Changes Of Heme Oxygenase-1 And Apoptosis As Well As The Relationship Between Them Following Carbon Monoxide Poisoning In Mice

Background: The Carbon monoxide poisoning is common in life and production of toxic diseases, severely endangering humanity's health and life.The acute carbon monoxide poison patient passes through the prompt rescue treatment, mostly may be restored to health, but has a part of patient to alleviate, consciousness after the toxicant symptom restores normally, probably about 2-60 days experience normal or the basic normal interim period, can appear a series of brain illness once more the symptom, known as carbon monoxide poisoning delayed encephalopathy (DEACMP). Uemura and other reports,the patient died of acute CO poisonin gautopsy found that the cortex, basal ganglia, hippocampus, thalamus and cerebellum apoptosis and death coexistence, CA2 apoptosis in the hippocampus area more obvious [1].The brain cells necrosis occurred mainly in acute CO poisoning caused by a serious injury, disease and inflammatory response to injury for Features [2], and DEACMP is caused by acute CO poisoning Experience a "normal" a period, to speculate DEACMP symptoms of CO poisoning and the emergence of apoptosis of nerve cells continue to take place is contacted together[3].At present ,The Heme oxygenase (HO) biological characteristics have a more clear understanding of its presence in the body.It has three kinds of existence forms, namely HO-1, HO-2, HO-3. HO-1 which was induced. A variety of physical and chemical factors stimulated increased expression of HO-1. In the normal brain tissue, it mainly expresses the HO-2, HO-1 expression of little or no expression . Some literatures report [4,5]in animal experiments HO-1 in hypoxic ischemic encephalopathy associated with apoptosis.Objective: study the carbon monoxide poisoning at different time points in mice brain Heme oxygenase-1 expression changes;Detect carbon monoxide poisoning at different time points in mice brain nerve cell apoptosis and related gene expression situation; Discuss relations of heme oxygenase-1 and brain cell apoptosis changes. Further speculate mechanism of encephalopathy of carbon monoxide poisoning. To further explore new ways of clinical treatment and provide a theoretical basis.Methods: Male mice, 18 ~ 22 g, were divided into CO poisoning group and air control group . CO poisoning group will be injected by using the weight (150ml/kg) every six hours for spacing fourth of high purity (99.9%) of CO gas in mice exposed to modified model. Air control Group will be injected by using of body weight (150ml/1kg) every six hours for spacing fourth of air. The hippocampus brain tissue pathological form of injury was observed by hematoxylin - Yihong staining . HO-1, Bcl-2, Bax in the level of protein expression were observed by SABC immunohistochemical method. HO-1 in the level of protein expression was observed by using Western blot. Cell apopsis was detected by flow cytometry. Under the 400 times the light microscope, counting an average of five outlook positive cells, all datas were expressed by mean±sd .statistical analysis software of SPSS13.0 t-test was employed.Results: 1,light microscope hippocampus of the brain tissue pathological changes: (1) the air in the control group each time point brain slices were no obvious changes. (2) CO poisoning Group 1 day foreseable edema nerve cells, slightly reducing the number of cells, cells with desalination; 3 days in addition to the appeal, the visible part of cell necrosis, nuclear enrichment, expansion of space ; 5 days of this phenomenon further clear that the hippocampus occur in the large number of cell necrosis, cell edema increase. In 21 days cell edema reduce,but the visible part of cell necrosis.2,Immunohistochemical method HO-1 protein expression: air control group less positive cells; CO poisoning group more positive cells in the hippocampus, HO-1 expression increased in 1 day(52.75±6.5)(p<0.01), reached the peak(98.0±8.64) (p<0.01) in 3 days, 5 days decreased(73.0±8.6) (p<0.01), in 21 days(35.5±2.08)still higher than that in control group. (p<0.01). 3,Western blot method HO-1 protein expression: air control group to express less, CO poisoning group in 1 day HO-1 expression increased(1.14±0.43) (p<0.01), reached the peak(2.91±1.07) (p<0.01) . 3 days declining slightly, still higher than that in control group. (p<0.01) . And agreement with immunohistochemical method measured HO-1 protein expression.4,apoptosis: air control group at different time points less apoptosis, CO poisoning group the number of apoptosis increased in 12 h(21.95±24) (p<0.01) , in 5 days apoptosis reached the peak(34.27±4.19) (p<0.01), in 15 days apoptosis reduced(16.6±0.78) (p<0.01), In 21days(9.25±1.16)still higher than that in control group (p<0.01).5,apoptosis related protein expression: Bax expression increased after the poisoning, reached the peak in 3 days(34.5±0.58) (p<0.01), after it declining slightly. Bcl-2 expression increased after the poisoning, 3 days reduced to a minimum(12.5±3.69) (p<0.01), Bax/Bcl-2 the ratio of increase, it is the largest in 3 days.Conclusion: 1,normal mice hippocampus may be a small amount of HO-1 expression, CO poisoning in mice HO-1 after two methods of expression increased significantly.2,the normal mice hippocampus can be a very small amount of apoptosis cells, CO poisoning in mice hippocampus apoptosis cells have markedly increased, and with HO-1 expression of the agreement.3,which strongly suggested that HO-1 is asoociated with the mechanisms of brain damage caused by carbon monoxide poisoning.HO-1 makes change to protein expression related to apoptosis.HO-1 is candidate for neuronal apoptosis. Then for the clinical treatment of new ideas.