Effect And Mechanism Of Platelets On The Enhancing Inflammatory Response In Trauma Patients By Infusing Banked Red Blood Cell

Background: Allogeneic transfusion (allogeneic blood transfusion, ABT) inpatients with clinical rescue process play an important role, but allogenic bloodtransfusion in life-saving at the same time, will be complicated by immune-relatedside effects. Transfusion-related immunomodulation (transfusion-relatedimmunomodulation, TRIM) is the input of allogenic blood cause immune systemchanges to immune suppression syndrome characterized by common clinicalconcurrency TRIM led to the serious consequences of immunosuppression forpostoperative infection and tumor recurrence and metastasis. Suspended red bloodcells in the preservation process, contained in white blood cells and platelets canrelease a variety of biologically active substances, the concentration of thesebiologically active substances with the extension of the retention time increased, thesebiologically active substances into the body may cause the recipient immune status ischanged, leading to the consequences of TRIM.TRIM occurrence of the exact mechanism is unclear. The current study is known,a variety of stimulatory signals must be combined with their corresponding target cellmembrane receptors, and through its specific signal transduction pathways play abiological role. Recent studies have found that platelets play an important role notonly in the process of hemostasis and coagulation;platelets also participate in a lot ofinflammation. Platelets are the smallest blood cells in the peripheral blood, and partoff the formation of megakaryocyte cytoplasm containing dense granules and αparticles, if activated, can release a variety of cytokines and growth factors, such assoluble CD40ligand (of sCD40L), tumor necrosis factor α (TNF-alpha), interleukin-1β (IL-1β), transforming growth factor beta (of TGFβ), RANTES et al. In the redblood cell preservation process, platelets are very easily activated, thus releasing alarge number of bioactive factors, these biologically active factors in the TRIM, fewreports of the relevant literature.Objective: To explore the role and mechanism of the inflammatory response inpatients with platelet transfusion inventory erythrocytes enhanced.Method 1. An inventory red blood cell supernatant preparation:Take three fresh collection of red cells suspension bags joints mixed with sterilebonding machine with sterile empty bags placed in the refrigerator at4℃preservation.7d,21d,35d bonding machine with sterile and sterile empty bags jointspecimens from sample to squeeze into a sterile empty bag,4000g centrifugation, redblood cell upper liquid obtained contains a small amount of red blood cell supernatant,about130ml. This supernatant was poured into sterile EP tube, to10000gcentrifugation to remove red blood cells, red blood cell supernatant of sterile manner.This supernatant was distributed into six points, placed in-80℃refrigerator..2. Experimental groups:Take150ml platelets of the new collection with the same type of plasma dilution,sterile bonding machine with platelet storage bags connected to squeeze into the40mlplatelets, and so repeat12times. Choose one of four bags of platelets sterile mannerall join7d inventory erythrocyte supernatant5ml, then three bags of sterile mannerconcentration of10ng/ml,100ng/ml to1000ng/ml LPS and mark45ul better.Sterile manner all add28d of inventory re-election four bags of platelets in theremaining platelets erythrocyte supernatant5ml, then three bags of sterile manner byadding45ul of the concentration of10ng/ml,100ng/ml to1000ng/ml LPS andlabeled.4bags of platelet re-election in the remaining platelets sterile manner alljoined the35d stock erythrocyte supernatant5ml, then three bags of sterile manner byadding45μl concentrations of10ng/ml and100ng/ml,1000ng/ml LPS and marked.Finally, the rest of anything in a bag of platelets, namely:Group A: control group, diluted apheresis platelet40ml.Group B: Add5ml7d or21d or35d, red blood cell supernatant45ml in platelets.Group C: by adding5ml7d or21d or35d red blood cell supernatant and45μlconcentration of10ng/ml of LPS in apheresis platelet45ml.Group D: Add5ml7d or21d or35d red blood cell supernatant and45μlconcentration of100ng/ml LPS machine platelets45ml.Group E: to join5ml7d or21d or35d, red blood cell supernatant and45μlconcentrations of LPS machine1000ng/ml platelets45ml.3. The results of detection:At3h,6h,24h sterile manner in the platelets remove10ml respectively, of which1ml for platelet apoptosis, activation detection,2ml for cytokine detection and the restfor platelet aggregation and hypotonic shock detection. CD62P and platelet apoptosis using flow cytometry; platelet hypotonic shock response using spectrophotometricdetection; platelet aggregation rate of platelet aggregation was detected; ofCD40L,5-HT, of TGF-β1, RANTES, IL-1β and TNF-α using the enzyme-linkedimmunosorbent assay.4. Results1. The results of sCD40LAt the same point in time, concentration of sCD40L with the stock red blood cellsupernatant time increased (P <0.05); in the same stock erythrocyte supernatantconcentration of sCD40L with platelet save time increased (P <0.05); under the samecondition sconcentration of sCD40L increased with increase in concentration of LPSsolution (P <0.05).2. The results of5-HTAt the same time point,5-HT concentration with the stocks in the same timeextend and increase red blood cell supernatant time (P <0.05); in the same stockerythrocyte supernatant,5-HT concentration increased (P <0.05) with the plateletstorage time extended;,5-HT concentrations under the same conditions with the LPSconcentration in the solution increases (P <0.05).3. The results of TGF-β1At the same time point, TGF-β1concentration with the stocks, red blood cellsupernatant time to extend the increased (P <0.05) in the same stocks in the red bloodcell supernatants, TGF-β1release in3h,6h at no difference (P>0.05).7d red bloodcell supernatant stimulated the release of TGF-β1and with time (P>0.05),21d,35dred blood cell supernatants to stimulate the release TGF-β1in24h at significantlyincreased (P <0.05); the same time point, TGF-β1concentration with the increase ofLPS concentration in solution increased (P <0.05).4. The results of RANTESAt the same time point, RANTES concentration with the stocks of red blood cellsupernatant time increased (P <0.05) in red blood cell supernatant of the same stock.At6h,24h the release of RANTES in no difference (P>0.05), but with3h comparedto the significant differences (P <0.05). The same time point, RANTES concentrationwith LPS concentration in the solution increases (P <0.05).5. The results of IL-1βThe release of IL-1β in the7d supernatant stimulation, extended with theretention time increased (P <0.05);21d,35d supernatant to stimulate,3h,6h at the IL-1β release is not extended over time to increase (P>0.05), but in the24h at theIL-1β release increased with time (P <0.05). The same time point, IL-1β release withthe extension preserve the red blood cell supernatant increased (P <0.05). In the sametime, the concentration of IL-1β with LPS concentration in the solution increases (P<0.05).6. The results of CD62PIn7d supernatant stimulation, the levels of CD62P expression in6h,24hDepartment has yet to extend the time increase (P>0.05);21d,35d supernatant tostimulate the levels of CD62P expression with time increased (P <0.05). On the samepoint in time, CD62P expression with LPS concentration in the solution increases (P<0.05).7. The results of Platelet aggregation rateOn the same point in time, platelet aggregation with the stock red blood cellsupernatant time and lower (P <0.05), in the same inventory in the red blood cellsupernatant, platelet aggregation with the stock red blood cell supernatant time andlower (P <0.05);at the same time, platelet aggregation with the solution concentrationof LPS increase reduced (P <0.05).8. The results of Platelet hypotonic shockAt the same point in time, the rate of platelet hypotonic shock with the stock redblood cell supernatant time to extend the lower (P <0.05), decreased in the same stockin red blood cell supernatant, platelet hypotonic shock rate with the stock red cellsupernatant time extension, but not statistically significant (P>0.05); the same pointin time, the rate of platelet hypotonic shock solution, the increase of the concentrationof LPS decreased (P <0.05).9. The results of AnnexinVStatistical results at each time point and stimulate red blood cell supernatant wereno significant differences (P>0.05).10. The results of TNF-αTNF-α was not detected in all samples.Conclusion1. A suspended red blood cell retention time is longer the supernatant to stimulateplatelet, the stronger the ability to produce proinflammatory cytokines.2. Spended erythrocyte save time longer, the supernatant of platelet aggregationgreater the impact. 3. solution, the greater the concentration of LPS, platelets are more easilyactivated and produce proinflammatory cytokines.