The Effects Of Infrasound On Human Peripheral Blood Mononuclear Cell

Backgrounds: Infrasound (10^-5～20 Hz) is a mechanical vibrational wave, which is below the audibility range of the human ear (20 Hz to 22 kHz).With the development of science, the potential use of infrasound in clinical diagnosis and treatment has been regarded increasingly. Some experts have claimed Infrasound can enhance the immunity function and the morbid resistance. But until now there are few research and almost no reports about the mechanism.Human immune system can fight infection and adapt to intruders. And human peripheral blood mononuclear cell (PBMC) is a critical component in immune system, to be more exact, is a blood cell having a round nucleus, such as a lymphocyte or a monocyte. As a sophisticated method, PBMC can be extracted from whole blood using ficoll, a hydrophilic polysaccharide that separates layers of blood, with monocytes and PBMC forming a buffy coat under a layer of plasma. This buffy coat contains the PBMCs. At present, to measure body immune function PBMC is widely used in research and clinical every day.Objective: In this thesis, we investigate some effects of infrasound on PBMC. Concretely, we will study infrasound on the active of cells, the ultrastructure of PBMC, the cell cycle and apoptosis, to provide an experimental basis for effects of infrasound on immune system and apply infrasound design of parameter. Hypotheses of study: Infrasound treatment in the experiment might increase the active of human peripheral blood cell. Similarly, the effects of infrasound on the apoptosis of PBMC and the ultrastructural change of PBMC can be found.Methods: we extracted PMBC from whole human blood using ficoll, The cell viability would be showed by trypan blue staining .And we used INFRASOUND 8^TM , a therapeutic infrasound generator, to treat the vitro cultured PMBC (treatment groups). The cells of control groups were exposed to air. While the bank group were cultivated in incubated tank with 5% CO2 and saturated humidity all through, do not be exposed to infrasound or air .The proliferation of the cells would be measured respectively at 15 min, 30 min, 60 min, 90 min and 120 min After the treatment all the cells were cultivated for 24 h and 48 h separately. Then the cell multiplication measured by MTT assay, and the development of cell apoptosis caused by different management groups was examined by flow cytometry analyses (FCM). The scanning electron microscopy (SEM) and transmission electron microscopy (TEM) would be used for the test. The data was conducted and analyzed with spssl3.0. And statistical evaluation was done by repeated measure ANOVA. Post hoc analysis was performed by using LSD post hoc analysisResults:1. The results of trypan blue assay showed that the rates of survival PBMC in all groups were above 95% .2. MTT assay: OD counts of PBMC in infrasound groups were no differences from their control groups (P>0.05), but that of the infrasound groups showed increasing with the time of infrasound treatment. OD counts of PBMC in control groups did not display such a tendency.3. Flow cytometry analysis: Percentages of apoptosis cell and other cells had no significant difference in all groups (P>0.05).4. SEM: The PBMC treated by infrasound exposure for 120 min and then cultivated for 48 h showed that the prominent or micro-floss of the membrane and the figure were almost same as the cell of blank group. But the cell of control group showed that the prominent or micro-floss of the membrane become shorten or decreased, the abnormal figure of the cell increased.5. TEM: The cells in the black group were in a good condition, with normal mitochondria and endoplasmic reticulum in the cytoplasm and with plenty of follicles on their membrane. While the deteriorated microvillis, irregularin shape and homogenizing cell nuclear were showed in the cells in the control group. Even the membrane of some nucleus were broken and autophagic vacuoles took place of organelles. However compared with the cells in the black group and the control group, the condition of the cells in the infrasound group were in the middle. Clear but irregular-organized nuclear membrane and slightly swelling of mitochondria were showed in the cells in the infrasound group. And the microvillis on their membrane decreased a little.Conclusion:Infrasound treatment in the experiment might increase the active of human peripheral blood cell, but the influence on apoptosis of PBMC was not obvious. The membrane of PBMC would be affected directly by infrasound. Infrasound therapy might prevent natural aging of PBMC and damages from environment. But the effect were not evidence.Summary:Infrasound treatment in the experiment might increase the active of human peripheral blood mononuclear cells, but the influence on apoptosis of mononuclear cells was not obvious. The membrane of mononuclear cells would be affected directly by infrasound. Human immune system might be affected by infrasound. And further research of other exact mechanism are expected.