darah baru. Farmakoterapi kardiovaskular telah dibuktikan dapat memperbaiki jumlah dan fungsi EPC pada penderita dengan risiko kardiovaskular. Banyak studi melaporkan bahwa statin memiliki efek yang menguntungkan terhadap EPC dengan meningkatkan jumlah dan fungsi EPC, termasuk didalamnya adalah fungsi migrasi. Oleh karena itu, kami melakukan penelitian untuk menganalisis efek tiga statin yang berbeda terhadap migasi EPC. Tujuan: Untuk menganalisis efek pemberian statin terhadap migrasi EPC pada darah tepi penderita penyakit jantung koroner stabil. Metode: Penelitian ini kami lakukan secara in vitro true experimental post-test only control group design. Sel mononuklear diisolasi dari darah tepi penderita penyakit jantung koroner stabil dan dilakukan kultur dalam medium Stemline II Hematopoietic Stemcell Expansion Medium selama 3 hari. Kemudian sampel dibagi menjadi empat kelompok yaitu kelompok simvastatin 0.5 µmol/L,, atorvastatin 0.5 µmol/L, rosuvastatin 0.5 µmol/L dan kelompok kontrol kemudian diinkubasi selama 48 jam. Metode imunositokimia dilakukan untuk identifikasi EPC dengan mengevaluasi ekspresi CD34+. Pada hari ke-5 kultur, sel di pindahkan ke bagian atas transwell system sebanyak 5x105 sel per sumur perlakuan , kemudian di inkubasi selama 1 hari. Sel yang berpindah pada sumur transwell system bagian bawah dihitung dengan automatic cell counter dengan pewarnaan typhan blue. Data dianalisis dengan independent t test dan ANOVA. Hasil: Hasil independent t test terhadap hasil migrasi pada transwell system menunjukkan peningkatan bermakna terhadap migrasi EPC pada kelompok simvastatin, atorvastatin, dan rosuvastatin dibandingkan dengn kelompok kontrol (234000 ± 1290. 994, 265000 ± 1290. 994, 203000 ± 1290. 994 vs 174071.43 ± 1426. 785, p<0.05). Migrasi EPC juga berbeda antar kelompok statin, dimana efek tertinggi didapatkan pada kelompok atorvastatin. Migrasi EPC pada kelompok atorvastatin lebih tinggi daripada kelompok simvastatin (265000 ± 1290. 994 vs 234000 ± 1290. 994, p<0.05), dan simvastatin lebih tinggi daripada kelompok rosuvastatin (234000 ± 1290. 994 vs 203000 ± 1290. 994, p<0.05). Pemeriksaan imunositokimia menunjukkan ekspresi positif terhadap CD34+. Kesimpulan: Statin meningkatkan migrasi EPC pada darah tepi penderita penyakit jantung koroner stabil. Efek tertinggi tampak pada kelompok atorvastatin, diikuti kelompok simvastatin, dan rosuvastatin.   Kata kunci: Migrasi EPC, simvastatin, atorvastatin, rosuvastatin     Clinical Research   ABSTRACT   Effect of Statins on Endothelial Progenitor Cell (EPC) Migration from Peripheral Blood of Stable Coronary Artery Disease Patient   Tyagita Verdena Rani Savitri Yudi Her Oktaviono Djoko Soemantri     Background: Endothelial progenitor cell (EPC) participates in endothelial repair and new blood vessel growth. Cardiovascular pharmacotherapy has been shown to improve the amount and function of EPC in patients with cardiovascular risk. Many studies report that statins have a beneficial effect on EPC by increasing the number and function of EPC, including the migration function. Therefore, we conducted a study to analyze the effects of three different statins on EPC migration. Objective: To analyze the effect of statins on EPC migration from peripheral blood of stable coronary artery disease (SCAD) patient. Methods: This was an in vitro true experimental post-test only control group design. The MNCs were isolated from peripheral blood of SCAD patient and were cultured in Stemline II Hematopoietic Stemcell Expansion Medium in 3 days. Then samples were put into four groups, simvastatin 0.5 µmol/L, atorvastatin 0.5 µmol/L, rosuvastatin 0.5 µmol/L and control, then incubated for 48 hours. Immunocytochemical examination was performed to evaluate expression of CD34+. On the 5th day of culture, 5x105 cells per group were transferred to the upper chamber of the transwell system, then incubated for 1 day. Cells that migrated to the lower chamber of transwell system were calculated by automatic cell counter with typhan blue coloring. Data were analyzed by independent T-test and ANOVA. Results: The results of independent T-tests showed a significant increase in EPC migration in the simvastatin, atorvastatin, and rosuvastatin groups compared with the control group (234000 ± 1290.994, 265000 ± 1290.994, 203000 ± 1290. 994 vs 174071.43 ± 1426 785, p <0.05). EPC migration also differed between statin groups, where the highest effect was found in the atorvastatin group. EPC migration in the atorvastatin group was higher than the simvastatin group (265,000 ± 1290,994 vs 234,000 ± 1290,994, p <0.05), and simvastatin was higher than the rosuvastatin (234,000 ± 1290,994 vs 203000 ± 1290. 994, p < 0.05). Immunocytochemical examination showed a positive expression on CD34+. Conclusion: Statins increase EPC migration from peripheral blood of SCAD patient. Atorvastatin showed the highest EPC migration, followed by simvastatin, and rosuvastatin. Keywords: EPC migration, simvastatin, atorvastatin, rosuvastatin