Expression of PAPP-A, IGF-I and Their Effects on the Cytological Functions of siRNA Lentiviral Vector of hCASMCs IGF-I after Inflammatory Factor

LIU Hong-sheng, ZHAO Xiao-dong, SU Qin. et al

Abstract

To evaluate expression of PAPP-A, IGF-I and the effect of TNF-α, IL-1β on the cytological functions of hCASMCs with IGF-I gene silencing after inflammatory factor, in order to further study on the action of IGF axis hormone in the rupture of astable atheroxclerosis plaque. Methods A RNA interference (RNAi） aimed at the gene of IGF-I was carried out to have an eukaryon transfection to hCASMCs. When the IGF-I-shRNA-hCASMC were treated by TNF-α, IL-1β, IGFBP4, the expression of PAPP-A and IGF-I were detected with Western blot and ELISA. And then, the effect of TNF-α,IL-1β,IGFBP4 on the proliferation of IGF-I-shRNA-hCASMC were assessed by MTT assay and changing in cell cycle and apoptosis were evaluated by using flow cytometry. Results Significant positive expression of PAPP-A in hCASMCs which were treated by TNF-α+IL-1β or TNF-α+IL-1β+IGFBP4 in Blank control (CON), Negative control (NC) and RNAi group were observed, but lower expression in the RNAi group than that in CON and NC groups. However, there was no positive expression of PAPP-A in hCASMCs of CON, NC, RNAi group treating without TNF-α+IL-1β or TNF-α+IL-1β+IGFBP4. The expression of IGF-I in hCASMCs of CON, NC, RNAi group treated with TNF-α+IL-1β+IGFBP4 were greater than that only with or without TNF-α+IL-1β treatment. In RNAi group, the A570 decreased when the hCASMCs treated with TNF-α+IL-1β+IGFBP4 and significant lower than that in hCASMCs treated with TNF-α+IL-1β. When the hCASMCs were treated with TNF-α+IL-1β or TNF-α+IL-1β+IGFBP4, the rate of apoptosis significantly increased in RNAi group, which was significantly higher than that in CON group and NC group. In addition, in RNAi group, the rate of apoptosis in hCASMCs treated with TNF-α+IL-1β+IGFBP4 was significant higher than that in hCASMCs treated only with TNF-α+IL-1β. Conclusion When the IGF-I-shRNA-hCASMCs were stimulated by some inflammation factors, its proliferation decreased but the apoptosis enhanced. So the activated IGF-I level in local microebvironnment increased, which may cause the descend of cell proliferation and the increasing of apoptosis.

 

Keywords: Inflammation factors, Human coronary artery vascular smooth muscle cells, RNA interference, IGF axis, Cytological functions 

 

Full Text:

PDF


References


Bayes-Genis А, Conover С A. Overgaard MT. et al. Pregnancy-associated plasma protein A as a marker of acute coronary syndromes. N Engle J Med. 2001; 345 ( 14 ): 1022- 1029.

Bayes-Genis A, Conover CA. Schwartz RS. The insulin-like growth factor axi:a review of atherosclerosis and restenosis. Circ Res.2000;86(2): 125-130.

Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 7 ( 72) : 248-254.

Gaidamauskas K. Gyrup C. Boldt HB,et al. IGF dependent modulation of IGF binding protein (IGFBP) proteolysis by pregnancy-associated plasma protein-A ( PAPP-A): multiple PAPP-A-IGFBP interaction sites. Biochim Biophys Acta.2013; 1830(3) :2701-2709.

Woelfle J, Roth CI-. Wunsch R. et al. Pregnancy-associated plasma protein A in obese children;relationship to markers and risk factors of atherosclerosis and members of the IGF system. Kur J Endocrinol.2011; 165(4) :613-622.

Harrington SC'. Simari RD. Conover CA. Genetic deletion of pregnancy-associated plasma protein A is associated with resistance to atherosclerotic lesion development in apolipoprotein E-deficient mice challenged with a high-fat diet. Circ Res.2007; 100(12): 1696-1702.

Conover CA. Mason MA. Bale LK. et al. Transgenic overexpression of pregnancy-associated plasma protein-A in murine arterial smooth muscle accelerates atherosclerotic lesion development. Am J Physiol Heart Circ Physiol. 2010; 299( 2): H284-91.

Conover CA. Harrington SC. Hale LK. Differential regulation of pregnancy associated plasma protein-A in human coronary artery endothelial cells and smooth muscle cells. Growth Horm IGF Res.2008; 18(3):213-220.

Conover CA. Bale LK. Harrington SC', et al. Cytokine stimulation of pregnancy-associated plasma protein A expression in human coronary artery smooth muscle cells: inhibition by resveratrol. Am J Physiol Cell Physiol.2006;290 (10):Cl83-088.

Higashi Y. Sukhanov S. Anwar A. et al. Aging, atherosclerosis, and IGF-1. J Gerontol A Biol Sci Med Sci. 2012;67(6):626-639.

Harrington SC. Genetic deletion of pregnancy -associated plasma protein A is associated with resistance to atherosclerotic lesion development in apolipoprotein E-deficient mice challenged with a high-fat diet. Circ Res.2007; 100( 12) : 1696- 1702.

Mason KJ. Grell JA.Wan J. et al. Insulin-like growth factor (IGF)- 1 and IGF- II contribute differentially to the phenotype of pregnancy associated plasma protein-A knock-out mice. Growth Horm IGF Res,2011;21(5):243-247.

Shai SY. Sukhanov S. Higashi Y. et al. Smooth muscle cell-specific insulin-like growth factor-1 overexpression in Apoe— /— mice does not alteratherosclerotic plaque burden but increases features of plaque stability. Arterioscler Thromb Vase Biol.2010;30( 10): 1916-1924.

Tong X. Zitserman D, Serebriiskii I. et al. Numb independently antagonizes Sanpodo membrane targeting and Notch signaling in Drosophila sensory organ precursor cells. Mol Biol Cell.2010;21 (5):802-810.

Rosi MC. Luccarini I. Grossi C. et al. Increased Dickkopf-1 expression in transgenic mouse models of neurodegenerative disease. J Neurochem.2010; 112(6): 1539-1551.

Angulo K.Casado V.Mallol J, et al. Al adenosine receptors accumulate in neurodegenerative structures in Alzheimer disease and mediate both amyloid precursor protein processing and tau phosphorylation and translocation. Brain Pathol.2003; 13(4):440-451.

Kumar A A. Kumar SR. Narayanan R. et al. Autologous bone marrow derived mononuclear cell therapy for spinal cord injury: a phase I / II clinical safety and primary efficacy data. Exp Clin Transplant, 2009; 7(4): 241-248.

Sun J. Gao Y. Yang L. et al. Neural-tube-derived neuroepithelial stem cells: a new transplant resource for Parkinson's disease. Neuroreport,2007; 18(6):543-547.

Avraham Y. Dayan M. I-assri V, et al. Delayed leptin administration after stroke induces neurogenesis and angiogenesis. J Neurosci Res.2013;91(2); 187-195.

Guo Z. Jiang H. Xu X. et al. Lept in-mediated cell survival signaling in hippocampal neurons mediated by JAK STAT3 and mitochondrial stabilization. J Biol Chem. 2008; 283 (3): 1754- 1763.

Petersen PH. Zou K. Krauss S. et al. Continuing role for mouse Numb and Numbl in maintaining progenitor cells during cortical neurogenesis. Nat Neurosci.2004;7(8):803-811.


Refbacks

  • There are currently no refbacks.