Experimental Study On The Effect Of Guilu ErXian Glue On Expression Of Type â…  Collagen Gene Of Osteoblast

Background:Studies have show that type I collagen is closley related to the osteoporosis. Type I collagen is genetic encoded by Collal and Colla2 gens.which can be regulated by transforming growth factor beta(TGF-β). TGF-β1 and TGF-β2 are two isomers of TGF-βs. Traditional Chinese medicine Guilu ErXian Glue can be very efficiency during the treatment of OP. Instead of study about the type I collagen, past research was mostly focused on the clinical observation.Purpose:To study the effect Guilu ErXian Glue has on the expression of type I collagen gene of osteoblast, and to explore the machanism of the treatment of OP using Guilu ErXian Glue.Methods:100 SD mouses were randomly divided into control group, Guilu ErXian Glue group(low dose), Guilu ErXian Glue group (medium dose), Guilu ErXian Glue group(high dose),calcitriol group and control group(without any treatments), lavage 1 time a day,7days in a row,2 hours after last delivery, take abdominal aortic blood and make them into drug-containing osteoblasts. Find the effect of these drug-containing serum on the proliferation and apoptosis of the osteogenesis cell using MTT method and the observation of cell morphology. And using Western Blot method to detect the express level of type I collagen and TGF-P protein, PCR to detect the mRNA expression of type I collagen and TGF-β.Results:(1)MTT results shows that:those Guilu ErXian Glue groups have no obvious proliferation of osteoblast. The calcitriol group has significant increment of osteoblasts(P<0.05). Living cell dyeing shows that those groups all have no significant apoptosis of osteoblasts.(2)The Western Blot results suggest:for 24 hours, high dose group of Guilu ErXian Glue can increase the opsteoblast synthesis COL1A1, COL1A2 protein(P<0.05). For 48 hours, calcitriol group, low, medium and high dose groups of Guilu ErXian Glue can increase the osteogenesis COL1A1 and COL1A2 protein expression(P<0.01), medium and high dose group of COL1A1 and COL1A2 protein expression approach. For 24 hours and 48 hours, calcitriol group of TGF-β1, TGF-β2 protein expression without significantly increased; P>0.05. FOR 24 hours, the high dose group of Guilu ErXian Glue about osteoblasts TGF-β1, TGF-β2 expression was significantly role in promoting; For 48 hours, low, medium and high dose group of Guilu ErXian Glue can make the osteoblast TGF-β1, TGF-β2 protein expression increased, medium and high dose group of Guilu ErXian Glue has no statistically significant difference (P>0.05). For 48 hours, low, medium and high dose groups of Guilu ErXian Glue can increase osteoblast protein expression. (p<0.01) There was no statistically significant difference between medium and high dose group(P>0.05).(3)RT-PCR method showed:calcitriol and the groups of Guilu ErXian Glue can make the osteoblast COL1A1 and COL1A2 mRNA level increased (P<0.01);The groups of Guilu ErXian Glue COL1A1 rubber group, COL1A2mRNA level increased obviously(P<0.05); Guilu ErXian Glue high dose group is set of COL1A1 and COL1A2mRNA levels has no obvious increase than the medium group. The TGF-β1 and TGF-β2 mRNA level of calcitriol group has no significant change; the TGF-β1 and TGF-β2 mRNA level of All The Guilu ErXian Glue groups were increased,and the two group of medium and high dose group has the same amount of increment(P>0.05).Conclusion:Guilu ErXian Glue can promote the expression of type Ⅰ collagen gene,which may be related to the regulation TGF-β factors.