| PrefaceSubstance P is excitatory neuropeptides synthesised in neural and glial cells of the human central and peripheral nervous syste,one of the earliest neuropeptide,and widely distributed in the fine primary afferent nerve fibers.Physical and mental stimulation to the nervous system,can affect the direct effect of the immune system function,or through the release of hormones or other cytokines to regulate immune function,and the classical neurotransmitters,neuropeptide is a direct role in immune cells involved in immune response and other important regulators of the cell.Former reseach have revealed that effect of SP via combination with its receptor and morphology data that many cell surrface express sp receptor provide the evidence of sp involved in immune response and other important regulators of the cell.Some immune cell also produce SP which worked by autocrine or paracrineNK cells constitute an important component of the innate immune system,played an important role in the anti-infective,anti-tumor,immunoregulation and hematopoietic system.NK cells exert cell-mediated cytotoxicity and act to regulate innate and acquired immune responses through the release of various cytokines(such as IFN,GM-CSF and TNF) and chemokines.NK cell activity is regulated by an array of membranebond inhibitory and activating receptor.Yet mechanisms of substance P(SP) function on NK cells is still ambiguityHere,we investigated if SP boosts proliferation and cytotoxicity of NK1 receptor specific antagon pretreated natural killer cells,and observed the effect of substance P (SP)on the expression of NK1R to study the effect of SP on NK function and the possible mechanism of action.Materials and Methods1.Experimental group were pretreated with NK1 receptor specific antagon. Experimental group and control group respectively cultured and treated with SP in different concentration for 24h.2.The proliferation of human NK cells(NK92-MI) treated with SP(10-4mol/L 10-6mol/L,10-8mol/L,,10-10mol/L,10-12mol/L,10-14mol/L) was detected by MTT method and FACS.3.the cytotoxicity of NK92MI was checked through MTT assay.4.RT-PCR was used for the rnRNA expression of NK1R and NK2R5.FACS was applied to measure the expression of NK1RResults1.Group no pretreated with 10-7M NK1 receptor specific antagon with SP at 10-10M.～10-6M significantly promotes the proliferation of NK92MI compared to control.2.SP at 10-14M～10-8M up-regulate the cytotoxicity of NK1 receptor specific antagon pretreated natural killer cells and untreated cell.NK92-MI at 4:1,and the effect of SP cut down after the concentration up.two groups shows no difference.3.NK92MI co-cultured with SP for 24h,RT-PCR measured the mRNA level of the receptors NK1R.The results suggested that the mRNA expression of NK1R up-regulated with 10-10M.～10-6M4.FACS shows the membrane expression of NK1R,compare to the control,was upregulate at 10-10M.～10-6MConclusionsNK1 receptor is expressed on NK92MI cell.SP in a scope of concentration could promote NK1 receptor expression and the proliferation of NK92MI and strrong point in 10-10M.NK1 receptor specific antagon can completely inhibit this effect.SP in a scope of concentration could up-regulated cytotoxicity activity of NK92-MI;especially in 10-10M,10-8M,10-6M NK1 receptor specific antagon partly inhibit the cytotoxicity of NK92MI induced by sp,so we supposed that proliferation of NK92MI induced by sp is mediated by NK1 receptor and cytotoxicity maybe mediated by some other paths.
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