The Effect Of Tgf-β1on Expression Chemokine And Its Receptor In Human Decidual Stromal Cells

In order to study the relationship between the transforming growth factor β (TGF-β) andchemokine which secreted by human stromal cells and to explore the mechanism of itsimmunomodulatory effects in human decidual stromal cells, In this experiment,we isolatedhuman trophoblast cells and human decidual stromal cells by the way of digestion with thetrypsin-collagenase-DNaseⅠ, the cells were cultured and identified in vitro. The expressionof chemokines and its receptors in early human decidual stromal cells after treatment withfour different concentrations of TGF-β1were detected by RT-PCR and Western-blot. Theresult showed that:1. Human trophoblast cells and human decidual stromal cells were isolated successfullyby the way of the digestion with trypsin, collagenase Ⅳa nd DNase Ⅰ. The cell culturesystem of two kinds of cells had been optimized, It is observing that human trophoblast cellswere a typical epithelial-like cell morphology, spreading growth by sheet, The staining ofcytokeratin was positive. Human decidual stromal cells were a fibroblast-like cellmorphology, the cell morphology gradually became blunt round after inducible with E2andP4.The cells outline were clear and the cytoplasmic particles were rich. The staining ofvimentin and prolactin were positive.2. The mRNA and protein level of chemokines(CXCL12, CXCL16and CX3CL1) weredecreased by RT-PCR and Western-blot after treatment with four different concentrations ofTGF-β1(0ng/mL,1ng/mL,5ng/mL and10ng/mL). On the contrary, the mRNA and proteinlevel of chemokine receptors were increased.At the mRNA level. Four concentrations ofTGF-β1can inhibit the expression of CX3CL1significantly(P＜0.01),10ng/mL TGF-β1caninhibit the expression of CXCL12significantly(P＜0.05),5ng/mL TGF-β1can inhibit theexpression of CXCL16significantly(P＜0.05),10ng/mL TGF-β1can promote the expressionof CXCL16significantly(P＜0.05). At the protein level,10ng/mL TGF-β1can inhibit theexpression of CX3CL1(P＜0.01) and CXCL16(P＜0.05) significantly, but can promote theexpression of CXCR6(P＜0.05).Conclusion: The method of separation,cultured and purification of human trophoblastcell and human decidual stromal cell laid the foundation for further study the molecular mechanism of decidualization in the study. TGF-β1can down-regulated the expression ofthe chemokine(CXCL12,CXCL16and CX3CL1) and up-regulated the expression of thereceptors(CXCR4and CXCR6). This study suggests that TGF-β1plays a significant role inregulating chemokine ligand/receptor expression and participate in immune tolerance ofmaternal-fetal interface by chemokines.