Empirical Study For Gene Transfection Of Interleukin-10 To Inhibit Ventilator-induced Lung Injury

Objective:Mechanical ventilation(MV)is widely used in anesthesiology,critical care medicine,emergency,IntMED and so on,but at the same time,ventilator-induced lung injury effects prognosis for severe patients.The experiment takes molecular biology technique, gene transfect IL-10 to lung tissue to inhibit the releasing,of cytokine and phlegmonosis medium in VILI.The experiment is to evaluate the treatment from observing TNF-α,IL-1β,IL-8,IL-10 and changes of morphology in lung tissue.At last,it will offer a theory for clinic to prevent and cure VILI.Methods:54 male SD(Sprague Dawley)rats were randomly divided into 9 groups,6 every group.Group A:control group(have no deal);Group L:low airway pressure(15cmH₂O),it divided to L₂ and L₄,depending on time of mechanical vetilation; Group H:high airway pressure(25cmH₂O),it divided to H₂ and H₄,depending on time of mechanical vetilation;Group B-.Ad-Blank gene transfection(25cmH₂O),it divided to B₂ and B₄,depending on time of mechanical vetilation;Group T:ADS-h-IL-10 gene transfection(25cmH₂O),it divided to T₂ and T₄,depending on time of mechanical vetilation. The experimental rats were injected with 10%chloraldurat(0.3g/kg)at abdominal cavity,and then cut-off trachea at the middle of neck.After inserting tracheal catheter and connecting breathing machine,open the pectoral cavity at lefe side between 3,4 cervical ribs,inject agentia from right ventricle.The agentia is respectively:Lactic Acid for groupL,H;Ad-Blank for group B;AD5-h-IL-10 for group T(all groups is 0.1ml).Close the pectoral cavity,removal tracheal catheter and intramuscular injection penicillin 10⁵U to prevent infection.After 48 hours,recut-off trachea and insert tracheal catheter to give a persistent mechanical ventilation.In the process,boost 10%chloraldurat rely on your need.For the reservative time,open the abdominal cavity,collect blood from abdominal aorta. Then centrifugalization the blood at rate of 3000r/min for 15min,collect supernatant fluid to preserced in refrigerator(-80℃).Take lung tissue to detect pathology change.At last,detet the contents of TNF-α,IL-1β,IL-8,IL-10 in blood serum.All the data were analyzed by SPSS11.5 statistics software,P value＜0.05 was considered to have statistical significance. Result:(1)Light microscope HE:Group A:pulmonary alveoli are as normal;Group L:there is a little effusion in pulmonary alveoli and some phlegmonosis cell besides the blood vessel;the extent aggregate with mechanical ventilate time;Group H and B is serious than L and T is lessen;(2)Electron microscope:Group A:structure of cellâ… ,â…¡and endotheliocyte are as integrity as normal;Group L,H,B,T:bioblast swell,ablate and its crista disappear,endoplasmic broaden,cell nucleus enlarge,bland between cell nucleus and cell-substane broaden,caryotin bad-distribute,lamellar body swell,microvilli decrease with different degree;the change is not obvious;compared with group H and B,T is lessen in some entent;(3)Inflammatory factors from blood serum:â‘ Content of IL-1β:Group L,H,B,T is higher than group A(P＜0.01);content of IL-1βhighten with time prolong(P＜0.01);with the same ventilation time,H is higher than L(P＜0.01)and T is lower than B(P＜0.05);â‘¡Content of TNF-a:Group L,H,B,T is higher than group A(P＜0.01);content of TNF-ahighten with time prolong(P＜0.01);with the same MV time,group H is obvious ly higher than group L(P＜0.01)and group T is lower than group B(P＜0.05 or 0.01);â‘¢Content of IL-8:Group L,H,B,T is higher than group A(P＜0.01); content of IL-8 highten with time prolong(P＜0.01);with the same MV time,group H is obviously higher than group L(P＜0.01)and group T is lower than group B(P＜0.05 or 0.01);â‘£Content of IL-10:Group L,H,B,T is higher than group A(P＜0.01);content of IL-10 highten with time prolong(P＜0.01);with the same MV time,group H is obviously higher than group L(P＜0.05 or 0.01)and group T is higher than group B(P＜0.01).Conclusioa:(1)VILI rats intertitium of lung edema,widen and many phlegmonosis cell collect,pulmonary alveoli bloodloss and form hyaline membrane in the surface;(2)At the same airway pressure,content of TNF-α,IL-1β,IL-8,IL-10 enhance with the time prolong;(3)With the same time of mechanical ventilation,content of TNF-α,IL-1β,IL-8,IL-10 enhance with the highten of airway pressure;(4)IL-10 gene tramsfection can amendment morphology change of lung tissue,lessen edema and infiltration of phlegmonosis cell;(5)IL-10 gene transfection down regulation the expression of TNF-α,IL-1β,IL-8 in blood serum and up regulation IL-10 to enhance the ability to anti-phlegmonosis,at last to lessen injury of lung. Objtctive:Approach influence on lung shunt fraction from changes of body position during general anaesthesia.Observing the influence of shenfu injection on lung shunt fraction,and comprehend the function of shenfu injection on protection of lung,at last provide an evidence for the use of shenfu injection in clinic.Methods:24 patients(ASAâ… -â…¡)for back bone selective operation were randomly divided into 2 groups,12 every group.The experimental group give shenfu injection intravenously,with the dose of 1ml/kg.Monitoring BP,HR,SpO₂ after the patients in.All the patients use the same drug in induction of anaesthesia,including midazolam0.04mg/kg,fentanyl 3-4ug/kg,propofol 1-1.5mg/kg,atracurium 0.6mg/kg or vecuronium 0.1mg/kg.Aftering intubate the catheter,set the V_T 8-9ml/kg,rate is 12/min.Punct the left radial artery and right internal jugular vein to moitor BP and CVP.We use sevoflurane and propofol to maintenance anesthesia.Compared the BP,HR,SpO₂,P_ETCO₂,Raw and C_dynat T₁(30min of supine position)and T₂(30min of prone).Cull blood from radial artery at two time point and do blood gas analysis.Compare the change of PaO₂ and PaCO₂.Calculate lung shunt fraction from the formula:Qs/Qt:=A-aDO2×0.0031/A-aDO2×0.0031+5 A-aDO2=(760-age)×FiO2-(PaCO2×1/0.8)-PaO2(FiO2为1.0).All the data were analyzed by SPSS11.5 statistics soflware,P value＜0.05 was considered to have statistical significance.Result:(1)Patients in two groups have no difference in age and sex.;(2)SBP in patients of control group have significant difference at T₁,T₂(P＜0.01),DBP have no difference at T₁,T₂;(3)PaO₂,PaCO₂ and P_ETCO₂ have no difference in two groups at T₁,T₂(P＞0.05);(4)Cydn in two groups at T₁ time point have difference(P＜0.05)and T₂ have no difference.In the same group,there is no difference at two time points(P＞0.05);(5)Lung shunt fraction present a tendency of rising,but have no difference in statistics(P＞0.05).Conclusion:We can conclude that from the study,lung shunt fraction present a tendency of rising,but have no difference in statistics when the patients from supine position to prone position;shenfu injection has a function in sustaining the blood pressure and lung compliance during anaesthesia,but have no significant influrance on lung shunt fraction to health adult.