Effects Of Alzet Mini-osmotic Pumps Controlled Release Of IGFBPrP1 On Mice Hepatic And Lung Tissues

BackgroundInsulin-like growth-factor binding protein-related protein 1 is a soluble cell adhesion glycoprotein, Secreted by the tumor cells, vascular endothelial cells, smooth muscle cells and fibroblasts and other cells. IGFBPrPl protein was widely expressed in various human cells tissues and plays an important role from Lopez-Bermejo and Ma's study, it can regulate epithelial cell and fibroblast cell growth, proliferation, adhesion, and promote angiogenesis and stimulate endothelial cells to produce prostacyclin, induce apoptosis etc. Protein function on IGFBPrP1 is not completely clear currently. Lixin Liu discovered that HSC could be activated by IGFBPrP1 in vitro. Anti-IGFBPrP1 antibody can suppresses the progression of hepatic fibrosis through many ways, including inhibiting the activation of HSC and reducing hepatocyte apoptosis and so on. IGFBPrP1 was a new pathogenic factor in the formation of liver fibrosis.IGFBPrP1 biological function needs to be further studied:On the one hand, IGFBPrP1 injected intraperitoneally administered short half-life, the peak drug concentration, fast metabolism, the function of proteins IGFBPrP1 can not really be understood; On the other hand, IGFBPrP1 could cause liver fibrosis, whether it affects other organs and the possible mechanism has yet not be clear. To clarify this problem, we design this issue.ObjectiveIGFBPrP1 was released at a continuous constant dose by Alzet mini-osmotic pump which was implanted subcutaneously of wild-type mice, Compare the effects of IGFBPrP1 on mice liver and lung tissues and explore the mechanism.MethodsTwenty-four male C57BL/6 wild-type mice were randomly divided into normal group, sham-operated group and Operation group. We used rmIGFBPrP1 which was released by capsule osmotic pump to make the model. Sham-operated group was only treated by control operation. HE and Picrosirius red staining were used to evaluate pathomorphological changes and collagen generation. The expression and distribution of IGFBPrP1,Collagen-III Smad3,P-Smad2/3 protein in mice hepatic and lung tissues were tested by immunohistochemistry methods. Apoptosis of hepatic cells were investigated by TUNEL.Results (1) The picric acid-Sirius red staining analysis:collagen content in liver tissue of operation group significantly increased than normal group and sham group (0.46%±0.09%vs 1.27.%±0.10%,0.45%±0.08%vs 1.27%±0.10%, F=597.797, P<0.01), collagen fiber content in lung tissue of operation group no significant difference than the other two groups (0.35%±0.09% vs 0.36%±0.12%,0.37%±0.12% vs 0.36%±0.12%, F=0.344, P>0.05)。(2) The IHC staining analysis: The expression of IGFBPrP1,Collagen III and p-Smad2/3 except Smad3 in liver tissue of operation group significantly increased than normal group and sham group. (IGFBPrP1:0.26±0.04 vs 1.77±0.64,0.25±0.04 vs 1.77±0.64, f=44.064, P<0.01; Collagen III:0.45±0.04 vs 2.59±0.29,0.43±0.05 vs 2.59±0.29, F=431.641, P<0.01; Smad3:0.29±0.06 vs 1.20±0.27,0.34±0.04 vs 1.20±0.27, F=292.114, P<0.01; p-Smad2/3:0.53±0.07 vs 2.44±0.32,0.51±0.08 vs 2.44±0.32, F=114.251, P<0.01)。The expression of IGFBPrP1,Collagen III,Smad3 and p-Smad2/3 in lung tissue of operation group no significant difference than the other two groups. (IGFBPrP1:0.32±0.05 vs 0.30±0.07,0.29±0.06 vs 0.30±0.07, F=0.505, P>0.05; Collagen III:0.93±0.07 vs 0.89±0.02,0.85±0.01 vs 0.89±0.02, F=0.973, P>0.05; Smad3:0.26±0.03 vs 0.25±0.04,0.24±0.09 vs 0.25±0.04, F=0.150, P>0.05; p-Smad2/3: 0.25±0.06vs0.26±0.05,0.28±0.06 vs 0.26±0.05, F=0.255, P>0.05)。(3) The TUNEL staining analysis:More TUNEL positive cells were found in operative group than normal group and sham-operated group (6.21%±2.83%vs 34.14%±11.04%, 7.13%±2.59%vs 34.14%±11.04%, F=114.251, P< 0.01; 6.10%±2.23%vs 32.23%±10.60%,5.71%±2.87%vs32.23%±10.60%, F=114.251,P<0.01)。Conclusion1. Exogenous IGFBPrP1 released at a continuous constant dose may induce hepatic fibrosis of mice. On the same condition, the effect of IGFBPrP1 on mice lung tissue was weaker than that on hepatic tissue and pulmonary fibrosis was not found.2. IGFBPrP1 has a selectively functions on mice tissue.