The Effect Of CD147Expressed In T Lymphocyte On Monocyte Chemotaxis And MMP-9Secreted By Monocyte In The Co-culture System

Background: Chronic inflammatory reaction can lead to the damage of arterial vesselwall and disturbance of endothelium function, both T lymphocytes and monocyteswere the significant inflammatory cells, which play an important role in the formationand development of the atherosclerosis. The highly expressed CD147T lymphocytewhether or not exist interaction with monocytes to influence the chemotaxis andsecretion MMPs in monocytes, and the CypA whether or not promoting chemotaxisand secretion MMPs in monocytes through CD147expressed in T lymphocyte, andthe cyclosporine A whether or not exist inhibitory action to this process, whichwaiting for further studying.Objective: Validate CD147expressed in T lymphocyte influence chemotaxis andsecretion MMPs of monocytes, to presume whether CypA could through CD147expressed in T lymphocyte influence chemotaxis and secretion MMPs of monocytes,and then reveal the possible immunologic mechanism in the formation anddevelopment of the atherosclerosis, providing new and effective target point toprevention and cure Atherosclerosis.Methods: The culture of Human T lymphocyte in vitro. specific siRNA targetingCD147gene was transfected into Jurkat cells which can inhibite CD147expressed inJurkat T lymphocytes, to validate the inhibited effect of CD147in proteins level andgene level after transfected siRNA. Constructed the co-culture system of humanJurkat T lymphocyte and THP-1monocyte in vitro, differently used CypA andcyclosporine A pretreated Jurkat T lymphocyte before co-culture, inhibited CD147express by siRNA, use CypA pretreated Jurkat T lymphocyte after siRNA specificinhibited CD147, CypA pretreated Jurkat T lymphocyte after cyclosporine A used;After every group of co-culture system differently co-culture48h to collect culture solution, which to detect by ELISA for MMP-9secreted by THP-1monocyte;chemotaxis experiment was tested use Millicell chemotaxis areole, to detect THP-1monocyte chemotaxis influenced by different intervention methods in everyco-culture system.Results: The siRNA to aim directly at CD147can specificly inhibite CD147expressed inJurkat Tlymphocyte,which compared with control group in siRNA interference groupthe mean fluorescence intensity of CD147in Jurkat cells was descended by（50.5±4.7）%（P<0.05）. The CD147mRNA and protein level were reduced by（38.57±1.55）%and （47.4±1.47）%respectively（P<0.05）, which consistentedwith the experiment requests. Compared with normal control group, the group ofJurkat inhibited CD147expressed by siRNA co-culture with THP-1, MMP-9secreted by THP-1monocyte and THP-1monocyte chemotaxis both decreasedobviously（p<0.05）; CypA pretreated Jurkat T lymphocyte co-culture with THP-1,MMP-9secreted by THP-1monocyte and THP-1monocyte chemotaxis bothincreased obviously（p<0.05）; but the group use cyclosporine A in Jurkat Tlymphocyte which co-culture with THP-1, MMP-9secreted by THP-1monocyte andTHP-1monocyte chemotaxis both decreased obviously（p<0.05）; the group of useCypA to stimulate Jurkat T lymphocyte which have been pretreated by cyclosporineA and the group of CypA pretreated Jurkat T lymphocyte after siRNA specificinhibited CD147then co-culture with THP-1monocyte, in these two groups, MMP-9secreted by THP-1monocyte and THP-1monocyte chemotaxis have no obviouslychanged（p>0.05）, the discrepancy was no statistical significance.Conclusions:1.Chemosynthesis siRNA could specificly inhibited expression of CD147gene inJurkat cells, providing groundwork for next research of CD147gene inAtherosclerosis.2.The co-culture system of human Jurkat T lymphocyte and THP-1monocyte couldconstruct successfully in vitro; The Jurkat T lymphocyte specific inhibited CD147bysiRNA could dncrease MMP-9secreted by THP-1monocyte and THP-1monocytechemotaxis in co-culture system, to illustrate Jurkat T lymphocyte expressed CD147 could influence monocyte chemotaxis and MMP-9secreted by monocyte in co-culturesystem.3. CypA could though CD147expressed in Jurkat T lymphocyte to influence THP-1monocyte chemotaxis and MMP-9secreted by THP-1in co-culture system, and whicheffect can be interrupted by cyclosporine A.