Protective Effects Of Vaporizing Inhaled Perfluorocabon On The Organs Outside Lung Of Acute Lung Injury Pig

Objection ventilator associated lung injury, which caused outside lung organs damage has already become the problem that could not be avoided in the ventilator use. Many protective ventilated strategies arise at the historic moment. The perfluorocarbon vaporized inspiration treating acute lung damage has recently aroused people's interest. The effect of this kind ventilated strategy on the internal organs not yet to see the report to the lung outside, this research is for the purpose of understanding the protective effect of perfluorocarbon vaporized inspiration on the internal organs outside lung of acute lung injured pig model.Methods 16 piglets are randomly assigned to 2 groups, (1) Vaporized-PFC group, (2) CMV-Control group. After successful anaesthesia, tracheal intubation and CMV (volume control, VT 8ml/kg, f 30/min, FiO2 0.6,1:E 1:1) are performed. When common conditions are stable, animal model of ARDS is induced by detergent infusing through tracheal intubation. After a stable animal model of ARDS established, group (1) received 2 hours CMV and vaporized PFC inhaling through a humidifier, and CMV continued 6 hours after therapy, group (2) received 8 hours CMV. Blood gas analysis, lung mechanic (PIP, PPL, VTE, Cst) and hemodynamic (MAP, HR) parameters are obtained at the moment of baseline, ARDS animal model succeed and 15-min intervals during 2 hours therapy and 30-min intervals during 6 hours post-therapy period. As the results shown, at the moment ARDS animal model is established.Bacilliculture, biochemical marker and cytokine density : Under the aseptic condition are the liver, spleen, the pancreas, the kidney, the small intestine and periphery the lymph node organization taken out and made into homogenate. The homogenate is put into 35°C hatch box to foster for 24 hours. The colony size shape and counts are observed. Radioimmune assay measures TNF- α ,ET-1,IL-8and TGF- 3 concentration in blood plasma. 7,600 Hitachi completely automatic biochemistry analyzer examine blood plasma hepatorenal pancreas function target density.Outside lung internal organs histiocyte morphology after HE dyes is fathomed by optical microscope. The apoptosis index determination by TUNEL: stochastically takes 5 high time of fields of vision (40X/0.65 objective lens), divides in the field of vision by the masculine cell number the cell total to express apoptosis index, by percentage expression. Transmission electron microscope examines cell ultrastructura variance. The immunohistochemical method examines BAX, bcl-2, COX-2, TGF- beta 1 protein expression. Semiquantitative RT-PCR law determines BAX, bcl-2,COX-2,TGF- beta 1 mRNA of outside lung organs.Results1. liver, spleen, kidney, pancreatic gland and small intestine bacilliculture resultThe liver and spleen organization refining culture of bacteria colony number in CMV and the VMV group is 0, the kidney, the pancreatic gland and the small intestine and periphery lymph node in CMV and the VMV group raises the bacterium colony, the VMV group compares the CMV group obviously to reduce2. liver, kidney, pancreatic function resultWhen acute lung damage model success, measured blood plasma foundation value AST, ALT, LDH, AMY and Cr do not have the remarkable difference between CMV and the VMV(P>0.05). After the vaporization 2h, or 8h, blood plasma AST, ALT, LDH, AMY have the remarkable difference between CMV and the VMV (P<0.05), the VMV group various targets compare the CMV group remarkably to reduce.3. Blood plasma Inflammatory factor density resultWhen acute lung damage model success, measured blood plasma foundationvalue TNF-a , ET-l,IL-8 and TGF-P do not have the remarkable difference between CMV and the VMV (P>0.05). After the vaporization 2h, or 8h, blood plasma TNF- a , ET-1, IL-8 and TGF- 3 have the remarkable difference between CMV and the VMV (i^O.05), the VMV group various targets compare the CMV group remarkably to reduce.4.Optical microscope observed resultsVMV group is that liver congestion degree obviously is lower, liver cell dropsy is milder, in sinus hepaticus seeps out lighter, the leaflet structure is completer, the inflammatory cell infiltrate is lower than CMV group. The VMV group red and white pulp dividing line is clear, the splenic corpuscle germinal center is not very obvious, compared with CMV group. Apoptosis and necrosis degree of kidney proximal tubule epithelial cell is lower in VMV than CMV group. VMV group pancreatic gland acinus necrosis reduces, apoptosis increase, glandular duct structure integrity, compared with CMV group. Small intestine in the VMV group appears subepithelial space to expand, the gland mildly suffers injury and the blood capillary hyperemia, apoptosis cell obviously to reduce, compared with CMV group.5.Transmission electron microscope observed resultsThe cell necrosis and apoptosis of liver, spleen, renal tubule and intestine villi in VMV group are more lightly than CMV group. Pancreatic acinar cell apoptosis in VMV group is more, necrosis lower, than CMV group.6. Apoptosis percentage resultsTUNEL masculine cell presents the varying degree brown. The liver masculine cell mainly distributes in the central vein encircling zone, majority of masculine cell amnion integrity. The nucleus changes slightly, the karyoplasm proportion reduces. Some cells shrink firmly, obviously changes slightly, the cytoblastema basically vanishes, assumes apoptosis body.The spleen masculine cell disperses in distributes in white pulp, splenic sinus. In the VMV treatment group liver, and spleen, the masculine cell number is morethan the CMV group.The CMV group the apoptosis index to be high, the apoptosis cell to be most is proximal and distal convoluted tubule epithelial cell. The renal glomerulus has not seen apoptosis cell.The VMV group compares CMV group apoptosis rate obviously to reduce (P < 0. 05).The CMV group pancreatic gland few disperses in acinus necrosis focus, apoptosis cell to be few;The VMV group compares the CMV group apoptosis cell to increase, necrosis to reduce.The CMV group appears the down epithelial cell to fall off, the gland seriously suffers injury, apoptosis cell many, the above changes are especially remarkable in the down peak. The VMV group appears under the epidermis the gap to expand, the gland mildly suffers injury and the blood capillary hyperemia, apoptosis cell obviously to reduce.7. BAX, bcl-2,COX-2, TGF- beta 1 protein expression resultsThe VMV group is compared with the CMV group that the liver, the kidney and small intestine BAX reduces, Bcl-2 elevates, while the pancreatic gland BAX elevates,Bcl-2 reduces (P <0.05). Spleen BAX, bcl-2 between two groups have no discrepancies. The VMV group is compared with the CMV group that small intestine COX-2 remarkably elevates (P<0.05), the kidney TGF- 3 1 significantly reduces (P<0.05). The protein expression of BAX, bcl-2,COX-2, TGF- beta 1 is finally consistent with their corresponding mRNA result.8. BAX, bcl-2,COX-2, TGF- beta 1 mRNA resultThe VMV group is compared with the CMV group that the liver, the kidney and small intestine BAX mRNA reduces, Bcl-2 mRNA elevates, while the pancreatic gland BAX mRNA elevates,Bcl-2 mRNA reduces (P <0.05). Spleen BAX mRNA, bcl-2 mRNA between two groups have no discrepancies. The VMV group is compared with the CMV group that small intestine COX-2 mRNA remarkably elevates (P<0.05), the kidney TGF- 3 1 mRNA significantly reduces (P<0.05).Conclusion1 .The conventional machanical ventilation may cause the function of outside lung internal organs in the ALI pig out of balance, the small intestine barrier function being reduced, bacterium shifting.2.The perfluorocarbon vaporized inhaled possibly alleviates the function failure of outside lung internal organs, the small intestine barrier being damaged, bacterium shifting.3.Through reducling the cell apoptosis, the vaporized ventilation possibly alleviates outside lung internal organs function out of balance, the small intestine barrier function reduced, bacterium shifting.4.The ALI small pig liver, the pancreas, the kidney, the small intestine cell apoptosis is possibly regulated by BAX, the bcl-2.5.The cell apoptosis of outside lung organs is individedly regulated by COX-2 or TGF-P1 and so on.6.Cell factor TNF, IL-8, ET-1 and TGF-Pis possible to induce outside lung internal organs cell apoptosis in the ALI pig.7.Possibly through reducing the inflammation factor level in the blood, the vaporized ventilation adjust the apoptosis gene to regulae apoptosis.