| Objective: Establish hemorrhagic shock model by using Bama miniature pigs of environmental hypothermia, with the aim to investigate the protective effect of sevoflurane Pre-conditioning or Post-conditioning towards important organs such as heart, liver, kidney, brain, lungs, intestines etc.Method: 32 Bama miniature pigs(weighing 22±5kg, male or female unlimited) were randomly divided into four equal groups with every group 8 pigs: sham group of environmental hypothermia(group DS), hemorrhagic shock group of environmental hypothermia(group DHS), sevoflurane pre-conditioning group of environmental hypothermia(group DPre /Sev) and sevoflurane post-conditioning group of environmental hypothermia(group DPost / Sev). Propfol was administrated intravenously for tracheal intubation and mechanical ventilation. Right internal jugular vein and right femoral artery were cut opened and pulmonary artery flotation catheter.Bilateral femoral artery were cannulated and respectively for hemodynamics and core temperature monitoring and for blooding, or blood sampling. After waking up the pigs in each group were placed in an air-conditioner(set temperature at-15℃), the animals were exposed respectively 2% sevoflurane for 30 min before and after HS. group DS was only for catheterization, but not bloodletting treatment. All experiments conducted in air-conditioner(set temperature at-15℃) At the points before hemorrhagic shock(T0), 0.5h after hemorrhagic shock(T1), 1h after hemorrhagic shock(T2), 1.5h after hemorrhagic shock(T3), 2h after hemorrhagic shock(T4), 3h after hemorrhagic shock(T5) and 4h after hemorrhagic shock(T6), mean arterial pressure(MAP) and heart rate(HR) were monitored and recorded for every group, and blood sample was collect centrifuged and frozened for, detecting the serum concentration of IL-6、TNF-α and SOD. All the animals were observed to death and were sacrificed at 4h after the end of blood withdrawal. Blood samples were used for examining the biochemical parameter of heart liver and kidney( CK,CK-MB,ALT,AST,BUN,SCr). Water content of stomach and intestines were detected. The organs(heart, lung, liver, kidney, brain and intestines) Removed for HE staining to observe the pathological changes in different organs. Brain tissues of S100β was detected by Enzyme-linked immunosorbent assay(ELISA) and I-FABP levels in intestine was detected by Western-blot.Results:1. The change of every group of pigs in MAP, HR and the core temperatureCompared with group DS, the MAP of group DHS, group DPre/Sev and group DPost/Sev were obviously decreased(P<0.05), HR was increased significantly(P<0.05). Compare with group DHS,group DPre/Sev and group DPost/Sev MAP were showed no obvious changed( P > 0.05), HR was obviously decreased(P<0.05).Compare with group DPre/Sev and group DPost/Sev, there was not significant difference(P>0.05). Within 4h,the core temperature of group DS and group DHS were downed from 38.75±0.71℃ and 38.67±0.75℃ to 37.24±0.61℃ and 34.48±0.47℃,there were obviously significant differences at T4(P<0.05). Compare with group DHS,group DPre/Sev and group DPost/Sev the core temperature were obviously reduced(P<0.05). there was not significant difference after T4(P>0.05).2. The change of inflammatory factor and oxygen free radicals2.1 Compared with group DS,group DHS, the TNF-α and IL-6 of secrum of group DPre/Sev and group DPost/Sev were increased at 1h after hemorrhagic shock and 1.5h after hemorrhagic shock(P<0.05).The TNF-α was reached the maximum at 2h after hemorrhagic shock,was decreased at 3h after hemorrhagic shock(P < 0.05) Compared with group DHS,TNF-α and IL-6 of secrum of group DPre/Sev and group DPost/Sev were decreased(P<0.05).Compare with group DPre/Sev and group DPost/Sev,there was not significantly different(P>0.05)2.2 Compared with group DS,SOD of secrum of group DHS,group DPre/Sev and group DPost/Sev were obviously decreased(P<0.05). Compared with group DHS,SOD of secrum of group DPre/Sev and group DPost/Sev were obviously increased(P<0.05).Compare with group DPre/Sev and group DPost/Sev,there was not significantly different(P>0.05)3.The effect on the vital organs3.1HeartCompared with group DS,biochemical parameter of heart( CK,CK-MB) of group DHS, group DPre/Sev and group DPost/Sev were increased( P<0.05).Compared with group DHS, biochemical parameter of heart( CK,CK-MB) of group DPre/Sev and group DPost/Sev were decreased(P<0.05).Compare with group DPre/Sev and group DPost/Sev,there was not significantly different(P>0.05).HE staining showed that no significant changed in myocardial cells of group DS. Group DHS inflammatory cell infiltration and swelling, necrosis of myocardial fibrosis, group DPre/Sev and group DPost/Sev of inflammatory cell infiltration in less damaged than that of the group DHS.3.2 LiverCompared with group DS,biochemical parameter of heart( ALT,AST) of group DHS,group DPre/Sev and group DPost/Sev were increased(P<0.05). Compared with group DHS,biochemical parameter of heart( ALT,AST) of group DPre/Sev and group DPost/Sev were decreased(P<0.05). Compare with group DPre/Sev and group DPost/Sev,there was not significantly different(P>0.05). HE staining showed that group DS liver was lobule structure completed,group DHS degeneration hepatic lobules, concentrated in liver cells, inflammatory cell infiltration, Group DPre/Sev and DPost/Sev of liver cells with mild edema, inflammatory cell infiltration inportal less damage than that of the Group DHS significantly reduced.3.3 KidneyCompared with group DS,biochemical parameter of heart( BUN,SCr) of group DHS,group DPre/Sev and group DPost/Sev were increased(P<0.05). Compared with group DHS,biochemical parameter of heart( BUN,SCr) of group DPre/Sev and group DPost/Sev were decreased(P<0.05). Compare with group DPre/Sev and group DPost/Sev,there was not significantly different(P>0.05).HE staining showed that group DS was no significant changed; swelling of renaltubular epitheli-al cells inthe. Group DHS, interstitial edema, Group DPre/Sev and group DPost/Sev were also decreased.3.4. BrainCompared with group DS,the S100β protein in brain tissue and plasma of group DHS, group DPre/Sev and group DPost/Sev were obviously increased(P<0.05). Compared with group DHS,the S100β protein in plasma and brain tissue of group DPre/Sev and group DPost/Sev decreased(P<0.05). HE staining showed that DS group is normal, DHS group was completely hurted, group DPre/Sev and group DPost/Sev group was less hurted than that of group DHS.3.5. LungCompared with group DS,the water content of lung of group DHS, group DPre/Sev and group DPost/Sev were increased(P<0.05). Compared with group DHS,the water content of lung of group DPre/Sev and group DPost/Sev were decreased(P<0.05). HE staining showed that group DS was normal, DHS group was completely hurted, group DPre/Sev and group DPost/Sev was less hurted than that of group DHS.3.6. IntestineCompared with the group DS, significantly increased plasma levels of I-FABP expression in the experimental group(P<0.05), compared with the group DHS, group DPre/Sev and DPost/Sev were obviously decreased(P<0.05). Compared with group DS, the water content of intestines of group DHS, group DPre/Sev and group DPost/Sev were increased(P<0.05).Compared with group DHS,the water content of intestines of group DPre/Sev and group DPost/Sev were decreased(P<0.05).Group DPre/Sev and group DPost/Sev could see slight enlargement, and little edematous. Compared with group DHS the hurt was relieved obviously.Conclusion:1.The heart, liver, kidney, brain,lung, intestine and other vital organs with hemorrhagic shock of Environmental hypothermia could be damaged.2. Sevoflurane Pre-conditioning or Post-conditioning on pigs with hemorrhagic shock of environmental hypothermia had organ protection.3. The protective effect of sevoflurane Pre-conditioning or Post-conditioning was associated with the suppression of inflammation,and increased the body,s antioxidant ability. |
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