SUPLEMENTASI CALCITRIOL TERHADAP HOMEOSTASIS KALSIUM DAN AKTIFITAS OSTEOKLAS TULANG FEMUR TIKUS OVARIOHISTEREKTOMI

Hartiningsih, drh.,MP., Profesor Hastari Wuryastuty, M.Sc., Ph.D.

2012 | Disertasi | S3 Sain Veteriner

Penelitian ini dilakukan untuk mengkaji suplementasi calcitriol terhadap homeostasis Ca dan aktifitas osteoklas pada tikus ovariohisterektomi. Dua puluh tikus betina Wistar betina umur 8 minggu, dibagi 4 kelompok yaitu : tidak diovariohisterektomi tanpa suplementasi calcitriol (NT) dan tidak diovariohisterektomi diberi suplementasi calcitriol (ND), ovariohisterektomi tanpa suplementasi calcitriol (PT), dan ovariohisterektomi diberi suplementasi calcitriol (PD), masing-masing 5 tikus. Dua puluh sembilan minggu pasca operasi, setiap tikus dimasukkan kandang metabolik individu untuk studi balan. Selama studi balan (hari 5-11 studi balan), setiap pagi dilakukan pengumpulan feses, urin dan sisa pakan untuk pemeriksaan Ca. Pada hari ke-12 studi balan, tikus dietanasi, tulang femur kiri diambil untuk pemeriksaan immunohistokimia menggunakan monoklonal antibodi anti TRAP5b dan untuk pemeriksaan histopatologis. Hasil analisis menunjukkan bahwa tikus ND, PD, NT, dan PT berturut-turut mengkonsumsi Ca sebanyak (1142,82±89,32 mg/hari/kg), (1073,16±92,84 mg/hari/kg), (1054,73±49,91 mg/hari/kg), dan (882,62±118,61 mg/hari/kg). Absorpsi Ca melalui usus pada tikus ND, PD, NT dan PT berturut-turut (92,83±1,09%), (94,32±1,09%), (92,24±0,53%) dan (91,91±1,54%). Tikus ND, PD, NT dan PT mengekskresikan Ca dalam urin berturut-turut (2,12±0,34%), (2,31±0,82%), (3,8±2,45%) dan (1,51±0,43%). Ekskresi Ca dalam urin tikus PD tidak berbeda signifikan meskipun cenderung lebih tinggi, dan dari gambaran histopatologis, tikus PD mempunyai struktur ginjal normal. Retensi Ca pada tikus ND, PD, NT dan PT berturut-turut sebanyak (1037,33±72,73), (987,33±50,94), (932,05±24,43), dan (798,87±115,14). Dari gambaran histopatologis, ketebalan spikulum trabekula bagian metafisis tulang femur distalis berturut-turut paling tebal juga terlihat pada tikus ND, PD, NT dan PT. Dari gambaran immunohistokimia bagian metafisis tulang femur distalis terlihat positif TRAP5b berwarna coklat pada tulang trabekula antar rongga sumsum tulang dan pada permukaan spikulum trabekula pada tikus ND, PD, NT dan PT. Berdasarkan hasil penelitian disimpulkan bahwa suplementasi calcitriol 1) Meningkatkan 1,25- dihidroksivitamin D3, meningkatkan absorpsi Ca usus, namun ekskresi Ca dalam urin tidak berbeda, 2) Meningkatkan retensi Ca dan menurunkan aktifitas osteoklas, 3) Mencegah osteoporosis dan tidak menyebabkan kerusakan ginjal.

The obyective of the research was to study calcitriol supplementation on osteoporosis prevention of ovariohisterectomized Wistar rats by calcium homeostasisb and osteoclast activity. Twenty female of Wistar rats, 8 weeks of age were randomly divided into four groups (unovariohisterectomy without calcitriol supplementation (NT), unovariohisterectomy+ calcitriol supplementtation (ND), ovariohisterectomy without calcitriol supplementation (PT) and ovariohisterectomy+ calcitriol supplementation (PD) of five each. Twentynine weeks after the surgery, each rats was placed into individual metabolic cages for balance studies. At day 5 to 11 of the balance studies, the remaining food, urine, and feces were collected and recorded every day for Ca analyses. At day 12 of the balance studies, all rats were euthanationed, left femur was taken for histopatological examination using hematoxylin and eosin stain and for immunohistochemistry tartrate resistant acid phosphatase5b (TRAP5b) using monoclonal antibodi anti TRAP5b and was detected using streptavidin-biotin. The results showed that Ca consumption in rat groups ND, PD, NT, and PT were respectively (1142,82±89,32 mg/d/kg), (1073,16±92,84 mg/d/kg), (1054,73±49,91 mg/d/kg), and (882,62±118,61 mg/d/kg), and intestinal Ca absorption of rats in groups ND, PD, NT, and PT were respectively (92,83±1,09%), (94,32±1,09%), (92,24±0,53%) and (91,91±1,54%). The urinary Ca excretion of rats in groups ND, PD, NT and PT were respectively (2,12±0,34%), (2,31±0,82%), (3,8±2,45%.), and (1,51±0,43%). The urinary Ca excretion of rats in groups PD was not significantly different although they were tend to increased. Histopathological analysis of kidney of rats in groups PD showed normal structure. Calcium retention of rats in groups ND, PD, NT and PT were respectively (1037,33±72,73), (987,33±50,94), (932,05±24,43), and (798,87±115,14). Tartrate resistant acid phosphatase5b activity was decreasing respectively in rats of ND, PD, NT and PT. Trabecular speculum thickness was thicker respectively in rats of ND, PD, NT and PT. Base on the results of the present study, it was concluded that calcitriol supplementation effected : 1) In increased 1,25-dihidroksivitamin D3, increased intestinal calcium absorption and undifferent urinary Ca excretion, 2) In increased Ca retention and reduced osteoclast activity, and 3) In osteoporosis prevention and safe for the kidney.

Kata Kunci : Calcitriol, ovariohisterektomi, homeostasis Ca, osteoklas, osteoporosis