Atorvastatin Improves Survival in Septic Rats: Effect on Tissue Inflammatory Pathway and on Insulin Signaling

The aim of the present study was to investigate whether the survival-improving effect of atorvastatin in sepsis is accompanied by a reduction in tissue activation of inflammatory pathways and, in parallel, an improvement in tissue insulin signaling in rats. Diffuse sepsis was induced by cecal ligation and puncture surgery (CLP) in male Wistar rats. Serum glucose and inflammatory cytokines levels were assessed 24 h after CLP. The effect of atorvastatin on survival of septic animals was investigated in parallel with insulin signaling and its modulators in liver, muscle and adipose tissue. Atorvastatin improves survival in septic rats and this improvement is accompanied by a marked improvement in insulin sensitivity, characterized by an increase in glucose disappearance rate during the insulin tolerance test. Sepsis induced an increase in the expression/activation of TLR4 and its downstream signaling JNK and IKK/NF-κB activation, and blunted insulin-induced insulin signaling in liver, muscle and adipose tissue; atorvastatin reversed all these alterations in parallel with a decrease in circulating levels of TNF-α and IL-6. In summary, this study demonstrates that atorvastatin treatment increased survival, with a significant effect upon insulin sensitivity, improving insulin signaling in peripheral tissues of rats during peritoneal-induced sepsis. The effect of atorvastatin on the suppression of the TLR-dependent inflammatory pathway may play a central role in regulation of insulin signaling and survival in sepsis insult

Esophagogastric junction adenocarcinoma: multivariate analyses of surgical morbi-mortality and adjuvant therapy

BACKGROUND: In recent years the literature has recorded a progressive increase in the prevalence of adenocarcinoma of the esophagogastric junction. Several factors can interfere with the morbidity and mortality of surgical treatment. AIM: Non-randomized retrospective study of prognostic factors of operated patients by adenocarcinoma of esophagogastric junction, with or without post-operative chemotherapy and radiotherapy. METHODS: Medical records were reviewed from patients treated at university hospital in the period of 1989 and 2009, to obtain data about pre and postoperative treatment. Cox's univariate and multivariate regression analysis of risk factors for prognostic of these patients were done with level of significance of 5 %. RESULTS: Were reviewed 103 patients distributed as: 1) 78 (75.7%) patients without adjuvant therapy, and 2) 25 (24.3%) with it. All patients underwent surgical resection with curative intent. Cox's multivariate regression analysis of all patients showed that: lymphnode invasion N2 had greater risk of death in 5.9 times; broncopneumonia, in 11.4 times; tumoral recurrence during clinical following greater in 3.8 times. CONCLUSION: Tumoral recurrence, lymphnode metastasis and broncopneumonia in the postoperative period were factors of bad prognosis and contributed significantly to increase morbimortality and decrease global survival.RACIONAL: Nos últimos anos a literatura tem registrado aumento progressivo da prevalência do adenocarcinoma da transição esofagogástrica. Vários fatores podem interferir na morbimortalidade do tratamento cirúrgico. OBJETIVO: Estudo retrospectivo não-randomizado dos fatores prognósticos dos pacientes operados por adenocarcinoma da transição esofagogástrica, com ou sem quimio e radioterapia pós-operatórias. MÉTODOS: Foram revistos os prontuários dos pacientes tratados em hospital universitário no período de 1989 a 2009, para obtenção de informações referente ao pré e pós-operatório. Análises de regressão univariada e multivariada de Cox dos fatores de risco para o prognóstico destes pacientes foram realizadas com nível de significância de 5 %. RESULTADOS: Foram incluídos 103 pacientes assim distribuídos: 1) 78 (75,7%) não submetidos ao tratamento adjuvante, e 2) 25 (24,3%) submetidos a ele. Todos os pacientes foram operados com intenção curativa (esofagectomia e/ou gastrectomia). A análise multivariada de toda a casuística mostrou a influência dos seguintes fatores na sobrevida: invasão linfonodal, pacientes com N2 tiveram risco de óbito 3,4 vezes maior que os com N0; com N3, 5,9 vezes maior; com broncopneumonia, 11,4 vezes maior; com recidiva tumoral durante o seguimento clínico 3,8 vezes maior. CONCLUSÃO: A recidiva tumoral, metástase linfonodal e broncopneumonia no pós-operatório foram fatores de piora no prognóstico, contribuindo significativamente para elevar a morbimortalidade e diminuindo a sobrevida global.22923

Representative blots show the JNK phosphorylation in liver (A), muscle (B) and adipose tissue (C) of sham and septic rats (upper panels).

<p>Total protein expression of JNK (A–C, lower panels). Phosphorylation of c-jun in liver (D), muscle (E) and adipose tissue (F) of sham and septic rats. Serine 307 Phosphorylation of IRS1 in liver (G), muscle (H) and adipose tissue (I) of sham and septic rats (upper panels). Total protein expression of IRS-1 (G–I, lower panels). Data are presented as means ± S.E.M from 6–8 rats per group. *P<0.05 (Sepsis/Sal vs. all others groups); **P<0.001 (Sepsis/Sal vs. control); #P<0.05 (Sepsis/Sal vs. Sepsis/Ator). IB, immunoblot; CLT: Sham/Saline; ShT: Sham/Atorvastatin; SAL: saline; ATOR: atorvastatin.</p

To evaluate the association of TLR4 with MyD88, immunoprecipitations were performed with MyD88 antibody followed by immunoblotting with TLR4 specific antibody.

<p>Representative blots show TLR4 activation (upper panels) and expression (lower panels) in liver (A), muscle (B) and adipose tissue (C) of sham and septic rats. IKKβ phosphorylation in liver (D), muscle (E) and adipose (F) of sham and septic rats. Total protein expression of IKKβ (D–F, lower panels). Phosphorylation of IκBα in liver (G), muscle (H) and adipose (I) of sham and septic rats. Data are presented as means ±S.E.M from 6–8 rats per group. *P<0.05 (Sepsis/Sal vs. all other groups); **P<0.001 (Sepsis/Sal vs. control); #P<0.05 (Sepsis/Sal vs. Sepsis/Ator). IB, immunoblot; CLT: Sham/Saline; ShT: Sham/Atorvastatin; SAL: saline; ATOR: atorvastatin.</p

Effects of atorvastatin treatment on insulin signaling in the CLP rat.

<p>Representative blots show insulin-induced tyrosine phosphorylation of Insulin Receptor β (IRβ) in liver (A), muscle (B) and adipose (C) of sham and septic rats. Total protein expression of IRβ (A–C, lower panels). Insulin-induced tyrosine phosphorylation of Insulin Receptor Substrate 1 (IRS1) in liver (D), muscle (E) and adipose tissue (F) of sham and septic rats. Total protein expression of IRS1 (D–F, lower panels). Insulin-induced serine phosphorylation of Akt in liver (G), muscle (H) and adipose (I) of sham and septic rats. Insulin-induced threonine phosphorylation and total protein expression of Akt (G–I, lower panels). In this case, blots were stripped and reprobed with β-actin (A–I, lower panels) to confirm equal loading of proteins. Data are presented as means +/− S.E.M from 6–8 rats per group. *P<0.05 (Sepsis/Sal vs. all others groups). IB, immunoblot; CLT: Sham/Saline; ShT: Sham/Atorvastatin; SAL: saline; ATOR: atorvastatin.</p

Representative blots show the NFkB activation in nuclear fractions of liver (A), muscle (B) and adipose tissue (C) of sham and septic rats.

<p>In this case blots were stripped and reprobed with actin (A–C, lower panels) to confirm equal loading of proteins. Tissue levels of iNOS (D–F) and IL-6 (G–I) expression in liver, muscle and adipose tissue of sham and septic rats. Data are presented as means ± S.E.M from 6–8 rats per group. *P<0.05 (Sepsis/Sal vs. all others groups); **P<0.001 (Sepsis/Sal vs. control); #P<0.05 (Sepsis/Sal vs. Sepsis/Ator). IB, immunoblot; CLT: Sham/Saline; ShT: Sham/Atorvastatin; SAL: saline; ATOR: atorvastatin.</p

Effect of atorvastatin on survival in CLP sepsis model.

<p>Male Wistars rats, 8 weeks old, were given saline (Sepsis/Sal, n = 20) or atorvastatin 10 mg/kg (Sepsis/Ator, n = 20), 3 h and once a day after CLP. Survival of the rats was monitored at intervals of 12 h for 15 days. The overall difference in survival rate between the groups with and without atorvastatin was significant (P<0.0001) (A). Fasting blood glucose (B). Fasting insulin levels (C). Glucose disappearance rate (D). HOMA-IR index (E). Serum levels of TNF-α (F) and IL-6 (G). Data are presented as means and S.E. of six to eight rats per group. *P<0.05 (Sepsis saline vs. all others groups).</p