Influence On The Concentration Of Plasma Endotoxin By Inhibition Of Complement Activation In Traumatic Hemorrhagic Shock Rats

Objectives:The changes of intestinal mucosa and plasma endotoxin bycomplement inhibition were observed in traumatic hemorrhagic shock rats usedcobra venom factor (CVF) , and we tried to identify probably an approach thatmight cause the change of LPS. We tried to investigate the influence of inhibitioncomplement on the level of serum tumor necrosis factor-α(TNF-α) and to evaluatethe inhibition of complement activation on the effect of remote organ pulmonaryinjury in rats secondary to traumatic hemorrhagic shock .Methods: Eighty of male SD rats were randomly divided into two groups:control and CVF treatment groups. In each group, the animals were killed at serialtime points: preshock and at 1, 6,and 24 hours postresuscitation, including ten ratsfor each time point. Twenty four hours before hemorrhage, rats were given anmainline dose of either 50 μg/kg CVF or an equal volume of saline solution.As severe trauma was generally incorporated with hemorrhagenic shock,liquid resuscitation was used in clinic treatment process. To imitate the clinicprocess, this experiment establishes an animal model of common clinic traumatichemorrhagic shock using rats with single fracture, parenchyma contusion andphlebotomy of one side-femoral. After animals were anesthetized and routinelysterilized the ventral skin, the celio were opened aseptically. The portal vein andinferior vena cava were seprated. The plasma and serum samples were collected todetect the concentration of LPS, the activity of CH50 and DAO, and the level ofTNF-αat various time points in two groups. Blood endotoxin concentrations weremeasured by the chromogenic Limulus Amebocyte Lysate (LAL) which wasmodified by perchloric acid (PCA) pretreatment for samples. Serum totalcomplement activation were detected by CH50 method, and small intestinaldiamine oxidase (DAO) were also measured by spectrometer. Blood TNF-αlevelswere detected by enzyme-linked immunosobernt assay (ELISA). Animals weresacrificed, tissued samples from lung and small intestine were harvested. Themorphologic changes of pulmonary and gut were observed with microscopy. Results: 1.Compared with preshock in control group, serum CH50 levels weredecreased promptly at 1 hour postresuscitation and restored gradually at 6 and 24hours phase . The levels of CH50 in the CVF group were always less than 5%during the experiment. 2.The concentration of plasma endotoxin in control group were increasedsignificantly and peaked at 1 hour after resuscitation. Endotoxin levels were comegradualy back to the basic level at 6 and 24 hours. Compared with the controlgroup, treatment with CVF could significantly reduced plasma endotoxin atvarious time points after resuscitation. But compared with preshock level,endotoxin concentrations at 1 hour were increased. The concentration of plasmaendotoxin between two groups had remarkable statistical difference at 1 and 6hours, and difference till 24 hour after resuscitation. 3.Serum DAO activities in control group were significantly higher at 1 hourthan preshock level, and were subsequently promptly decreased at 6 and 24 hours.Though serum DAO levels in CVF group also were increased, the current wasunconspicuous. Serum DAO levels between two groups had remarkable statisticaldifference at 1 hour point postresuscitation. 4.The levels of TNF-αin traumatic hemorrhagic shock rats were increasedsignificantly at 1hour point after resuscitation, and yet increased at 6 hours. Thepeak of TNF-αin CVF group was declined markedly at 1 hour postresuscitation,and was come rapidly back to the basic level at 6 and 24 hours points. The levelsof TNF-αbetween two groups had remarkable statistical difference at 1 and 6hours points. 5. With light microscope , we could observe that the tissues of gut wereinjured at 1 hour postresuscitation , severely damaged at 6 hour point andrecovered gradually at 24 hour phase. Compared with control group, CVF groupwas significantly decreased at serial time points after resuscitation. 6. Compared with preshock in control group, we could observe with lightmicroscope that remote organ pulmonary tissues were injured at 1 hourpostresuscitation, significantly damaged at 6 hour point and recovered at 24 hourphase. The morphologic changes of pulmonary tissue in CVF group were morelight than in control group at serial time points postresuscitation. 7. Serum CH50 in control group was negatively correlated with endotoxin,DAO ,TNF-αlevels as well as histologic injury scoring of intestinal tissue. SerumDAO activities were positively correlated with endotoxin, TNF-αlevels andhistologic injury scoring of intestine. In addition, significantly statisticalcorrelations existed between plasma endotoxin and serum TNF-αas well ashistologic injury scoring of intestinal tissue. However, serum CH50 in CVF groupwas not statistically correlated with endotoxin, DAO ,TNF-αlevels as well ashistologic injury scoring of intestinal tissue. Conclusions: 1.In traumatic hemorrhagic shock rats complement inhibitor could prevent theinjure of intestine and gut barrier dysfunction caused by complement activation,decrease endotoxin translocation and decline plasma endotoxin levels. 2.Combined with correlation analysis, it was suggested that complementactivation might play an important role in increasing the concentration of plasmaendotoxin through intestine injured in traumatic hemorrhagic shock rats.Complement inhibitor might prevent endotoxin translocation by intestinalprotection. 3.Inhibition of complement activation in traumatic hemorrhagic shock ratsmight decline the level of TNF-α. Combined with correlation analysis, complementactivation and the increasing of TNF-a levels might exist directly or indirectlythe interaction.4.Complement activation might cause remote organ pulmonary injury in ratssecondary to traumatic hemorrhagic shock . Complement inhibition mightsignificantly decrease lung injure.