VITAMIN D AKTIF (1,25(OH)2D3) MENCEGAH FIBROSIS GINJAL PADA MENCIT SWISS WEBSTER JANTAN DENGAN UNILATERAL URETERAL OBSTRUCTION (UUO) Kajian terhadap Kadar Transforming Growth Factor – Beta1 (TGF-β1), Fraksi Area Kolagen dan Fraksi Area Miofibroblast di Ginjal

KHUSNUL MUFLIKHAH, Prof.dr. Sri Kadarsih Soedjono, M.Sc., Ph.D., AIFM; dr. Nur Arfian, Ph.D

2015 | Tesis | S2 Ilmu Kedokteran Dasar dan Biomedis

Latar Belakang: Fibrosis tubulointerstisial ginjal merupakan fase akhir penyakit ginjal yang dapat mengakibatkan End Stage Renal Disease (ESRD) dan mengganggu homeostasis tubuh. TGF-β1 telah diketahui memiliki peran penting dalam proses fibrogenesis ginjal, melalui jalur yang melibatkan protein Smad. Unilateral Ureteral Obstruction (UUO) merupakan model yang representatif untuk fibrosis tubulointerstisial ginjal. Vitamin D diketahui memiliki efek renoprotektif. Mekanisme vitamin D dalam mencegah fibrosis diantaranya melalui penghambatan terhadap TGF-β1, dan pencegahan aktivasi miofibroblast. Metode: Penelitian ini merupakan penelitian kuasi eksperimental dengan rancangan post test only with control group dengan subjek 25 ekor mencit Swiss webster jantan dewasa yang dibagi dalam 5 kelompok. Setelah pemberian vitamin D selama 14 hari, diperiksa kadar TGF-β1 ginjal dengan metode solid phase sandwich ELISA, fraksi area kolagen dengan pewarnaan picrosirius red, dan fraksi area miofibroblast dengan pewarnaan imunohistokimiawi menggunakan antibodi α-sma. Penghitungan fraksi area dilakukan dengan software ImageJ Hasil: Rerata±SEM fraksi area kolagen kelompok SO (4,64±0,49)%, UUO (19,97±1,82)%, UD1 (10,78±3,95)%, UD2 (10,6±1,14) %, dan UD3 (8,71±0,62)%, dengan nilai p<0,001. Median (min-maks) fraksi area miofibroblast kelompok SO 0,66(0,29-7,49)%, UUO 21,17(17,30-25,99)%, UD1 7,09(4,57-10,23)%, UD2 5,28(2,66-11,37) %, dan UD3 5,10(2,14-7,46)%, dengan nilai p=0,004. Median (min-maks) kadar TGF-β1 kelompok SO 5,72(4,93-6,93)pg/mL, UUO 76,07(46,71-113,23) pg/mL, UD1 5,39 (4,93-6,05) pg/mL, UD2 4,93(4,20-6,05) pg/mL, dan UD3 4,77(3,92-9,07) pg/mL, dengan nilai p=0,009. Terdapat korelasi positif (p<0,05) antara kadar TGF-β1 dengan miofibroblast, fraksi area kolagen dengan fraksi area miofibroblast, masing-masing dengan kekuatan korelasi sedang. Kesimpulan: Pemberian Vitamin D mencegah fibrosis ginjal pada mencit jantan dengan UUO

Background: Renal tubulointerstitial fibrosis is the final phase of kidney disease which can cause End Stage Renal Disease (ESRD) and disturb the body homeostasis. TGF-1 is known as its important role in renal fibrogenesis process through protein Smad-involved pathway. Unilateral Ureteral Obstruction (UUO) is a representative model for renal tubulointerstitial fibrosis. Vitamin D is known to have the renoprotective effect. The mechanisms of vitamin D in preventing fibrosis are through resistance towards TGF-1 and prevention of myofibroblast activation. Method: This is a quasi experimental research with post test only with control group design. Subjects of male adult Swiss Webster mice (n = 25) were divided into 5 groups. After vitamin D application for 14 days, kidney TGF-1 level was tested by using solid phase sandwich ELISA method, area fraction of collagen by using picrosirius red staining, and area fraction of myofibroblast by using antibody -SMA of immunohistochemical coloration. Enumeration of area fraction was done by using software Image J. Results: Mean±SEM of TGF-1 level groups of SO (6,01±0,37) pg/ ml, UUO (77,92±12,91) pg/ ml, UD1 (5,44±0,23) pg/ ml, UD2 (5,07±0,31) pg/ ml, and UD3 (5,66±0,93) pg/ ml, with p value =0,009. Mean±SEM of area fraction of collagengroups of SO (4,64±0,49)%, UUO (19,97±1,82)%, UD1 (10,78±3,95)%, UD2 (10,6±1,14) %, and UD3 (8,71±0,62)%, with p value <0,001.Mean±SEM ofarea fraction of myofibroblast groups of SO (2,00±1,38%)%, UUO (21,51±1,49)%, UD1 (7,13±0,97)%, UD2 (6,08±1,56)%, and UD3 (5,16±0,87)%, with p value <0,001. There were positive correlation (p< 0,05) between TGF-1 level with myofibroblast, and area fraction of collagen with area fraction of myofibroblast, each with medium correlation strength. Conclusion: The application of vitamin D can prevent renal fibrosis in male mice by UUO.

Kata Kunci : vitamin D, fibrosis tubulointerstisial ginjal, TGF-β1, kolagen, miofibroblast