Observation Of Influence That The Xiehuo Pingtu Powder Has On Retrobulbar Fibroblasts After Stimulated By IFN-γ

Objective:(1)Observe the influence of Xiehuo Pingtu Powder-containing serum has on the propagation of retroocular fibroblast,the synthesis of HA and type(Ⅰ) collagen and type(Ⅲ) collagen,the activity of MMP-1、MMP-2、TIMP-1、TIMP-2,and the expression of ICAM-1、HLA-DR、CD40;(2)Discuss the mechanism of Xiehuo Pingtu Powder treating TAO.Method: Retroocular fibroblast taken from normal subjects was primary cultured and subcultured in vitro,then 2-8 generation cellule was authenticated and sectionalizated to be experimented after confirming as retroocular fibroblast.The control group was cultured in normal medium and the blank serum group was cultured in medium containing 20% serum of normal rats,while the stimulating group was cultured in medium containing 200u/ml IFN-γ.The Xiehuo Pingtu Powder-treated group was cultured in medium containing 200u/ml IFN-γ and 20% Xiehuo Pingtu Powder-medicined serum.Then measured the proliferation of RFs, synthesis of HA and type(Ⅰ) collagen and type(Ⅲ) collagen,the activity of MMP-1、MMP-2、TIMP-1、TIMP-2,and the expression of ICAM-1、HLA-DR、CD40 in these groups with MTT、ELISA、Immunohistochemistry、Flow Cytometry respectively.Then all the datas was statistically analysised.Results:(1)The values of OD were mesured with MTT and to evaluate the proliferation of RFs in all groups.There is no statistical difference between the control group and the blank serum group(P>0.05),and the OD values of the stimulating group was bigger than that of the control group and the blank serum group and the there is some statistical significance(P<0.05).The OD values of Xiehuo Pingtu Powder-treated group significantly decreased than that of the control group and there is some statistical significance(P<0.05).(2)The synthesis of HA and type(Ⅰ) collagen and type(Ⅲ) collagen was detected with ELISA.The synthesis of HA and type(Ⅰ) collagen and type(Ⅲ) collagen of the stimulating group was more than that of the control group and the blank serum group and there is some statistical significance(P<0.05).The synthesis of the Xiehuo Pingtu Powder-treated group was less than that of the control group and there is some statistical significance(P<0.05).(3)The activity of MMP-1、MMP-2、TIMP-1、TIMP-2 was mesured with Immunohistochemistry.There is no statistical significance between the control group and the blank serum group(P>0.05).The activity of MMP-1、MMP-2、TIMP-1、TIMP-2 of the stimulating group was lower obviously than that of the control group and the blank serum group and there is some statistical significance(P<0.05).The activity of MMP-1、MMP-2、TIMP-1、TIMP-2 of the Xiehuo Pingtu Powder-treated group was higher than that of the stimulating group and there is some statistical significance(P<0.05).(4)The activity of MMP-1、MMP-2、TIMP-1、TIMP-2 was mesured with Immunohistochemistry.There is no statistical significance between the control group and the blank serum group(P>0.05).The activity of MMP-1、MMP-2、TIMP-1、TIMP-2 of the stimulating group was lower obviously than that of the control group and the blank serum group and there is some statistical significance(P<0.05).The activity of MMP-1、MMP-2、TIMP-1、TIMP-2 of the Xiehuo Pingtu Powder-treated group was higher than that of the stimulating group and there is some statistical significance(P<0.05).(5)The expression of ICAM-1、HLA-DR、CD40 was merured with Flow Cytometry.There is no statistical significance between the control group and the blank serum group(P<0.05).The expression of the stimulating group is more than that of the control group and the blank serum group and there is some statistical significance(P<0.05).The expression of the Xiehuo Pingtu Powder-treated group was less than that of the stimulating groupand there is some statistical significance(P<0.05).Conclusion: Xiehuo Pingtu Powder treats TAO by inhibiting the propagation of retroocular fibroblast and The expression of ICAM-1、HLA-DR、CD40 to regulating the balance of MMPs/TIMPs and inhibiting the synthesis of HA and type(Ⅰ) collagen and type(Ⅲ) collagen.