The Study On Ultraviolet Light In Combination With Ozone And Hypothermia's Effect Of Inducing Apoptosis In Human Peripheral Blood Lymphocytes

【Objective】Some scholars have applied ultraviolet light in combination with ozone and hyperthermia to promote apoptosis in human peripheral blood cells, and then injected the blood back into the donor. The apoptotic blood cells (mainly lymphocytes) induced the body to produce immune tolerance and anti-inflammatory response which led to the treatment of some immunologic or inflammatory diseases. In this study we observe the apoptosis induced by ultraviolet light, hyperthermia and ozone on human peripheral blood lymphocytes to establish a stable blood cell inducing system for basic and clinical research.【Methods】After inducing peripheral blood of healthy adults through ultraviolet light, hyperthermia, ozone, respectively and jointly, we separated peripheral blood lymphocytes by means of lymphocyte separation medium density gradient centrifugation. We studied the characteristics of apoptotic cells on morphological, molecular biological and cellular levels with Wright - Giemsa stain smear microscopy, DNA gel electrophoresis and flow cytometry analysis.【Results】The morphological characters of lymphocytes were tested by microscope. The groups of lymphocytes induced by ultraviolet light, hyperthermia and ozone, appeared typical morphological changes of apoptosis and DNA typical ladder type line spectrum. Each apoptosis situation was quantitative determined by flow cytometry. After ultraviolet irradiation (15W, at a distance of 5cm) for 10, 20, 30 min, the apoptosis rates were 12.03%, 16.98 %, 26.58%. After induction by ozone concentrations on 14.5, 29, 43.5μg/mL for 20 min, the apoptosis rates were 6.76%, 26.38%, 35.1%. After the high temperature at 40,42.5,45℃for 20 min, the apoptosis rates were 15.9%,26.83 %, 39.88%.After ultraviolet irradiation(15W, in the 5cm position) and the high temperature at 40,42.5,45℃for 20 min, the apoptosis rates were 26.3%, 36.3%, 54.5%. After ultraviolet irradiation unit hyperthermia at 42.5℃and Ozone concentrations on 14.5,29,43.5μg/mL for 20 min, the apoptosis rates were 49.6%, 76.2%, 67.8%. The lymphocyte apoptosis rates induced by ultraviolet light unit ozone and hyperthermia increased significantly. The apoptosis rates of lymphocytes increased in a dose and time-dependent fashion within a certain range, and reached the highest point after ultraviolet irradiation unit hyperthermia at 42.5℃and ozone concentrations on 29μg/mL for 20 min.【Conclusions】Ultraviolet light, hyperthermia and ozone could induce apoptosis of lymphocytes in peripheral blood respectively, and the apoptosis rates increased in a dose and time-dependent fashion. Ultraviolet irradiation unit hyperthermia or ultraviolet irradiation unit hyperthermia and ozone could induce apoptosis of lymphocytes in peripheral blood significantly. The apoptosis rates of lymphocytes reached the highest point and then dropped. However, necrosis rates continued rising.