Bisphosphonate가 조골세포 분화에 미치는 영향
EFFECT OF BISPHOSPHONATE ON OSTEOBLAST DIFFERENTIATION
소속 상세정보
이인순/In Soon Lee
김현정/김영진/남순현/류현모/Hyun Jung Kim/Young Jin Kim/Soon Hyeun Nam/Hyun Mo Ryoo
KMID : 0358920000270020309
Abstract
kabstract :
본 실험은 bisphosphonate가 조골세포 분화 및 파골세포 분화에 미치는 영향을 알아보고
자, etidronate와 alendronate를 조골세포에 투여하여 조골세포 전사인자인 Cbfal, 조골세포
표시인자의 발현, 석회화된 골결절 형성을 평가하였다.
Bisphosphonate가 조골세포의 석회화된 골결절 형성에 미치는 영향을 평가하기 위하여
배양액에 10-6, 10-5, 10-4 M의 etidronate 및
10-8, 10-7, 10-6M의 alendronate를 첨가하였으
며, 배양 15일 후에 alizarin red로 염색하여 관찰하였다. 또 조골세포의 분화에 미치는
bisphosphonate의 영향을 평가하고자 백서 두 개관에서 얻은 조골세포에 etidronate
10-6, 10-5, 10-4M 및 alendronate
10-6M을 투여하고 배양 8일 후 총RNA를 수집하였고, 전기영동 및 Northern
blot hybfidization하여 Cbfal, alkaline phosphatase, type I collagen, osteopontin, osteocalcin
의 발현을 조사하였다.
이상의 실험결과 다음과 같은 결론을 얻었다.
1. Etidronate는 농도 의존적으로 골결절 석회화를 억제하였으나, alendronate는 골석회화
를 억제하지 않았다.
2. Etidronate는 Cbfal의 발현은 농도 의존적으로 억제하였으나, alendronate는 오히려 촉
진하였다.
3. Etidronate는 type I collagen, osteocalcin 및 osteopontin의 발현을 농도 의존적으로 억
제하였으나 alendronate는 오히려 증가시켰다.
4. Alkaline phosphatase의 발현은 사용된 etidronate와 alendronate에 의해 영향 받지 않
았다.
이상의 결과에서 etidronate는 조골세포의 전사인자인 Cbfal의 발현을 억제하며, 이에 의
하여 조골세포의 분화표지인자인 type I collagen, osteopontin 및 osteocalcin의 합성이 억제
되고, 결과적으로 석회화된 골결절의 형성을 억제하는 것으로 사료된다.
#초록#
-Abstract-
Bisphosphonates inhibit bone resorption in vivo and in vitro. Currently proposed
mechanism of action of bisphosphonates involves both direct effect on osteoclasts and
indirect effect through the mediation of osteoblasts. Recent understanding of molecular
mechanism of osteoclastogenesis indicates that osteoclast differentiation is quite tightly
regulated by signaling molecules from differentiating osteoblasts. Therefore this
investigation was designed to elucidate the effect of bisphosphonate on osteoblast
differentation. For this purpose, in vitro effects of etidronate and alendronate on the
expresstion of Cbfal, a master control gene of osteoblast differentiation, several bone
marker genes, and formation of calcified nodules were evaluated.
To evaluate the effect of bisphosphonate on calcified nodule formation, osteoblasts
isolated from rat calvaria were cultured in a-MEM containing 10-4,
10-5, 10-6M of etidronate or 10-6,
10-7, 10-8M of alendronate for 15 days, and then stained by
alizarin red to determine mineralization. To evaluate the effect of bisphosphonate on
osteoblast differentiation, osteoblast cells were cultured in a-MEM containing
10-4, 10-5, 10-6M of etidronate or
10-6M of alendronate for 8 days. And then total RNA was extracted and
northern blot analysis was done to examine the expression of Cbfal, type I collagen,
alkaline phosphatase, osteopontin and osteocalcin.
The results were as follows :
1. Etidronate suppressed the calcification of bone nodule in dose dependent manner,
while alendroate didn't.
2. The expression of Cbfal was decreased dose dependently by etidronate, but
increased by alendronate.
3. Etidronate suppressed the expression of type I collagen, osteopontin and osteocalcin
in dose dependent manner, however alendronate promote the expression of osteoblast
marker gene.
4. The expression of alkaline phosphatase was not affected either etidronate nor
alendronate
These results suggest that etidronate suppressed the expression of Cbfa 1 in dose
dependent manner, and consequently the expression of osteoblast marker genes, such as
type I collagen, osteopontin and osteopontin and osteocalcin were also suppressed in
similar manner. And finally this decreased expression of osteoblastic marker gene
prevent calcified bone nodule formation.
키워드
bisphosphonate; 조골세포; Cbfal; Type I 교원질; alkaline phosphatase; osteopontin; osteocalcin; 석회화; Bisphosphonate; osteoblast; Cbfal; Type I collagen; Alkaline phosphatase; Osteopontin; Osteoclacin; Calcified bone nodule;
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