Effects of Different Types of Early Restrictive Fluid Resuscitation on Immune Function and Multiorgan Damage on Hemorrhagic Shock Rat Model in a Hypothermic Environment

Abstract

Objective: This study was aimed at investigating the effects of different types of fluid restriction fluid resuscitation on the immune dysfunction and organ injury of hemorrhagic shock rats under a hypothermic environment.

Methods: SD rats were divided into sham operation group (SHAM), hemorrhagic shock model group (HS), crystal liquid limited resuscitation group (CRLLR), colloidal liquid limited resuscitation group (COLLR), and nonlimited resuscitation group (NLR); rats in each group were placed in a low-temperature environment of 0-5°C for 30 min, and then, a hemorrhagic shock rat model was prepared. Sodium lactate Ringer's restricted resuscitation solution, hydroxyethyl starch restricted resuscitation solution, and hydroxyethyl starch were used for resuscitation, and hemodynamic examination was performed. The mortality rate, inflammatory factors, oxidative stress factors, and immune function were detected by ELISA. The dysfunction and injury of the intestinal, lung, liver, and kidney were examined by histological methods.

Results: Hemorrhagic shock resulted in decreased immune function and activation of inflammation. Unrestricted fluid infusion further activated the inflammatory response. The crystalloid-restricted fluid infusion performed effectively to regulate inflammatory response, promote antioxidative activity, and reduce the immunosuppressive reaction. Rehydration could regulate the coagulation. The hydroxyethyl starch reduced the expression of platelet glycoproteins Ib and IIb/IIIa and blocked the binding of fibrinogen to activated platelets, thereby inhibiting intrinsic coagulation and platelet adhesion and aggregation. Rats in the CRLLR group showed to relieve the injury of the lung, liver, kidney, and intestine from hemorrhagic shock in low-temperature environment.

Conclusion: The early application of restrictive crystalloid resuscitation in hemorrhagic shock rats in hypothermic environment showed the best therapy results. Early LR-restrictive fluid replacement promotes the balance of inflammatory response and the recovery of immunosuppressive state, resists oxidative stress, stabilizes the balance of coagulation and fibrinolysis, improves coagulation function, and relieves organ injury.

Figure 1

Early restrictive fluid resuscitation regulates hemodynamic changes and coagulation function of HS rat model. (a) The percentage of survival rate of rats in different groups. (b) The times of mice heart rates in different groups. ns, P > 0.05; ∗, P < 0.05 vs. same group at T0; #, P < 0.05 vs. NLR at T2. (c) The MAP value of mice in different groups. ns, P > 0.05; ∗, P < 0.05 vs. same group at T0; $, P < 0.05 vs. COLLR at T2; #, P < 0.05 vs. NLR. SHAM: sham-operated group; HS: hemorrhagic shock model group; CRLLR: crystal liquid limited resuscitation group (CRLLR); COLLR: colloidal liquid limited resuscitation group; NLR: nonlimited resuscitation group.

Figure 2

The regulatory effect on coagulation function by early restrictive fluid resuscitation. (a) The R values of rats in different time points. (b) The K values of rats in different time points. (c) The α angle values of rats in different time points. (d) The MA values of rats in different time points. (e) The CI values of rats in different time points. (f) The LY30 values of rats in different time points; ns, P > 0.05; $, P < 0.05; #, P < 0.05 vs. COLLR at same time point; ∗, P < 0.05 vs. SHAM at same time point; SHAM: sham-operated group; HS: hemorrhagic shock model group; CRLLR: crystal liquid limited resuscitation group (CRLLR); COLLR: colloidal liquid limited resuscitation group; NLR: nonlimited resuscitation group.

Figure 3

The effect on inflammatory factors, oxidative stress factors, and immune function of rats under different fluid resuscitation treatment. (a) The concentration of TNF-α of rats in different groups. (b) The level of IL-6 of rats in different groups. (c) The concentration of IL-4 of rats in different groups. (d) The concentration of IL-10 of rats in different groups. (e) The concentration of MDA of rats in different groups. (f) The level of GSH of rats in different groups. ns, P > 0.05; $, P < 0.05; ∗, P < 0.05 vs. SHAM at same time point; #, P < 0.05 vs. COLLR at same time point. SHAM: sham-operated group; HS: hemorrhagic shock model group; CRLLR: crystal liquid limited resuscitation group (CRLLR); COLLR: colloidal liquid limited resuscitation group; NLR: nonlimited resuscitation group.

Figure 4

HS relieved the intestinal injury. (a) Western blot results of I-FABP. (b) The relative gray value of western blot results. (c) The ratio of wet to dry weight in rats' intestinal of different groups. ∗, P < 0.05 vs. SHAM; $, P < 0.05; #, P < 0.05 vs. COLLR. SHAM: sham-operated group; HS: hemorrhagic shock model group; CRLLR: crystal liquid limited resuscitation group (CRLLR); COLLR: colloidal liquid limited resuscitation group; NLR: nonlimited resuscitation group.

Figure 5

The lung injury resulted by HS treatment. (a) The ratio of dry to wet lung weight. ∗, P < 0.05 vs. SHAM; $, P < 0.05; #, P < 0.05 vs. COLLR. (b) The HE staining results of rats' lung tissue. Scale bar = 5 μm. SHAM: sham-operated group; HS: hemorrhagic shock model group; CRLLR: crystal liquid limited resuscitation group (CRLLR); COLLR: colloidal liquid limited resuscitation group; NLR: nonlimited resuscitation group.

Figure 6

The effect of liver and kidney dysfunction under HS treatment. (a) The levels of BUN and Cre in rat plasma of different groups. ∗, P < 0.05 vs. SHAM; $, P < 0.05; #, P < 0.05 vs. HS. (b) HE staining results in the rats' kidney. Scale bar = 5 μm. (c) The levels of ALT and AST in the rats' plasma. ∗, P < 0.05 vs. SHAM; $, P < 0.05; #, P < 0.05 vs. COLLR. SHAM: sham-operated group; HS: hemorrhagic shock model group; CRLLR: crystal liquid limited resuscitation group (CRLLR); COLLR: colloidal liquid limited resuscitation group; NLR: nonlimited resuscitation group.