Γ-secretase inhibitor DAPT prevents neuronal death and memory impairment in sepsis associated encephalopathy in septic rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Brain dysfunction is a frequent complication of sepsis, usually defined as sepsis-associated encephalopathy (SAE). Although the Notch signaling pathway has been proven to be involved in both ischemia and neuronal proliferation, its role in SAE is still unknown. Here, the effect of the Notch signaling pathway involved γ-secretase inhibitor DAPT on SAE in septic rats was investigated in a cecal ligation and puncture (CLP) model.</p><p><b>METHODS</b>Fifty-nine Sprague-Dawley rats were randomly divided into four groups, with the septic group receiving the CLP operation. Twenty-four hours after CLP or sham treatment, rats were sacrificed and their hippocampus was harvested for Western blot analysis. TNF-α expression was determined using an enzyme-linked immunosorbent assay (ELISA) kit. Neuronal apoptosis was assessed by TUNEL staining, and neuronal cell death was detected by H&E staining. Finally, a novel object recognition experiment was used to evaluate memory impairment.</p><p><b>RESULTS</b>Our data showed that sepsis can increase the expression of hippocampal Notch receptor intracellular domain (NICD) and poly (adenosine diphosphate [ADP]-ribose) polymerase-1 (PARP-1), as well as the inflammatory response, neuronal apoptosis, neuronal death, and memory dysfunction in rats. The γ-secretase inhibitor N-[N-(3,5-difluorophenacetyl)-1-alanyl]-S-phenylglycine t-butyl ester (DAPT) can significantly decrease the level of NICD and PARP-1, reduce hippocampal neuronal apoptosis and death, attenuate TNF-α release and rescue cognitive impairment caused by CLP.</p><p><b>CONCLUSION</b>The neuroprotective effect of DAPT on neuronal death and memory impairment in septic rats, which could be a new therapeutic approach for treating SAE in the future.</p>

The experimental study of ginkgolide B effectual on neuronal cell apoptosis in the rat with intracerebral hemorrhage / 中华急诊医学杂志

Objective To investigate the effects of ginkgolide B on neuronal cell apoptosis,superoxide dismutase activity,malondialdehyde,interleukin-1beta,tumor necrosis factor-alpha,and interleukin-6 levels in serum of rats with intracerebral hemorrhage in order to explore the role of ginkgolide B in suppressing the neuronal cell apoptosis.Methods A total of 175 male Wistar rats were randomly (random number)divided into sham operation group,intracerebral hemorrhage group,as well as low,medium and high dose treatment groups.The rat model of intracerebral hemorrhage was made with infusion of autologous whole blood to caudate nucleus in the right basal ganglia region.Ginkgolide B in dose of 5 mg/kg,10 mg/kg and 20 mg/kg was given to rats in the low,middle and high dose treatment groups by intraperitoneal injection once a day for 5 days after intracerebral hemorrhage.The rats with intracerebral hemorrhage in the sham operation groups received intraperitoneal administration of 1 mL saline.Animals were sacrificed by decapitation 2,6,12,24,48,72 h and 5 days after intracerebral hemorrhage.Brains were taken and blood samples were collected.Neuronal cell apoptosis was measured by using terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling(TUNEL),and superoxide dismutase activity in serum was determined by using xanthine oxidase method,and serum malondialdehyde level was detected by using thiobarbituric acid reactive substance assay,and interleukin-1beta,tumor necrosis factor-alpha,and interleukin-6 levels in serum were assayed with enzyme linked immunosorbent assay(ELISA).Statistical analysis was carried out by using one-way analysis of variance and least-significant difference test.Results As 2 h,6 h,12 h,24 h,48 h,72 h,and 5 days after intracerebral hemorrhage,the differences in the number of apoptotic neuronal cell,superoxide dismutase activity in serum,serum malondialdehyde,interleukin-1 beta,tumor necrosis factor-alpha and interleukin-6 levels between the low dose treatment group and intracerebral hemorrhage group were not significant statistically(P ＞0.05).As 12 h,24 h,72 h,and 5 days after intracerebral hemorrhage,the number of apoptotic neuronal cell,superoxide dismutase activity in serum,serum malondialdehyde,interleukin-1 beta,tumor necrosis factor-alpha and interleukin-6 levels in the medium dose and high dose treatment groups were significantly statistically lower than those in the intracerebral hemorrhage group(P ＜ 0.05),but these differences in above biomarkers were not significant statistically among these three groups 2 and 6 hours after intracerebral hemorrhage(P ＞ 0.05).Conclusions Ginkgolide B may lessen neuronal cell apoptosis by means of inhibition of free radical production and inflammatory reactions after intracerebral hemorrhage.

Change of microparticle procoagulant activity in patients with acute intracerebral hemorrhage / 中华急诊医学杂志

Objective To study the procoagulant activity of microparticles (MP) in patients with acute in-tracerebral hemorrhage (ICH) and to evaluate the correlation between procoagulant activity of MPs and disease out-come. Method From August 2006 through August 2008, 83 consecutive patients with history of hypertension ad-mitted for spontaneous basal ganglia hemorrhage including 54 male and 29 female, aged (60.9±9.7) years ranged from 41 to 79 years, were enrolled into this study. The control group was consisted of 30 age- and sex-matched (P= 0.429; P = 0.415) patients admitted for mild soft tissue injury. Patients with history of head trauma or previ-ous stroke, under the antiplatelet or anticoagulant medication, severe infection, or presence of previous cerebrovas-cttlar disease were excluded. Venous blood sample was kaken within the first 24 hours after disease onset. The MPs procoaulant potential was measured with a prothrombinase assay, and the levels of IL-6,TNF-α, D-dimer (DD)and thrombin-antithrombin Ⅲ complex (TAT) in plasma were measured with enzyme-linked immunosorbent assay. The multivariate analysis was made with forward stepwise logistic regression to determined the predictors of one. month mortality. The plasma levels of MPs were compared between ICH group and control group, between patients with intraventricular hemorrhage (IVH) and those without IVH,and between survivors and non-survivors with the Mann-Whitney U-test. The Spearman' s rank correlation coefficient was used to analyze the correlations between the plasma levels of MPs and ICH volume, Glasgow coma scale (GCS), and plasma levels of IL-6, TNF-α, DD and TAT. A receiver operating characteristic curve (ROC curve) identified the plasma MPs cutoff levels that predicted one-month mortality of patients. Under ROC curve, z statistic analysis was used to compare the area under curves (AUCs) between plasma IMPs and Glasgow coma scale, ICH volumes, and plasma levels of IL-6, TNF-α, DD and TAT for one-month mortality. Results Thirty-six patients (43.4%) died of ICH in a month. The multivariate analyses sorted out the GCS (odds ratio = 0.558, 95%CI:0.367-0.850, P = 0.007), Hematoma volume (odds ratio= 1.061, 95%C1:1.012- 1.113, P = 0.015) and IVH (odds ratio= 5.537, 95%CI:1.035-29.629, P = 0.045) as the independent pcedictors for one-week mortality. The MPs procoagulant activity in the ICH group (6.72±3.26 U/mL) was significantly higher than that in control group (1.84±0.82) U/mL (P = 0.000). The IMPs procoagulant activity in the non-survival group (8.51±3.45) U/mL was significantly higher than that in the survival group (5.35±2.33) U/mL (P = 0.000). The MPs procoagulant activity in the IVH group (7.66±3.39) U/mL was significantly higher than that in the non-lVH group (5.36±2.53) U/mL (P = 0.001). The MPs procoagulant activity was highly associated with GCS scores (r = -0.690, P = 0.000), ICH volumes (r =0.590, P = 0.000), and plasma IL-6 (r = 0.465, P = 0.015), TNF-α (r = 0.464, P = 0.016), DD(r= 0.567, P = 0.001) and TAT(r = 0.469, P = 0.014) in ICH. The ROC curve identified cutoff levels of MPs procoagulant activity to be 7.47 U/mL that predicted one-month mortality of patients with high sensitivity (77.8%) and specificity values (76.6%). Areas under curves (AUCs) of MPs procoagulant activity (AUC =0.825±0.048) were significantly larger than those of plasma IL-6 (AUC = 0.685±0.060, P = 0.042), TNF-α(AUC = 0.681±0.060, P =0.036) and TAT (AUC = 0.644±0.062, P =0.008).The AUCs ofMPs procoag-ulant activity were larger than those of plasma DD (AUC = 0.743±0.056), but this difference was not statistical significance (p = 0.226). Conclusions The procoagulant activity of MPs may contribute to the pathophysiology of ICH. The propcoagulant activity of MPs after spontaneous onset of ICH seems to correlate with clinical outcome in these patients. Its procoagulant activity can be used as an useful clinical marker for evaluating the prognosis of ICH.