The Effect Of SP And NGF On Activity Of Natural Killer Cells In Peripheral Blood Of Normal Human

PrefaceWhen peoples observed that the effects of stress on immune function in human ,the stress induce immune function low and the inhibit activity of natural killer cell , the quantity of leukomonocyte and lymphomonocyte also decrease . Neuroendocrine system could regulation organism immune function, while classics neurotransmitter,neuropeptide are important regulatory substance that affect on immune competent cell and other cells in directly . Scholar report ever,sp can enhancement proliferation cell of B production antibody , moreover incentive lymphomonocyte production cytokine participate in immune regulation effect. Literature indicate;no matter incubation in vitro or dosage in vivo . Sp can enhance activity of natural killer cell of leukomonocyte in small intestine epithelium , then also can enhance activity of spleen cell by dosage in vivo. Further Hart R observed sp can enhancement secretion of IgM and IFN of lymphomonocyte in periphery blood . From above data : sp is important regulatory peptide in corpore , and also adjust immune function.Natural killer cell is important cell colony . it almost participate in full adjusting process about occurrence, development, effective of immune system, at present, people study natural killer cell less than T cell and B cell . While the integrated report about NK cell act on role and function in neuroendocrine - immune network are very scarce . Hellstrand discovered if natural killer cell pre-treatment by low level adnephrin . It can raise activity of natural killer cell . But if 10 ^-6M density adnephrin incubation together with natural killer cell ,then activity of natural killer cell is very low . So NK is very sensitive to stressor.NGF and SP sibling polypeptide substance . Many data indicated : both have visible adjustment effect to immune function . Nevertheless the report aboutthe two affect to activity of mononuclearcell in periphery blood of human in vitro is not find . This experiment study the effect of NGF and SP on activity of NK cell in periphery blood of human , to expect approach the two to organism immune adjust effect .Materials and methodTake exponential phase of growth k562 cell , with no Ca2 + Mg2 + Hank's washings twice . trypanblau blue - stain . Sure to cells motility rate >95%. after viable count , with 1% BSA - RPMI -1640 take cell density adjust 1 105cell suspension reserve .Vein obtain blood 100ml , heparin sodium anticoagulation , use D - Hank s dilute three times , stratum super stratum leukomonocyte , 2000r/min centrif-ugalization 20min , take suction white coat cell . With no Ca2 + Mg2 + Hank's centrifugalization 1200r/min lOmin , washings twice, suspension in RPMI -1640 , trypanblau blue -stain . Sure to cells motility rate >95%. After viable count , with 1% BSA - RPMI - 1640 take cell density adjust needed cell suspension reserve at last .Use 96 shadow mask , add SP and NGF. take shadow mask of postvaccinal in incubate box of 37*05% CO2 4 hours. Centrifugalization 1200r/min lOmin. Every hollow take 0. lml supernatant to other 96 shadow mask , every hollow add LDH substrate mixed liquor 0. lml . Away from light 10-15 min , then every hollow add lmol/1 citric acid stop buffer 30jxl , enzyme scale appearance detect optical density , calculate activity of NK cell . All data are expressd as mean ± SD, significance of differences was determined by t test.ResultsWhen potency target ratio is 100:1 ,2 g/ml or 8 g/ml SP have inhibitory action on activity of NK cell ( P < 0. 05). When potency target ratio is 50:1 ,8 g/ml SP have inhibitory action on activity of NK cell( P <0. 05) . When potency target ratio is 25:1, 4 g/ml SP have enhancement property on activity of NKceU(P<0.05) .When potency target ratio is 100:1 and 25 :1, 40ng/ml NGF have inhibitory action on activity of NK cell (P <0. 05).DiscussionNK cell is organism natural existing anti - tumor cell . It active high or low with disease occurrence ^ development ^turnover have intimate relation . In this experiment , when potency target ratio is 100:1 , 2|xg/ml or 8jxg/ml SP have inhibitory action on activity of NK cell (P <0.05) . When potency target ratio is 50:1 , 8|xg/ml SP have inhibitory action on activity of NK cell( P < 0.05) . When potency target ratio is 25:1, 4 g/ml SP have enhancement property on activity of NK cell( P <0.05) . It indicate that adjustmen of SP to activity of NK cell with it dose close correlation . from above - mentioned , three aspect to analyze as follow: ( 1) Recently , due to use radio - immunity receptor assay meridian syndrome have SP receptor on immunocell . SP through it receptor NK -1 N NK -2NNK -3 receptor linking and educe biological effect . Because SP with NK - 1 receptor binding capability maximum , so NK - 1 receptor called SP receptor . SP educe physio - effect main path is by NK - 1 receptor . SP may incorporate with NK -1 receptor on the NK cell envelope , inhibit activity of NK cell . (2) SP urge PBMC production IFN - -y , IFN - y can increasing activity of NK cell . but if fore treatment with SP , then activity of NK cell depress , part neutralization IFN - -/enhancement property . This possible that SP linking with much cell surface receptor , membrane structure change obviously , to hinder IFN - y on NK, or because SP incentive macrophage create PG more negative growth factor , inhibit activity of NK cell . (3 ) Natural killer cell express two konds different receptor : One kind is killer activatory receptor;other kind is killer inhibitory receptor . With regards to natural killer cell two receptor regulation NK lethal effect mechanism yet not clear . At present , have many hypothesis , among the total, one receptive is : KIR on the NK cell surface col - with self MHC -1 , production restrain signal , this signal act main guide inside cell , it can block kill signal transmission . When SP urge PBMC production IFN - y ^TNF-aNIL-2 cytokine , under these cytokine incentive , induction MHC -I increase , then KIR mediated depressant effect hold main guide , so activity of NK cell be restrained.In experiment , when potency target ratio is 100;1 and 25: 1, 40ng/ml NGF have inhibitory action on activity of NK cell (P < 0. 05 ) . 40ng/ml may that NGF bring into full play max biological activity density . to guess it mechanism : ( 1) NGFR distribution all the nerve cell , but some non nerve cell envelope has NGFR . Target cell of NGF exist high avidity and low avidity , NGF may combine together NGFR on NK , internalization conveying pericaryon , then together with nuclear receptor produces depressive effect on target cell . But NGF whether combine with high avidity or low avidity still in research . (2 ) The cell envelope of NK exist IL -2R , IL -2 interaction with IL -2R , irritate NK cell multiplication^ differentiationN boost lethal effect . So NK corpuscular activity depend on continue existing small amounts IL - 2 in vivo . NGF could stimulate leukomonocyte proliferation and high express IL -2R . But if density of IL - 2R too much , then IL - 2R be act block receptor together with IL - 2 restrain immune function of target cell.NGF and SP belong to polypeptide substance . With regard to the effect of SP^NGF on activity of natural killer celUas well as adjust immune function , and produce a marked effect at;Immun physiology and immunopathogenesis is generally accepted . but it regulation trend and internal mechanism , the result of report not with one accord . Some consider strengthen effect , some consider depressant effect . The result whole and ex vivo are not coincidence . Cause to phenomenon are two;One explain that the accommodation of neuropeptide and neurotransmitter immune function is very complex , different function and different condition induce different effect . For example , relate to experimenter adoption densityAspeciesNorganization^cell type . another aspect illustrate that study not deep enough at present.Sum up above , body and mind stimulation act on nervous system , impact immune function by nervous in direct , and accommodate immune function by release endocrine krinin or cytokine . SP and NGF are neuroendocrine and immune system chemi - language . SP^NGF and its fragment induce effect is verycomplex and diversity in different conditions . To study advanced that the mechanism of SP^NGF on neuroendocrine -immune network , and to obtain renewal understand . another aspect is master new development of neuroendocrine immu-nological regulation network about clinical disease . Expect to help of clinical prevention and cure.Conclusion1. SP has depressant effect on peripheral blood mononuclear cell in human . when potency target ratio is 100:1 ,2 g/ml or 8 g/ml SP have inhibitory action on activity of NK cell , compare to control (P <0.05) . when potency target ratio is 50;1 ,8 g/ml SP have inhibitory action on activity of NK cell, compare to control ( P < 0.05 ) .2. NGF has depressant effect on peripheral blood mononuclear cell in human . when potency target ratio is 100:1 and 25:1, 40ng/ml NGF have inhibitory action on activity of NK cell , compare to control (P <0.05).