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SUMMARY: Systemic inflammatory response syndrome (SIRS) is a potentially fatal reaction to various forms of tissue damage and infections that cause damage to various organs. Furthermore, the brain is damaged earlier than other organs, resulting in diffuse brain dysfunction. The central clinical symptom of SIRS is delirium and emotional changes are involved in disease development. Although the amygdala is known to play a major role, the mechanisms underlying emotional changes in the early stages of SIRS have not been elucidated. Therefore, changes to dopamine levels in the amygdala were observed using an in vivo model of lipopolysaccharide (LPS)- induced SIRS to clarify the biochemical mechanisms activated in the early stages of SIRS. Extracellular dopamine was collected from the amygdala of free moving rats via microdialysis and then analyzed by high-performance liquid chromatography. In addition, emotional changes were assessed with the open field and sucrose preference tests. In the LPS group, dopamine release in the amygdala increased remarkably immediately after LPS administration, peaking at 120 min. Thereafter, dopamine release temporarily decreased, but then significantly increased again after 180 min. The present results suggest that diffuse brain dysfunction in the early stages of SIRS may involve altered dopamine levels in the amygdala.
El síndrome de respuesta inflamatoria sistémica (SRIS) es una reacción potencialmente fatal a diversas formas de daño tisular e infecciones que causan injuria a varios órganos. Además, el cerebro se daña antes que otros órganos, lo que provoca una disfunción cerebral difusa. El síntoma clínico central del SIRS es el delirio y los cambios emocionales están involucrados en el desarrollo de la enfermedad. Aunque se sabe que la amígdala desempeña un papel importante, no se han dilucidado los mecanismos que subyacen a los cambios emocionales en las primeras etapas del SRIS. Por lo tanto, en el estudio se provocaron cambios en los niveles de dopamina en la amígdala utilizando un modelo in vivo de SRIS inducido por lipopolisacáridos (LPS) para dilucidar los mecanismos bioquímicos activados en las primeras etapas del SRIS. La dopamina extracelular se recogió de la amígdala de ratas en movimiento libre mediante microdiálisis y luego se analizó mediante cromatografía líquida de alta resolución. Además, se evaluaron los cambios emocionales con las pruebas de campo abierto y de preferencia de sacarosa. En el grupo de LPS, la liberación de dopamina en la amígdala aumentó de manera notable inmediatamente después de la administración de LPS, alcanzando un máximo a los 120 minutos. A partir de entonces, la liberación de dopamina disminuyó temporalmente, pero luego volvió a aumentar significativamente después de 180 min. Los resultadosactuales sugieren que la disfunción cerebral difusa en las primeras etapas del SIRS puede implicar niveles alterados de dopamina en la amígdala.
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Animals , Male , Rats , Dopamine , Systemic Inflammatory Response Syndrome , Amygdala , Lipopolysaccharides/toxicity , Cytokines , Rats, Sprague-Dawley , Systemic Inflammatory Response Syndrome/chemically inducedABSTRACT
Objective:To investigate the epidemiological characteristics of sepsis-associated encephalopathy (SAE) in patients with sepsis, analyze its risk factors and build a prediction model, which provides evidence for early clinical identification of SAE patients and improvement of clinical outcomes.Methods:A retrospective observational study was conducted. Sepsis patients admitted to the critical care medical center of the First Affiliated Hospital of Xinjiang Medical University from February 2022 to February 2023 were enrolled. According to whether SAE occurred, the patients were divided into sepsis group and SAE group. The 24 patients without sepsis in the same period were used as controls (non-sepsis group). Demographic data, relevant scores and laboratory test indicators at admission to intensive care unit (ICU), and prognostic indicators were collected. Univariate and multivariate Logistic regression analysis was used to analyze the risk factors for sepsis and SAE. Receiver operator characteristic curve (ROC curve) was drawn. The predictive value of each risk factor for sepsis and SAE.Results:A total of 130 patients with sepsis were included, of which 52 had SAE, and the incidence of SAE was 40.00%. There were significant differences in the length of ICU stay and total length of stay among all groups, while there were no significant differences in hospitalization cost and mechanical ventilation time. Multivariate Logistic regression analysis showed that pulmonary infection [odds ratio ( OR) = 46.817, 95% confidence interval (95% CI) was 5.624-389.757, P = 0.000], acute physiology and chronic health evaluation Ⅱ (APACHEⅡ: OR = 1.184, 95% CI was 1.032-1.358, P = 0.016), sequential organ failure assessment (SOFA: OR = 9.717, 95% CI was 2.618-36.068, P = 0.001), Charson comorbidity index (CCI: OR = 4.836, 95% CI was 1.860-12.577, P = 0.001), hemoglobin (Hb: OR = 0.893, 95% CI was 0.826-0.966, P = 0.005), glutamyltranspeptidase ( OR = 1.026, 95% CI was 1.008-1.045, P = 0.006) were independent risk factors for sepsis in ICU patients. Pulmonary infection ( OR = 28.795, 95% CI was 3.296-251.553, P = 0.002), APACHEⅡ score ( OR = 1.273, 95% CI was 1.104-1.467, P = 0.001), SOFA score ( OR = 8.670, 95% CI was 2.330-32.261, P = 0.001), CCI ( OR = 5.141, 95% CI was 1.961-13.475, P = 0.001), Hb ( OR = 0.922, 95% CI was 0.857-0.993, P = 0.031), glutamyltranspeptidase ( OR = 1.020, 95% CI was 1.002-1.038, P = 0.030) were independent risk factors for SAE in sepsis patients. ROC curve analysis showed that the area under the curve (AUC) of pulmonary infection, APACHEⅡ score, SOFA score, CCI, Hb, and glutamyltranspeptidase for predicting sepsis were 0.792, 0.728, 0.987, 0.933, 0.720, and 0.699, respectively; the AUC of the combined prediction of the above 6 variables for sepsis was 1.000, with a sensitivity of 100% and a specificity of 100%. The AUC predicted by pulmonary infection, APACHEⅡ score, SOFA score, CCI, and Hb for SAE were 0.776, 0.810, 0.907, 0.917, and 0.758, respectively; the AUC of the combined prediction of the above 5 variables for SAE was 0.975, with a sensitivity of 97.3% and a specificity of 93.1%. Conclusions:Sepsis is more severe when accompanied by encephalopathy. Pulmonary infection, Hb, APACHEⅡ score, SOFA score and CCI were independent risk factors of SAE. The combination of the above five indicators has good predictive value for early screening and prevention of the disease.
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Objective To explore the role of protective function of Sestrin2(Sesn2)to mitochondria in alleviating cognitive dysfunction in mice with sepsis-associated encephalopathy(SAE).Methods 6-week-old male C57BL/6J mice were randomly divided into three groups:sham group,CLP group and CLP plus eupatilin group,40 mice in each group.A sepsis model was induced by cecal ligation and perforation(CLP).The CLP plus eupatilin group was treated with eupatilin.Neurobehavioral test and Morris water maze(MWM)were used to deter-mine neurobehavior and spatial learning and memory function in mice.The number of neurons in hippocampal CA1 area was counted by Nissl staining.HT22 cells were randomly divided into a control group(Con),lipopolysaccha-ride group(LPS),LPS plus eupatilin treatment group(LPS plus eupatilin)and LPS plus eupatilin and Nrf2 siRNA treatment group(LPS plus eupatilin and si-Nrf2).Apoptosis was analyzed by terminal deoxynucleotidyl transferase-mediated nick end labeling(TUNEL)staining,Mitochondrial membrane potential(MMP)was used to analyze mitochondrial damage.Results Seven days after CLP,as compared with sham mice,Sesn2 in hippocampus and cortex decreased significantly in CLP mice(P<0.01).As compared with CLP group,the survival rate in CLP plus eupatilin group increased significantly(P<0.05).As compared with sham group,the mice in CLP group showed a relatively high nerve injury score(P<0.05),and had fewer platform crossings and shorter target stay time,while the mice in CLP plus eupatilin group exhibited a lower injury score(P<0.05),and stayed in the target area for a longer time(P<0.05).As compared with sham group,the co-localization rate of neurons,Sesn2 and Nrf2 in CLP group decreased significantly(P<0.05),and the number of CD68/Iba-1 positive microglia increased significantly(P<0.05),while CLP plus eupatilin group reversed these changes.As compared with Con group,apoptosis and MMP level in LPS group increased significantly(P<0.01),while apoptosis and MMP level in LPS plus eupatilin group were lower than those in LPS group(P<0.05).However,Nrf2 knockdown(LPS plus eupatilin and si-Nrf2 group)reversed the anti-apoptosis and mitochondrial protection of eupatilin.Conclusions Eupatilin can alleviate cognitive dysfunction and neurological deficit in SAE mice by activating Sesn2-Nrf2 pathway,and improve inflammatory microenvironment by alleviating mitochondrial dysfunction.
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@#BACKGROUND: Sepsis-associated encephalopathy (SAE) is a critical disease caused by sepsis. In addition to high mortality, SAE can also adversely affect life quality and lead to significant socioeconomic costs. This review aims to explore the development of evaluation animal models of SAE, giving insight into the direction of future research in terms of its pathophysiology and therapy. METHODS: We performed a literature search from January 1, 2000, to December 31, 2022, in MEDLINE, PubMed, EMBASE, and Web of Science using related keywords. Two independent researchers screened all the accessible articles based on the inclusion and exclusion criteria and collected the relevant data of the studies. RESULTS: The animal models for sepsis are commonly induced through cecal ligation and puncture (CLP) or lipopolysaccharide (LPS) injection. SAE can be evaluated using nervous reflex scores and sepsis evaluation during the acute phase, or through Morris water maze (MWM), open-field test, fear condition (FC) test, inhibitory avoidance, and other tests during the late phase. CONCLUSION: CLP and LPS injection are the most common methods for establishing SAE animal models. Nervous reflexs cores, MWM, FC test, and inhibitory avoidance are widely used in SAE model analysis. Future research should focus on establishing a standardized system for SAE development and analysis.
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Objective:To investigate whether stress hyperglycemia (SH) is an independent risk factor for the occurrence and mortality of sepsis-associated encephalopathy (SAE).Methods:From August 2016 to October 2021, sepsis patients admitted to the ICU of Guangdong Provincial People's Hospital were selected as the study subjects. According to whether they developed to SH (RBG>11.1 mmol/L) within 7 days of enrollment, the pat ients were divided into the SH group and the non-SH group for analysis. Logistic regression was used to analyze whether SH was an independent risk factor for SAE occurrence, and ROC curve was used to analyze the predictive value of SH to SAE. Kaplan-Meier curve was used to compare the 90-day survival of SAE patients with or without SH. Cox regression analysis was used to analyze the risk factors of 28-day and 90-day death in SAE patients.Results:A total of 183 sepsis patients were included, including 62 patients in the SH group and 121 in the non-SH group. Logistic regression analysis demonstrated that SH was an independent risk factor for SAE ( OR=4.452, 95% CI: 2.021-9.808, P <0.001). ROC curve demonstrated that SH could accurately predict SAE (AUC=0.831; Sensitivity=78.4%; Specificity=76.8%; and Yoden index=0.553). Kaplan-Meier curve demonstrated that the 90-day survival of SAE patients with SH significantly declined (log-rank test: P<0.01). Cox regression analysis suggested that SH was a risk factor for death at day 28 and day 90 in SAE patients (28 d, HR=2.272, 95% CI: 1.212-4.260, P=0.010; 90 d, HR=2.456, 95% CI: 1.400-4.306, P<0.01). Conclusions:SH is an independent risk factor for SAE and can predict SAE occurrence. SH significantly reduces 90-day survival and increase mortality at 28 and 90 days in SAE patients.
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Objective:To explore the clinical value of serum insulin combined with cardiac-related markers in evaluating the severity of sepsis associated encephalopathy (SAE).Methods:The clinical data of 130 children with sepsis who admitted to the Pediatric Intensive Care Unit of Hunan Children's Hospital from January 2018 to December 2021 were analyzed retrospectively, and the differences of serum insulin and cardiac-related markers in children with sepsis and SAE were compared.Results:The levels of serum insulin, creatine kinase isoenzyme, hypersensitive troponin T, and N-terminal cerebral urine peptide in the SAE group were significantly higher than those in the non-SAE group ( P<0.05), but there was no significant difference in heart rate and lactic acid ( P>0.05). The levels of serum insulin, creatine kinase isoenzyme, hypersensitive troponin T, N-terminal cerebral urine peptide and lactic acid in the death group were significantly higher than those in the survival group ( P<0.05), while the heart rate was not significantly different ( P>0.05). The area under ROC curve of serum insulin, creatine kinase isoenzyme, hypersensitive troponin T, and N-terminal cerebral urine peptide in predicting SAE were 0.841, 0.599, 0.700, and 0.667, respectively; in terms of judging the prognosis of sepsis, the area under ROC curve were 0.647, 0.669, 0.645, and 0.683, respectively; and in terms of judging the prognosis of children with SAE, the areas under the ROC curve were 0.509, 0.682, 0.666 and 0.555, respectively. Binary logistic regression equation was established with serum insulin, creatine kinase isoenzyme, hypersensitive troponin T, and N-terminal cerebral urine peptide: Y=8.153×NT-proBNP+1.704×CTnT-hs+27.121×insulin+0.946×CK-MB+1.573. The area under the ROC curve of the new variable Y in predicting sepsis SAE, evaluating the prognosis of sepsis, and predicting the prognosis of children with sepsis and SAE was 0.890, 0.756, and 0.729, respectively. Conclusions:Serum insulin, creatine kinase isoenzyme, hypersensitive troponin T, and N-terminal cerebral urine peptide can be used alone to determine the severity of sepsis and sepsis in children with SAE. The combined value of the four indicators is obviously better than that of the single indicator. The combined application of the four indicators may better evaluate the severity of sepsis and SAE.
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Sepsis-associated encephalopathy(SAE) is a diffuse brain dysfunction caused by sepsis.The main clinical manifestations are abnormal mental state, high mortality and poor prognosis.At present, there is no unified diagnostic standard for SAE.The exclusion diagnosis is mainly based on clinical symptoms and signs, combined with laboratory examination and imaging auxiliary examination.Among them, brain magnetic resonance imaging and quantitative electroencephalography can early detect brain dysfunction and predict the prognosis of children, which play an important role in the early diagnosis and prognosis assessment of SAE.Cerebral oxygen monitoring can dynamically reflect the changes of brain function and can be used for long-term monitoring of children with severe brain function injury.SAE is closely associated with poor prognosis, and mortality will increase with the extention of hospitalization.Therefore, early identification of SAE is of great significance to reduce mortality.
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Objective:To investigate the clinical diagnosis and prognosis in patients with emergency septic encephalopathy.Methods:Case data of 131 patients with septic encephalopathy admitted to the emergency department of Chuiyangliu Hospital Affiliated to Tsinghua University from January 2020 to December 2021 were selected and divided into survival group and death group. Logistic regression was used to analyze the risk factors affecting diagnosis, treatment and prognosis in patients with septic encephalopathy. Receiver operating characteristic (ROC) curve was used to evaluate the prognostic value of each indicator in patients with septic encephalopathy.Results:The mean arterial pressure (MAP) and pH level in the death group were lower than those in the survival group, while the C reactive protein (CRP), troponin T (TNI), D-dimer, lactic acid, creatinine, Acute Physiology and Chronic Health Evaluation Ⅱ (APACHE Ⅱ) score, Sequential Organ Failure Assessment (SOFA), proportion of ventilator support, proportion of vasoactive drug use in the death group were higher than those in the survival group, with statistically significant difference (all P<0.05). Multivariate logistic regression analysis showed that APACHEⅡ score ( OR=1.290, 95% CI: 1.121-1.485, P<0.001), SOFA score ( OR=1.447, 95% CI: 1.183-1.796, P<0.001), the proportion of vasoactive drug use ( OR=18.720, 95% CI: 4.486-78.108, P<0.001) could predict the prognosis of patients with septic encephalopathy, and the area under the curve (AUC) was 0.823, 0886, 0.787. Conclusions:Elderly age and underlying brain diseases are important factors in the occurrence of septic encephalopathy. APACHE Ⅱ score, SOFA score, and the proportion of vasoactive drug use can predict the prognosis of patients with septic encephalopathy.
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Objective:To evaluate the role of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) in sepsis-associated encephalopathy (SAE) and the relationship with pyroptosis in microglia of mice.Methods:Twenty-four SPF healthy male C57BL/6J mice, aged 6-8 weeks, weighing 18-22 g, were divided into 3 groups ( n=6 each) using a random number table method: sham operation group (Sham group), SAE group and SAE plus an NLRP3 inhibitor MCC950 group (SAE+ MCC950 group). The mouse model of SAE was prepared by cecal ligation and puncture after anesthesia. MCC950 20 mg/kg was intraperitoneally injected at 1 h after developing the model in SAE+ MCC950 group, and the equal volume of normal saline was given instead in the other groups. Open field tests were conducted at 1 day after developing the model to record the number of rearing and time spent in the central area. Novel object recognition tests were conducted at 2-3 days after developing the model to record the recognition index. After the behavioral experiment on 3 day after developing the model, mice were sacrificed and hippocampal tissues were collected for determination of the expression of NLRP3 (by Western blot), count of cells co-expressing NLRP3 and microglia-specific ionized calcium-binding adaptor molecule 1 (Iba-1) (by immunofluorescence), activity of caspase-1, and contents of interleukin-1beta(IL-1β) and IL-18 (by enzyme-linked immunosorbent assay). Results:Compared with Sham group, the number of rearing was significantly reduced, the time spent in the central area was shortened, the recognition index was decreased, the expression of NLRP3 was up-regulated, the count of NLRP3 + -Iba-1 + cells was increased, and the activity of caspase-1 and contents of IL-1β and IL-18 were increased in SAE and SAE+ MCC950 groups ( P<0.05). Compared with SAE group, the number of rearing was significantly increased, the time spent in the central area was prolonged, the recognition index was increased, the expression of NLRP3 was down-regulated, the count of NLRP3 + -Iba-1 + cells was decreased, and the activity of caspase-1 and contents of IL-1β and IL-18 were decreased in SAE+ MCC950 group ( P<0.05). Conclusions:NLRP3 is involved in the development of SAE, which may be related to the mediation in microglial pyroptosis in mice.
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Objective:To investigate the effect of electroacupuncture on calcium homeostasis in hippocampal neurons of mice with sepsis-associated encephalopathy (SAE).Methods:Twenty-four healthy male C57BL/6J mice, weighing 18-22 g, were divided into 4 groups ( n=6 each) using a random number table method: sham operation group (Sham group), SAE group, SAE plus electroacupuncture group (SAE+ EA group), and SAE plus sham electroacupuncture group (SAE+ SEA group). The virus carrying calcium ion (Ca 2+ ) fluorescent probes was injected and then an optical fiber was implanted into the hippocampal CA1 area to record the fluorescence signals of Ca 2+ . SAE was induced by cecal ligation and puncture in anesthetized mice at 3 weeks after administration. Starting from 3 days before surgery, Baihui and bilateral Quchi and bilateral Zusanli acupoints were stimulated for 30 min per day for 7 consecutive days in SAE+ EA group. In SAE+ SEA group, electroacupuncture was performed at the points 0.2 mm lateral to the corresponding acupoints without electrical stimulation. Open field tests were conducted at 5 days after surgery to record the number of rearing and changes in related Ca 2+ signals in hippocampal CA1 neurons. Novel object recognition tests were conducted at 6-7 days after surgery to record the recognition time and changes in related Ca 2+ signals in hippocampal CA1 neurons. Mice were sacrificed after the end of behavioral testing on 7 days after surgery, and brain tissues ipsilateral to the optical fiber implant were obtained and the fluorescence intensity of Ca 2+ in the hippocampal CA1 neurons was acquired using a fluorescent microscope. Results:Compared with Sham group, the number of rearing and amplitudes of related Ca 2+ signals in hippocampal CA1 neurons while rearing were significantly decreased in SAE group and SAE+ SEA group ( P<0.05), and no statistically significant changes were found in the parameters mentioned above in SAE+ EA group ( P>0.05), and the recognition index and amplitudes of related Ca 2+ signals while recognizing were significantly deceased, and the fluorescence intensity of Ca 2+ in hippocampal CA1 neurons was increased in SAE, SAE+ EA and SAE+ SEA groups ( P<0.05). Compared with SAE group and SAE+ SEA group, the number of rearing and amplitudes of related Ca 2+ signals in hippocampal CA1 neurons while rearing were significantly increased, the recognition index and amplitudes of related Ca 2+ signals in hippocampal CA1 neurons while recognizing were increased, and the fluorescence intensity of Ca 2+ in hippocampal CA1 neurons was decreased in SAE+ EA group ( P<0.05). There were no statistically significant differences in the parameters mentioned above between SAE group and SAE+ SEA group ( P>0.05). Conclusions:The mechanism by which electroacupuncture alleviates SAE may be related to regulation of Ca 2+ homeostasis in hippocampal neurons of mice.
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Objective:To evaluate the relationship between B-cell lymphoma/adenovirus E1B19 kDa-interacting protein 3-like protein (BNIP3L)/adenovirus E1B-interacting protein and mitochondrial dysfunction in the hippocampus of mice with sepsis-associated encephalopathy (SAE).Methods:One hundred and eighty C57BL/6J mice, aged 6-8 weeks, weighing 20-25 g, were divided into 4 groups ( n=45 each) using a random number table method: control group (C group), sham operation group (Sham group), SAE group, and SAE+ BNIP3L agonist carfilzomib group (SC group). The sepsis model was developed by cecal ligation and puncture (CLP) in anesthetized animals. In SC group, carfilzomib 2 mg/kg was intraperitoneally injected at 2 h after CLP. Twenty mice in each group were selected, and the survival at 7 days after operation was recorded. Eight surviving mice in each group were selected at 1 week after CLP for Morris water maze test. The remaining mice were sacrificed at 24 h after surgery, and the hippocampal tissues were harvested for determination of the expression of BNIP3L (by immunofluorescence) and BNIP3L in mitochondrial protein (by Western blot) and for microscopic examination of the morphological structure of mitochondria. The mitochondrial ATP content was measured by fluorescein-fluorescence enzyme luminescence method, and the mitochondrial membrane potential (MMP) was measured by fluorescence spectrophotometry. Results:Compared with C and Sham groups, the survival rate was significantly decreased, the escape latency was prolonged, the time of staying at the original platform quadrant was shortened, and the number of crossing the original platform region was decreased, the expression of BNIP3L in the hippocampal mitochondria was down-regulated, the MMP and content of mitochondrial ATP were decreased ( P<0.05), the intensity of fluorescence of BNIP3L in the hippocampus was decreased, and the damage to mitochondrial ultrastructure was marked in SAE group. Compared with SAE group, the survival rate was significantly increased, the escape latency was shortened, the time of staying at the original platform quadrant was prolonged, and the number of crossing the original platform region was increased, the expression of BNIP3L in the hippocampal mitochondria was up-regulated, the MMP and content of mitochondrial ATP were increased ( P<0.05), the intensity of fluorescence of BNIP3L in the hippocampus was decreased, and the damage to mitochondrial ultrastructure was attenuated in SC group. Conclusions:BNIP3L-mediated mitochondrial dysfunction may be involved in the mechanism of SAE developed in mice.
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Objective:To evaluate the role of autophagy in electroacupuncture (EA)-induced improvement in sepsis-associated encephalopathy (SAE) in mice.Methods:A total of 135 healthy adult male mice, aged 8-12 weeks, weighing 22-25 g, were used in this study. Ten mice were randomly selected to prepare caecal slurry after anesthesia. The remaining 125 mice were divided into 5 groups ( n=25 each) using a random number table method: sham operation group (group Sham), SAE group, SAE+ EA group (group EA), SEA+ EA+ autophagy agonist rapamycin group (group SAE+ EA+ R), and SAE+ EA+ autophagy inhibitor 3-methyladenine group (group SAE+ EA+ MA). SAE was induced by intraperitoneal injection of cecal slurry 200 μl. Bilateral Zusanli (ST36) acupoints were stimulated at 2, 24, 48 and 72 h after surgery in group SAE+ EA, group SAE+ EA+ R and group SAE+ EA+ MA. Autophagy agonist rapamycin 10 mg/kg and autophagy inhibitor 3-methyladenine 15 mg/kg were intraperitoneally injected at 30 min before EA in SAE+ EA+ R group and SAE+ EA+ MA group, respectively. The survival of mice was recorded at 7 days after developing the model. Ten mice were selected from each group at 8-12 days after developing the model, and the learning and memory ability was assessed by Morris water maze test. Five mice from each group were sacrificed after anesthesia, brains were removed, and hippocampal tissues were obtained for determination of contents of interleukin-1beta (IL-1β), IL-18 and tumor necrosis factor-α (TNF-α) (by enzyme-linked immunosorbent assay) and expression of p62, autophagy-related protein 16 like protein 1 (ATG16L1), and nucleotide like receptor protein 3 (NLRP3) (by Western blot). Results:Compared with Sham group, the survival rate at 7 days after developing the model was significantly decreased in the other 4 groups ( P<0.01). There was no significant difference in the survival rate at 7 days after developing the model among SAE group, SAE+ EA group, SAE+ EA+ R group and SAE+ EA+ MA group ( P>0.05). Compared with Sham group, the activity time at the target quadrant was significantly shortened, the escape latency was prolonged, the number of crossing the original platform was reduced, the contents of TNF-α, IL-1β and IL-18 were increased, the expression of ATG16L1 was down-regulated, and the expression of p62 and NLRP3 was up-regulated in SAE group ( P<0.05). Compared with SAE group, the escape latency was significantly shortened, the activity time at the target quadrant was prolonged, the number of crossing the original platform was increased, the contents of TNF-α, IL-1β and IL-18 were decreased, the expression of ATG16L1 was up-regulated, and the expression of p62 and NLRP3 was down-regulated in SAE+ EA group ( P<0.05). Compared with SAE+ EA group, no significant change was found in the parameters of Morris water maze test ( P>0.05), the contents of TNF-α, IL-1β and IL-18 were significantly decreased, the expression of ATG16L1 was up-regulated, and the expression of NLRP3 and P62 was down-regulated in SAE+ EA+ R group, and the expression of ATG16L1 was significantly down-regulated, and the expression of p62 and NLRP3 was up-regulated in SAE+ EA+ MA group ( P<0.05). Conclusions:The mechanism by which EA improves SAE is related to promotion of autophagy in hippocampal neurons, inhibition of NLRP3 inflammasome activation, and alleviation of neuroinflammatory responses in mice.
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Objective:To identify the differentially expressed long-chain non-coding RNA(lncRNA) and mRNA using ribonucleic acid sequencing(RNA-seq), and construct a competing endogenous RNA(ceRNA) regulatory network in mice with sepsis-associated encephalopathy.Methods:Ten clean-grade healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 2 groups( n=5 each) using a random number table method: sham operation group(group Sham) and sepsis group(group Sepsis). Sepsis was induced by cecal ligation and puncture(CLP) in group Sepsis, while group Sham only underwent laparotomy without CLP. Morris water maze test and contextual fear conditioning test were performed to detect the cognitive function on 1 day before CLP and 3 days after CLP. Three mice were randomly sacrificed in group Sham, and 3 mice with the worst results in the cognitive function test were sacrificed in group Sepsis. The hippocampal tissues were obtained for RNA-seq via the BGISEQ-500 platform, and the differentially expressed mRNA and lncRNA were identified. The differentially expressed mRNAs and lncRNAs were visualized and analyzed by Dr. Tom platform provided by Shenzhen BGI Technology Service Co., Ltd., and the ceRNA regulatory network was constructed using the online visualization tool Cytoscape software. Results:Compared with group Sham, the escape latency was significantly prolonged, and the percentage of time of staying at the target quadrants and percentage of time spent freezing were decreased in group Sepsis( P<0.05). A total of 62 differentially expressed lncRNAs were obtained from RNA-seq, of which the expression of 45 lncRNAs was up-regulated and the expression of 17 lncRNAs was down-regulated.There were 282 differentially expressed mRNAs identified from RNA-seq, of which the expression of 173 mRNAs was up-regulated, and the expression of 109 mRNAs was down-regulated.Gene Ontology enrichment analysis revealed that the differentially expressed mRNAs were involved in biological processes such as memory, learning or memory, inflammatory responses, regulation of aging-related behavioral decline, and regulation of synaptic plasticity. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis showed that differentially expressed mRNAs were enriched in IL-17 signaling pathway, TNF signaling pathway, NF-κB signaling pathway and etc. KDA analysis was performed on the differentially expressed mRNAs to identify the key driver genes, and the results showed that Ch25h, Il6ra, Lcn2, Sgk1, Nr4a3, Osm, Saa3, Ccl7, Sqle, Dhcr24 were the key SAE genes.A competing endogenous RNA regulatory network was successfully constructed based on 9 lncRNAs, 28 mRNAs and 134 miRNAs in the hippocampus of mice with SAE. Conclusions:The results of RNA-seq find that 10 mRNAs including Ch25h, Il6ra, Lcn2, Sgk1, Nr4a3, Osm, Saa3, Ccl7, Sqle, Dhcr24 and lncRNAs such as Rian, Gm35874 and Gm34347 are key genes regulating SAE in mice. Meanwhile, a ceRNA regulatory network based on lncRNA-miRNA-mRNA is successfully constructed in the hippocampus of mice with SAE.
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Sepsis-associated encephalopathy (SAE) is a diffuse brain dysfunction caused by sepsis, characterized by altered consciousness and cognitive dysfunction. Microglia, as the main immune cells in the brain, is an important factor in SAE progression. Microglia surface receptors play important roles in SAE pathogenesis by affecting microglia activation; in addition, microglia activation is involved in SAE by exacerbating neuroinflammation, impairing the blood-brain barrier and synaptic function. In this paper, the mechanism of microglial surface receptors and microglial activation in SAE development is reviewed to provide new ideas for SAE prevention and treatment.
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Objective:To evaluate the predictive value of regional cerebral oxygen saturation (rScO 2) for the occurrence of sepsis-associated encephalopathy (SAE). Methods:The data of 94 patients with sepsis admitted to the intensive care unit of Nanjing Drum Tower Hospital from September 2019 to June 2021 were collected. The patients were divided into SAE group and non-SAE group according to the evaluation results of daily intensive care unit confusion assessment method (CAM-ICU) during ICU treatment. The general data such as age and gender of the patients, rScO 2 on 1, 2, 3, 5, and 7 days of ICU admission, and prognostics were recorded. Receiver operator characteristic curve (ROC curve) was drawn to evaluate the predictive value of rScO 2 on SAE during ICU stay. Results:All 94 patients were enrolled in the analysis, of whom 59.6% (56/94) were male, and the mean age was (50.1±15.1) years old; the incidence of SAE was 31.9% (30/94). The levels of rScO 2 within first 3 days of ICU admission in the SAE group were significantly lower than those in the non-SAE group (1 day: 0.601±0.107 vs. 0.675±0.069, 2 days: 0.592±0.090 vs. 0.642±0.129, 3 days: 0.662±0.109 vs. 0.683±0.091, all P < 0.05). However, there was no significant difference in rScO 2 level on the 5th or the 7th day between the SAE and non-SAE groups (5 days: 0.636±0.065 vs. 0.662±0.080, 7 days: 0.662±0.088 vs. 0.690±0.077, both P > 0.05). ROC curve analysis showed that 1-day rScO 2 had the greatest predictive value for SAE [1 day: area under the ROC curve (AUC) = 0.77, 95% confidence interval (95% CI) was 0.65-0.89, P < 0.01; 2 days: AUC = 0.60, 95% CI was 0.48-0.72, P > 0.05; 3 days: AUC = 0.55, 95% CI was 0.41-0.68, P > 0.05]; with 1-day rScO 2 = 0.640 as the diagnostic threshold, the sensitivity was 73.4%, the specificity was 80.0%. Compared with the non-SAE group, the length of ICU stay and hospital stay in the SAE group were significantly longer [length of ICU stay (days): 13.6±7.1 vs. 9.0±4.3, length of hospital stay (days): 20.1±8.0 vs. 15.8±6.1, both P < 0.05], but the ICU mortality between the two groups was not statistically different. Conclusions:The incidence of SAE is relatively high in ICU patients, and the occurrence of SAE can be predicted by monitoring rScO 2. The rScO 2 value on the first day of ICU admission is closely related to the occurrence of SAE, and may be the target of sepsis resuscitation to guide the treatment and improve the long-term prognosis.
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Objective:To evaluate the effect of hydrogen-rich saline (HRS) on mitochondrial biogenesis and dynamics in hippocampus of mice with sepsis-associated encephalopathy (SAE).Methods:One hundred and twenty-eight male C57BL/6J mice, aged 6-8 weeks, weighing 20-25 g, were divided into 4 groups ( n=32 each) using a random number table method: sham operation group (Sham group), sham operation plus HRS group (Sham+ HRS group), SAE group and SAE plus HRS group.Sepsis was developed by cecal ligation and puncture (CLP) in anesthetized mice.HRS 10 ml/kg was intraperitoneally injected at 1 and 6 h after CLP in Sham+ HRS and SAE+ HRS groups.Twenty mice were randomly selected from each group to record the 7-day survival after operation.The working memory of the mice was observed by Y-maze test on days 3, 5 and 7 after CLP.The hippocampal tissues were obtained at 24 h after CLP for determination of the content of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and high-mobility group box 1 protein (HMGB1) (by enzyme-linked immunosorbent assay), activities of superoxide dismutase (SOD) and catalase (CAT) (by spectrophotometry), and expression of peroxisome proliferator-activated receptor gamma coactivator 1α (PGC-1α), nuclear respiratory factor 2 (NRF2), mitochondrial transcription factor A (Tfam), dynamin-related protein 1 (Drp1) and mitochondrial fusion protein mitofusin 2 (Mfn2) (by Western blot). Results:Compared with group Sham, the postoperative 7-day survival rate was significantly decreased, the time spent in novel arm was shortened, the contents of TNF-α, IL-6 and HMGB1 were increased, the activities of SOD and CAT were decreased, the expression of PGC-1α, NRF2 and Tfam was up-regulated, the expression of Drp1 was up-regulated, and the expression of Mfn2 was down-regulated in group SAE ( P<0.05). Compared with group SAE, the postoperative 7-day survival rate was significantly increased, the time spent in novel arm was prolonged, the contents of TNF-α, IL-6 and HMGB1 were decreased, the activities of SOD and CAT were increased, the expression of PGC-1α, NRF2 and Tfam was up-regulated, the expression of Drp1 was down-regulated, and the expression of Mfn2 was up-regulated in group SAE+ HRS ( P<0.05). Conclusions:The mechanism by which HRS alleviates SAE may be related to promotion of mitochondrial biogenesis, regulation of dynamics, and reduction of oxidative stress in hippocampus of mice.
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Objective:To explore the effect of pediatric critical illness score (PCIS), pediatric risk of mortality Ⅲ score (PRISM Ⅲ), pediatric logistic organ dysfunction 2 (PELOD-2), pediatric sequential organ failure assessment (p-SOFA) score and Glasglow coma scale (GCS) in the prognosis evaluation of septic-associated encephalopathy (SAE).Methods:The data of children with SAE admitted to the Pediatric Intensive Care Unit (PICU), Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences from January 2010 to December 2020 were retrospectively analyzed. They were divided into the survival and death groups according to the clinical outcome on the 28th day after admission. The efficiency of PCIS, PRISM Ⅲ, PELOD-2, p-SOFA and GCS scores for predicting death were evaluated by the area under the ROC curve (AUC). The Hosmer-Lemeshow goodness-of-fit test assessed the calibration of each scoring system.Results:Up to 28 d after admission, 72 of 82 children with SAE survived and 10 died, with a mortality rate of 12.20%. Compared with the survival group, the death group had significantly lower GCS [7 (3, 12) vs. 12 (8, 14)] and PCIS scores [76 (64, 82) vs. 82 (78, 88)], and significantly higher PRISM Ⅲ [14 (12, 17) vs. 7 (3, 12)], PELOD-2 [8 (5, 13) vs. 4 (2, 7)] and p-SOFA scores [11 (5, 12) vs. 6 (3, 9)] ( P<0.05). The AUCs of PCIS, PRISM Ⅲ, PELOD-2, p-SOFA and GCS scores for predicting SAE prognosis were 0.773 ( P=0.012, AUC>0.7), 0.832 ( P=0.02, AUC>0.7), 0.767 ( P=0.014, AUC>0.7), 0.688 ( P=0.084, AUC<0.7), and 0.692 ( P=0.077,AUC<0.7), respectively. Hosmer-Lemeshow goodness-of-fit test showed that PCIS ( χ2=5.329, P=0.722) predicted the mortality and the actual mortality in the best fitting effect, while PRISM Ⅲ ( χ2=12.877, P=0.177), PELOD-2 ( χ2=8.487, P=0.205), p-SOFA ( χ2=9.048, P=0.338) and GCS ( χ2=3.780, P=0.848) had poor fitting effect. Conclusions:The PCIS, PRISM Ⅲ and PELOD-2 scores have good predictive ability assessing the prognosis of children with SAE, while the PCIS score can more accurately evaluate the fitting effect of SAE prognosis prediction.
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@#BACKGROUND: Sepsis is a common cause of death in emergency departments and sepsis-associated encephalopathy (SAE) is a major complication. Rosuvastatin may play a neuroprotective role due to its protective effects on the vascular endothelium and its anti-inflammatory functions. Our study aimed to explore the potential protective function of rosuvastatin against SAE. METHODS: Sepsis patients without any neurological dysfunction on admission were prospectively enrolled in the “Rosuvastatin for Sepsis-Associated Acute Respiratory Distress Syndrome” study (SAILS trial, ClinicalTrials.gov number: NCT00979121). Patients were divided into rosuvastatin and placebo groups. This is a secondary analysis of the SAILS dataset. Baseline characteristics, therapy outcomes, and adverse drug events were compared between groups. RESULTS: A total of 86 patients were eligible for our study. Of these patients, 51 were treated with rosuvastatin. There were significantly fewer cases of SAE in the rosuvastatin group than in the placebo group (32.1% vs. 57.1%, P=0.028). However, creatine kinase levels were significantly higher in the rosuvastatin group than in the placebo group (233 [22-689] U/L vs. 79 [12-206] U/L, P=0.034). CONCLUSION: Rosuvastatin appears to have a protective role against SAE but may result in a higher incidence of adverse events.
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Objective:To evaluate the effect of high-concentration hydrogen inhalation on sepsis-associated encephalopathy (SAE) in mice.Methods:Healthy male ICR mice, aged 6-8 weeks, weighing 20-25 g, were divided into 4 groups ( n=50 each) using the random number table method: sham operation group (Sham group), SAE group, sham operation plus high-concentration hydrogen group (Sham+ H 2 group), and SAE plus high-concentration hydrogen group (SAE+ H 2 group). SAE model was prepared by cecal ligation and puncture (CLP) in anesthetized animals.At 1 and 6 h after operation, Sham+ H 2 and SAE+ H 2 groups inhaled the mixture of hydrogen and oxygen (67% hydrogen-33% oxygen) for 1 h, and Sham and SAE groups inhaled the mixture of nitrogen and oxygen (67% nitrogen-33% oxygen) for 1 h. The postoperative 7-day survival rate was recorded.Cognitive function was assessed by Y maze test at days 3, 5 and 7 after operation.The mice were sacrificed at 24 h after operation, and hippocampal tissues were obtained for microscopic examination of the pathological changes of neurons in hippocampal CA1 region (with a light microscope) and for determination of normal neuron count, contents of tumor necrosis factor-ɑ (TNF-α) and high mobility group box-1 (HMGB1) (by enzyme-linked immunosorbent assay), mitochondrial membrane potential (MMP) (by fluorescence spectrophotometry) and content of mitochondrial ATP (by fluorescein-fluorescent enzyme luminescence method). Results:Compared with Sham group, the 7-day survival rate after operation, percentage of spontaneous alternation at each time point after operation, and the number of normal neurons were significantly decreased, the contents of TNF-ɑ and HMGB1 were increased, and the contents of ATP and MMP were decreased in SAE and SAE+ H 2 groups ( P<0.05), and no significant change was found in Sham+ H 2 group ( P>0.05). Compared with SAE group, the 7-day survival rate after operation, percentage of spontaneous alternation at each time point after operation, and the number of normal neurons were significantly increased, the contents of TNF-ɑ and HMGB1 were decreased, and the contents of ATP and MMP were increased in SAE+ H 2 group ( P<0.05). Conclusions:High-concentration hydrogen inhalation can reduce SAE, and the mechanism may be related to reduction of hippocampal inflammatory responses and improvement in mitochondrial function in mice.
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Objective:To evaluate the role of mitophagy in cognitive dysfunction in rats with sepsis-associated encephalopathy (SAE).Methods:Twenty-four clean-grade healthy male Sprague-Dawley rats, aged 13-14 weeks, weighing 230-250 g, were divided into 3 groups ( n=8 each) using a random number table method: sham operation group (Sham group), SAE group and SAE+ autophagy inhibitor 3-methyladenine (3-MA) group (3-MA group).The SAE models were developed by cecal ligation and puncture in anesthetized animals.3-MA 10 mg/kg was intraperitoneally injected at 30 min after developing the model in 3-MA group.Cognitive function was assessed by Morris water maze test, and the escape latency and ratio of the time of staying at the target quadrant were recorded.After the end of Morris water maze test, the rats were sacrificed and hippocampal tissues were collected for microscopic examination of the pathological changes which were scored after hematoxylin-eosin staining and for determination of the expression of autophagy-related proteins LC3, Beclin1 and p62 (by Western blot).The ratio of LC3Ⅱ/LC3Ⅰwas calculated.The hippocampal mitochondria were isolated to measure mitochondrial membrane potential (MMP), ATP content and ATPase activity by spectrophotometry. Results:Compared with Sham group, the escape latency was significantly prolonged, the ratio of the time of staying at the target quadrant was decreased, the pathological score of hippocampus was decreased, and the contents of MMP and ATP and ATPase activity were decreased in SAE and 3-MA groups, the ratio of LC3Ⅱ/LC3Ⅰwas significantly increased, the expression of Beclin1 was up-regulated, and the expression of p62 was down-regulated in SAE group, and the ratio of LC3Ⅱ/LC3Ⅰwas significantly decreased, and the expression of Beclin1 and p62 was up-regulated in 3-MA group ( P<0.05).Compared with SAE group, the escape latency was significantly prolonged, the ratio of the time of staying at the target quadrant was decreased, the pathological score of hippocampus was decreased, the ratio of LC3/LC3Ⅰwas decreased, the expression of Beclin1 was down-regulated, the expression of p62 was up-regulated, and the contents of MMP and ATP and ATPase activity were decreased in 3-MA group ( P<0.05). Conclusions:Hippocampal mitophagy is involved in cognitive dysfunction in the rats with SAE.