Traumatic Hemorrhagic Shock Rats Research Articles

EFFECT OF MIR-21-3P ON INTESTINAL INJURY IN RATS WITH TRAUMATIC HEMORRHAGIC SHOCK RESUSCITATED WITH THE SODIUM BICARBONATE RINGER'S SOLUTION.

Background : This study aims to determine the impact and mechanism of miR-21-3p on intestinal injury and intestinal glycocalyx during fluid resuscitation in traumatic hemorrhagic shock (THS), and the different impacts of sodium lactate Ringer's solution (LRS) and sodium bicarbonate Ringer's solution (BRS) for resuscitation on intestinal damage. Methods : A rat model of THS was induced by hemorrhage from the left femur fracture. The pathological changes of intestinal tissues and glycocalyx structure were observed by hematoxylin-eosin staining and transmission electron microscope. MiR-21-3p expression in intestinal tissues was detected by real-time quantitative polymerase chain reaction. The expression of glycocalyx-, cell junction-, and PI3K/Akt/NF-κB signaling pathway-related proteins was analyzed by western blot. Results : MiR-21-3p expression was increased in THS rats, which was suppressed by resuscitation with BRS. BRS or LRS aggravated the intestinal injury and damaged intestinal glycocalyx in THS rats. The expression of SDC-1, HPA, β-catenin, MMP2, and MMP9 was upregulated, the expression of E-cad was downregulated, and the PI3K/Akt/NF-κB signaling pathway was activated in THS rats, which were further aggravated by BRS or LRS. The adverse effect of LRS was more serious than BRS. MiR-21-3p overexpression deteriorated the injury of intestinal tissues and intestinal glycocalyx; increased the expression of SDC-1, HPA, β-catenin, MMP2, and MMP9 while decreasing E-cad expression; and activated the PI3K/Akt/NF-κB signaling pathway in BRS-resuscitated THS rats. Conclusion : MiR-21-3p aggravated intestinal tissue injury and intestinal glycocalyx damage through activating PI3K/Akt/NF-κB signaling pathway in rats with THS resuscitated with BRS.

Effect of sodium bicarbonate Ringer's solution on lung injury in rats with traumatic hemorrhagic shock.

This study aimed to explore the impact of different pH values of resuscitation fluid on traumatic hemorrhagic shock (THS),focusing on their effects on glycocalyx and inflammation. A rat model of THS was induced by hemorrhage from a left femur fracture, while an oxygen-glucose deprivation/reoxygenation (OGD/R)-induced HULEC-5acell model was considered as an in vitro THSmodel. The lung tissue pathology and glycocalyx structure were assessed through hematoxylin-eosin (H&E) staining and transmission electron microscope examination. The levels of glycocalyx-related factors and inflammation-related factors were determined by enzyme-linked immunosorbent assay (ELISA).The expression of glycocalyx-related proteins, cell junction-related proteins, and proteins involved in the PI3K/Akt/NF-κB signaling pathway was analyzed by western blot. The results showed that both sodium bicarbonate Ringer's solution (BRS)and lactate Ringer's solution (LRS)were effective in restoring mean arterial pressure and heart rate in THS rats. However, LRS has a stronger impact on promoting inflammation and damaging the glycocalyx compared withBRS. In OGD/R-induced HULEC-5a cells, a pH of 7.4 and 6.5 increased inflammation and disrupted the glycocalyx, while a pH of 8.1 had no significant effect on inflammation or glycocalyx. Furthermore, the PI3K/Akt/NF-κBsignaling pathway was activated by fluid resuscitation and different pH values. However, the activating effect of BRS and pH 8.1 on the PI3K/Akt/NF-κBsignaling pathway was milder compared withLRS and pH6.5. In conclusion, an alkaline recovery environment was more beneficial for the treatment of THS.

MiR-148b caused liver injury in rats with traumatic hemorrhagic shock by inhibiting SIRT6 expression.

The purpose of this study was to investigate the role of miR-148b in liver injury in rats with traumatic hemorrhagic shock (THS) and to elucidate its potential mechanism. The levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the serum of rats were detected by enzyme-linked immune sorbent assay (ELISA), and the injury of rat liver was analyzed by hematoxylin-eosin (H&E) staining. Apoptosis of rat hepatocytes and normal rat liver cell line (BRL3A) was identified by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay and flow cytometry, respectively. MiR-148b and sirtuin 6 (SIRT6) expression was measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blot. Lactate dehydrogenase (LDH) content and cell viability were measured by commercial kits and cell counting kit-8 (CCK-8) assay, respectively. The binding sites of miR-148b and SIRT6 were predicted by the Starbase database and verified by dual luciferase reporter assay. MiR-148b expression in THS rats or ischemia-reperfusion (I/R)-treated cells was higher than in the control group. Overexpression of miR-148b further promoted the effects of I/R, which enhanced the levels of ALT, AST and LDH, cell apoptosis of liver tissue or BRL3A cells and decreased the expression of SITR6. Besides, miR-148b negatively correlated with SIRT6, and upregulated the expression of SIRT6 could partly reverse the effect of miR-148b. Hepatocyte injury induced by I/R was achieved by regulating miR-148b /SIRT6 axis.

Protective Effect of Crocin on Liver Function and Survival in Rats With Traumatic Hemorrhagic Shock

BackgroundThis study investigated the underlying mechanism of crocin in protecting rats with traumatic hemorrhagic shock (THS) from liver injury. Materials and MethodsEighty Sprague Dawley rats were randomly divided into four groups (n = 20), namely, Sham group, THS group, crocin group, and Sodium Acetate Ringer group. A rat model of THS was induced by hemorrhage from the left femur fracture. The effects of crocin on hemodynamics, cardiac output, blood gas, animal survival rate, and liver function in the rats with THS were determined, and its relationship with oxidative stress was also explored. ResultsCrocin significantly improved the survival rate, hemodynamic parameters, increased tissue blood flow, and promoted the liver function of the THS rats. Further results indicated that crocin significantly inhibited oxidative stress in serum and liver tissue of THS rats, with increased levels of superoxide dismutase, catalase, and glutathione, and also reduced levels of malondialdehyde and myeloperoxidase levels. In addition, crocin greatly increased nuclear factor erythroid 2-related factor 2/heme oxygenase-1 level in liver tissues of THS rats. ConclusionsThe protective mechanism of crocin on the liver of THS rats may be attributed to its abilities to stabilize hemodynamics, improve cardiac output and blood gas, increase antioxidant enzyme activity, reduce serum liver enzyme levels, and promote nuclear factor erythroid 2-related factor 2/heme oxygenase-1 pathway, thereby reducing oxidative stress.

Efficacy of Different Fluid Resuscitation Methods on Coagulation Function of Rats with Traumatic Hemorrhagic Shock

BackgroundFluid resuscitation is widely used for treating traumatic hemorrhagic shock. We focused on the efficacies of different fluid resuscitation methods on improving coagulation function of traumatic hemorrhagic shock (THS) rats. Materials and methodsSprague-Dawley rats (n = 100) were randomly divided into 5 groups, namely, Sham group, THS group, acetic acid Ringer's fluid (AR) group, hydroxyethyl starch solution (HES) group, and AR + HES group. A THS rat model was established by left femoral bleeding. The effects of different fluid resuscitation methods on conventional coagulation function parameters, Rotational thromboelastometry parameters, platelet-derived microparticles and endothelial cell–derived microparticles content of the THS rats were detected by ACL TOP system, rotation thromboelastometry, and flow cytometry, respectively. ResultsUsing AR and HES alone had no significant effect on the coagulation function of THS rats, but the two in combination reduced the increases of thrombin time, prothrombin time, activated part thrombin time, international normalized ratio, fibrin degradation products, D-dimer and the decreases of platelet count and fibrinogen concentration induced by THS. The CT and CFT were significantly reduced, whereas α and MCF were increased in the THS rats in AR + HES group. The combination of AR and HES reversed the effect of THS on elevating platelet-derived microparticles and endothelial cell–derived microparticle levels. In addition, the coagulation was relatively the optimal in the AR, HES, and AR + HES groups when the mice were resuscitated to a mean arterial pressure of 60 mmHg. ConclusionsAR combined with HES has a significant protective effect on coagulation function of THS rats when the mean arterial pressure reaches 60 mmHg.

The Calcilytic Drug Calhex-231 Ameliorates Vascular Hyporesponsiveness in Traumatic Hemorrhagic Shock by Inhibiting Oxidative Stress and miR-208a-Mediated Mitochondrial Fission

Background The calcium-sensing receptor (CaSR) plays a fundamental role in extracellular calcium homeostasis in humans. Surprisingly, CaSR is also expressed in nonhomeostatic tissues and is involved in regulating diverse cellular functions. The objective of this study was to determine if Calhex-231 (Cal), a negative modulator of CaSR, may be beneficial in the treatment of traumatic hemorrhagic shock (THS) by improving cardiovascular function and investigated the mechanisms. Methods Rats that had been subjected to THS and hypoxia-treated vascular smooth muscle cells (VSMCs) were used in this study. The effects of Cal on cardiovascular function, animal survival, hemodynamics, and vital organ function in THS rats and the relationship to oxidative stress, mitochondrial fusion-fission, and microRNA (miR-208a) were investigated. Results Cal significantly improved hemodynamics, elevated blood pressure, increased vital organ blood perfusion and local oxygen supply, and markedly improved the survival outcomes of THS rats. Furthermore, Cal significantly improved vascular reactivity after THS in vivo and in vitro. Cal also restored the THS-induced decrease in myosin light chain (MLC) phosphorylation (the key element for VSMC contraction). Inhibition of MLC phosphorylation antagonized the Cal-induced restoration of vascular reactivity following THS. Cal suppressed oxidative stress in THS rats and hypoxic-VSMCs. Meanwhile, THS induced expression of mitochondrial fission proteins Drp1 and Fis1 and decreased expression of mitochondrial fusion protein Mfn1 in vascular tissues. Cal reduced expression of Drp1 and Fis1. In hypoxic-VSMCs, Cal inhibited mitochondrial fragmentation and preserved mitochondrial morphology. In addition, miR-208a mimic decreased Fis1 expression, and miR-208a inhibitor prevented Cal-induced Fis1 downregulation in hypoxic-VSMCs. Conclusion Calhex-231 exhibits outstanding potential for effective therapy of traumatic hemorrhagic shock, and the beneficial effects result from its protection of vascular function via inhibition of oxidative stress and miR-208a-mediated mitochondrial fission.

Role of microRNA-15a-5p/TNFAIP3-interacting protein 2 axis in acute lung injury induced by traumatic hemorrhagic shock.

The present study aimed to investigate the role of microRNA (miR)-15a-5p in the pathogenesis of acute lung injury induced by traumatic hemorrhagic shock (THS), and to explore the underlying molecular mechanism. The expression level of miR-15a-5p was detected using reverse transcription-quantitative (RT-qPCR) and the association between miR-15a-5p and TNFAIP3-interacting protein 2 (TNIP2) was revealed using TargetScan and dual luciferase reporter assays. To investigate the effect of miR-15a-5p on THS-induced acute lung injury, a THS rat model was established. Lung capillary permeability and lung edema were then determined. Moreover, proinflammatory factors in the bronchoalveolar lavage fluid (BALF) and serum of the THS rat model were detected using ELISA. In addition, protein levels in the current study were measured via western blotting. It was revealed that miR-15a-5p was significantly upregulated in both patients with THS and samples from the THS rat model. TNIP2 represents a direct target of miR-15a-5p, and it was downregulated in both patients with THS and the THS rat model. Further analyses indicated that downregulation of miR-15a-5p significantly relieved acute lung injury induced by THS, evidenced by a decreased ratio of Evan's blue dye (EBD) in the BALF to EBD in plasma of THS rats, decreased lung permeability index and reduced lung wet/dry ratio. Inhibition of miR-15a-5p also decreased THS-induced upregulation of pro-inflammatory factors. Furthermore, the data revealed that THS-induced NF-κB activation in the lung tissues of rats was inhibited by miR-15a-5p knockdown. Moreover, it was demonstrated that all the effects of miR-15a-5p on THS rats were ablated following TNIP2 silencing. Taken together, the data of the current study indicate that miR-15a-5p downregulation serves a protective role in THS-induced acute lung injury via directly targeting TNIP2.

Protective effect of calcium-sensing receptor inhibitor Calhex231 on traumatic hemorrhagic shock rats

Objective To observe the protective effects of calcium-sensing receptor (CaSR) inhibitor Calhex231 on traumatic hemorrhagic shock rats. Methods 144 SD rats were divided into six groups by random number table method: normal group, shock group, lactated Ringer's solution (LR) group, LR+ Calhex231 0.1 mg/kg group, LR+ Calhex231 1 mg/kg group, and LR+ Calhex231 5 mg/kg group, with 16 rats in each group for survival observation and 8 rats for hemodynamics test. 64 SD rats were divided into four groups: normal group, shock group, lactated Ringer's solution (LR) group, LR + Calhex231 1 mg/kg group, with 8 rats in each group for detecting organ blood flow and superior mesenteric artery vascular reactivity and the other 8 rats for mesenteric artery vascular reactivity. After the establishment of traumatic hemorrhagic shock model, the shock group did not receive resuscitation, and the LR group was resuscitated with LR equal to two times of the blood loss volume. The three LR+ Calhex231 groups with different dosages were firstly given LR of equal volume to that of blood loss, and then the Calhex231 was dissolved into LR ( equal to the blood loss volume) to resuscitate. The wound was ligated and sutured immediately after resuscitation. The effect of Calhex231 on animal's 24-hour survival since the beginning of resuscitation was observed. The hemodynamics including the mean arterial blood pressure(MAP), left intraventricular systolic pressure(LVSP), maximal rising, and declining rate of left intraventricular pressure (±dp/dtmax) were observed before shock, at the end of shock, 1 hour after resuscitation, and 2 hours after resuscitation. The effects of Calhex231 on vital organ blood flow and vascular reactivity were observed 2 hours after resuscitation. Results All the shock rats died within 9 hours after the shock model was established. The survival outcomes of LR group rats were slightly improved compared with the shock group rats(P<0.05) . The survival time and 24 hour survival rate of LR+ Calhex231 1 mg/kg group and LR + Calhex231 5 mg/kg group rats were significantly increased compared with the shock group rats (P<0.05). The hemodynamic indexes of LR+ Calhex231 groups were higher than those of the LR group. The best effect was observed in LR + Calhex231 1 mg/kg group rats (P<0.01). The MAP, LVSP and ±dp/dtmax were restored to normal level (64.9%, 82.4%, 89.8%, and 77.8%, respectively). Meanwhile, the blood flow in liver and kidney of LR+ Calhex231 1 mg/kg group rats were increased from 57.2% and 41% to 108.7% and 95.1%, respectively. The vascular reactivity including superior mesenteric artery and mesenteric artery of LR+ Calhex231 1 mg/kg group rats were also increased (P<0.01). Conclusions In rats with hemorrhagic shock, the calcium sensitive receptor inhibitor Calhex231 can improve the vascular reactivity, the hemodynamics, and the blood flow of important organs. It plays a role in protecting the cardiovascular function and reducing the mortality after traumatic hemorrhagic shock. Key words: Receptors, calcium-sensing; Shock, hemorrhagic; Calhex231

4-Phenylbutyrate Benefits Traumatic Hemorrhagic Shock in Rats by Attenuating Oxidative Stress, Not by Attenuating Endoplasmic Reticulum Stress.

Vascular dysfunction such as vascular hyporeactivity following severe trauma and shock is a major cause of death in injured patients. Oxidative stress and endoplasmic reticulum stress play an important role in vascular dysfunction. The objective of the present study was to determine whether or not 4-phenylbutyrate can improve vascular dysfunction and elicit antishock effects by inhibiting oxidative and endoplasmic reticulum stress. Prospective, randomized, controlled laboratory experiment. State key laboratory of trauma, burns, and combined injury. Five hundred and fifty-two Sprague-Dawley rats. Rats were anesthetized, and a model of traumatic hemorrhagic shock was established by left femur fracture and hemorrhage. The effects of 4-phenylbutyrate (5, 20, 50, 100, 200, and 300 mg/kg) on vascular reactivity, animal survival, hemodynamics, and vital organ function in traumatic hemorrhagic shock rats and cultured vascular smooth muscle cells, and the relationship to oxidative stress and endoplasmic reticulum stress was observed. Lower doses of 4-phenylbutyrate significantly improved the vascular function, stabilized the hemodynamics, and increased the tissue blood flow and vital organ function in traumatic hemorrhagic shock rats, and markedly improved the survival outcomes. Among all dosages observed in the present study, 20 mg/kg of 4-phenylbutyrate had the best effect. Further results indicated that 4-phenylbutyrate significantly inhibited the oxidative stress, decreased shock-induced oxidative stress index such as the production of reactive oxygen species, increased the antioxidant enzyme levels such as superoxide dismutase, catalase, and glutathione, and improved the mitochondrial function by inhibiting the opening of the mitochondrial permeability transition pore in rat artery and vascular smooth muscle cells. In contrast, 4-phenylbutyrate did not affect the changes of endoplasmic reticulum stress markers following traumatic hemorrhagic shock. Furthermore, 4-phenylbutyrate increased the nuclear levels of nuclear factor-E2-related factor 2, and decreased the nuclear levels of nuclear factor κB in hypoxic vascular smooth muscle cells. 4-phenylbutyrate has beneficial effects for traumatic hemorrhagic shock including improving animal survival and protecting organ function. These beneficial effects of 4-phenylbutyrate in traumatic hemorrhagic shock result from its vascular function protection via attenuation of the oxidative stress and mitochondrial permeability transition pore opening. Nuclear factor-E2-related factor 2 and nuclear factor-κB may be involved in 4-phenylbutyrate-mediated inhibition of oxidative stress.

The changes of serum inflammatory cytokines and cyclooxygenase-2 mRNA in lung tissue and their role in secondary lung injury of traumatic hemorrhagic shock rats in dry heat enviroment of desert

Objective To study the changes of serum infflammatory cytokines cell interleukin-1 β (IL-1 β),interleukin-6 (IL-6) and cyclooxygenase-2 (COX-2) mRNA in lung tissues and their role in secondary lung injury of traumatic hemorrhagic shock rats in dry heat environment of desert.Methods 140 male SD rats were randomly divided into the dry heat environment and the normal temperature environment traumatic hemorrhagic shock group,respectively,each groups were randomly divided into 7 subgroup:the control group,after shock 0,0.5,1.0,1.5,2.0,3.0 h group,10 rats in each subgroup,After the establishment of traumatic hemorrhagic shock rat models in each environment,at the corresponding time points,serum and lung tissue were harvested.Pathological changes of lung tissue were observed,serum IL-1 β,IL-6 concentrations was detected with the method of enzyme-linked immunoassay (ELISA),and expression of COX-2 mRNA of lung tissue was detected by Real-time quantitative polymerase chain reaction.Results The dry heat envionment group has a higher lung histopathological injury score.The serum IL-1 β and expression of COX-2 mRNA (3.46 ± 0.19) in lung tissue in dry heat environment group were higher than that of the normal temperature environment group at each time point (2.13 ± 0.13),of which the peak appeared at 1.5 h in dry heat environment group,while the peak appeared at 2 h in the normal temperture environment group,the IL-6 peak appeared later than the IL-1 β in both environment groups.The concentration of serum IL-1 β,IL-6 and expression of COX-2 mRNA in lung tissue were positively correlated with pathological lung injury score in both environment groups (P ＜ 0.05).Conclusion The secondary lung injury of traumatic hemorrhagic shock was more serious and appead eralier in dry heat environment group than that of the normal temperature.The serum IL-1 β,IL-6 and the expression COX-2 may play and important role in the secondary lung injury of the traumatic hemorrhagic shock rats in the dry heat environment of the desert. Key words: Desert; Dry heat environment; Traumatic hemorrhagic shock ; Lung injury; Interleukin-1β ; Cell interleukin-6 ; Cyclooxygenase-2

Effects of lymph drainage on multiple organ injury of rats with traumatic hemorrhagic shock

Objective To observe the effect of shock lymph drainage on multiple organ injury of rats with traumatic hemorrhagic shock (THS) and discuss the relating mechanism. Methods Male Wistar rats were divided into control group, lymph drainage group and lymph return group. The THS model was established in lymph drainage group and lymph return group, when the shock mesenteric lymph was drained in lymph drainage group. The change of the mean arterial pressure ( MAP), the biochemical indices of liver, kidney, myocardium and acid-base, the morphology, ATP contents and ATPase activities of lung, kidney, liver and myocardium were observed. Results The MAP at multiple time points after 80 minutes of infusion, the ATP contents and ATPase activities of multiple organs in lymph drainage group were higher than those in lymph return group. Multiple biochemical indices in lymph drainage group were superior to those in lymph return group, with statistical difference. The inflammation, congestion, degeneration and necrosis were found in organs of lymph return group, but only mild lesions could be seen in lymph drainage group. Conclusions The shock lymph drainage can alleviate multiple organ injury of THS rats, mechanism of which is correlated with improvement of the energy metabolism and maintenance of MAP and acid-base status. Key words: Shock, hemorrhagic; Mesenteric lymph; Drainage; Emergy metabolism; Rats, Wistar