Carrimycin ameliorates lipopolysaccharide and cecal ligation and puncture-induced sepsis in mice

Carrimycin(CA),sanctioned by China’s National Medical Products Administration(NMPA)in 2019 for treating acute bronchitis and sinusitis,has recently been observed to exhibit multifaceted biological activities,encompassing anti-inflammatory,antiviral,and anti-tumor properties.Despite these applications,its efficacy in sepsis treatment remains unexplored.This study introduces a novel function of CA,demonstrating its capacity to mitigate sepsis induced by lipopolysaccharide(LPS)and cecal ligation and puncture(CLP)in mice models.Our research employed in vitro assays,real-time quantitative polymerase chain reaction(RT-qPCR),and RNA-seq analysis to establish that CA significantly reduces the levels of pro-inflammatory cytokines,namely tumor necrosis factor-alpha(TNF-α),interleukin 1 beta(IL-1β),and interleukin 6(IL-6),in response to LPS stimulation.Additionally,Western blotting and immunofluorescence assays revealed that CA impedes Nuclear Factor Kappa B(NF-κB)activation in LPS-stimulated RAW264.7 cells.Complementing these findings,in vivo experiments demonstrated that CA effectively alleviates LPS-and CLP-triggered organ inflammation in C57BL/6 mice.Further insights were gained through 16S sequencing,highlighting CA’s pivotal role in enhancing gut microbiota diversity and modulating metabolic pathways,particularly by augmenting the production of short-chain fatty acids in mice subjected to CLP.Notably,a comparative analysis revealed that CA’s anti-inflammatory efficacy surpasses that of equivalent doses of aspirin(ASP)and TIENAM.Collectively,these findings suggest that CA exhibits significant therapeutic potential in sepsis treatment.This discovery provides a foundational theoretical basis for the clinical application of CA in sepsis management.

Sodium tanshinone IIA sulfonate attenuates cardiac dysfunction and improves survival of rats with cecal ligation and puncture-induced sepsis

Cardiac dysfunction, a common consequence of sepsis, is the major contribution to morbidity and mortality in patients. Sodium tanshinone IIA sulfonate(STS) is a water-soluble derivative of Tanshinone IIA(TA), a main active component of Salvia miltiorrhiza Bunge, which has been widely used in China for the treatment of cardiovascular and cerebral system diseases. In the present study, the effect of STS on sepsis-induced cardiac dysfunction was investigated and its effect on survival rate of rats with sepsis was also evaluated. STS treatment could significantly decrease the serum levels of C-reactive protein(CRP), procalcitonin(PCT), cardiac troponin Ⅰ(cTn-Ⅰ), cardiac troponin T(cTn-T), and brain natriuretic peptide(BNP) in cecal ligation and puncture(CLP)-induced) septic rats and improve left ventricular function, particularly at 48 and 72 h after CLP. As the pathogenesis of septic myocardial dysfunction is attributable to dysregulated systemic inflammatory responses, several key cytokines, including tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), interleukin-6(IL-6), interleukin-10(IL-10) and high mobility group protein B1(HMGB1), were detected to reveal the possible mechanism of attenuation of septic myocardial dysfunction after being treated by STS. Our study showed that STS, especially at a high dose(15 mg×kg–1), could efficiently suppress inflammatory responses in myocardium and reduce myocardial necrosis through markedly reducing production of myocardial TNF-α, IL-6 and HMGB1. STS significantly improved the 18-day survival rate of rats with sepsis from 0% to 30%(P < 0.05). Therefore, STS could suppress inflammatory responses and improve left ventricular function in rats with sepsis, suggesting that it may be developed for the treatment of sepsis.

Accelerated Autophagy of Cecal Ligation and Puncture-Induced Myocardial Dysfunction and Its Correlation with Mammalian Target of Rapamycin Pathway in Rats

Background：Recent studies have indicated that autophagy is involved in sepsis-induced myocardial dysfunction.This study aimed to investigate the change of autophagy in cecal ligation and puncture （CLP）-induced myocardium dysfunction and its relationship with mammalian target of rapamycin （mTOR） pathway.Methods：Totally,12 rats were randomly divided into CLP group or sham-operated （SHAM） group.Cardiac tissues were harvested 18 h after CLP or sham operation.Pathology was detected by hematoxylin and eosin staining,cardiac functions by echocardiography,distribution ofmicrotubule-associated protein light chain 3 type Ⅱ （LC3II） by immunohistochemical staining,and autophagic vacuoles by transmission electron microscopy.Moreover,phosphorylation of mTOR （p-mTOR）,phosphorylation of S6 kinase-1 （PS6K1）,and LC3II and p62 expression were measured by western blotting.Pearson＇s correlation coefficient was used to analyze the correlation of two parameters.Results：The results by pathology and echocardiography revealed that there was obvious myocardial injury in CLP rats （left ventricle ejection fraction：SHAM 0.76 ± 0.06 vs.CLP 0.59 ± 0.l l,P 〈 0.01;fractional shortening：SHAM 0.51 ± 0.09 vs.CLP 0.37 ± 0.06,P 〈 0.05）.We also found that the autophagy process was elevated by CLE the ratio of LC3II/LC3I was increased （P 〈 0.05） while the expression of p62 was decreased （P 〈 0.05） in the CLP rats,and there were also more autophagosomes and autolysosomes in the CLP rats.Furthermore,the mTOR pathway in CLP myocardium was inhibited when compared with the sham-operated rats;p-mTOR （P 〈 0.01） and PS6K 1 （P 〈 0.05） were both significantly suppressed following CLP challenge.Interestingly,we found that the mTOR pathway was closely correlated with the autophagy processes.In our study,while p-mTOR in the myocardium was significantly correlated with p62 （r=0.66,P =0.02）,PS6K1 was significantly positively correlated with p62 （r =0.70,P =0.01） and negatively correlated with LC31I （r =-0.71,P =0.01）.Conclusions：The autophagy process in the myocardium was accelerated in CLP rats,which was closely correlated with the inhibition of the mTOR pathway.

Ligation of Caeca Improves Nitrogen Utilization and Decreases Urinary Uric Acid Excretion in Chickens

This study was carried out to clarify role of ceca in nitrogen nutrition of the chicken. Exp. 1： The effect of cecal ligation on nitrogen utilization and excretion was investigated in chickens fed 5% to 14% protein diet. Irrespective of dietary protein level and different protein sources, the ligation of cecal decreased uric acid excretion and tended to increase nitrogen utilization and balance with the exception of urea-added 10% protein diet. Exp. 2： The effect of cecal ligation on nitrogen utilization and excretion was investigated in conventional and colostomized chickens fed a 5% protein diet or 5% protein diet plus urea. Total nitrogen excretion and uric acid excretion increased by colostomy were depressed by cecal ligation in chickens. Therefore, nitrogen utilization and balance decreased by colostomy were increased by cecal ligation. Urinary nitrogen excretion was significantly decreased by cecal ligation in colostomized chickens, but the cecal ligation did not change fecal nitrogen excretion in chickens fed either diet. Exp. 3： This experiment was carried out to examine the effects of removal of cecal contents on nitrogen utilization, balance and nitrogen excretion in cecally ligated chickens. Total nitrogen excretion was significantly decreased by washing out the cecal contents with saline or antibiotics. The cecal ligation and the removal of cecal contents significantly decreased uric acid excretion in the excreta. There was a highly inverse relationship between microbial counts in the ceca and an excretory amount of uric acid. Exp. 4： In order to examine effects of cecal ligation on microbial activity, microbes were counted and products of microbial fermentation were determined. The ligation of caeca decreased microbial counts, concentrations of acetic, propionic and butyric acids and ammonia concentration of cecal contents. These results suggest that nitrogen metabolism in chickens is affected by possible changes in cecal fermentation caused by preventing the substances from urine and digesta from entering into the ceca.

Astragaloside IV Attenuates Polymicrobial Sepsis-Induced Cardiac Dysfunction in Rats via IKK/NF-κB Pathway

Objective To evaluate the protective effects of Astragaloside IV(AST)in a rat model of myocardial injury induced by cecal ligation and puncture(CLP).Methods The model of sepsis-induced cardiac dysfunction was induced by CLP.Using a random number table,50 specific pathogen free grade of Sprague Dawley rats were randomized into 5 groups:the sham group(sham),the model group(CLP,18 h/72 h)and AST group(18 h/72 h).Except the sham group,the rats in other groups received CLP surgery to induce sepsis.CLP groups received intragastric administration with normal saline after CLP.AST groups received intragastric administration with AST solution(40 mg/kg)once a day.The levels of inflammatory mediators and oxidative stress markers in the serum of the septic rats were determined via enzyme-linked immunosorbent assay(ELISA)at different time point,such as interleukin 6(IL-6),IL-10,high mobility group box-1 protein B1(HMGB-1),superoxide dismutase(SOD),and malondialdehyde(MDA).Cardiac function was determined by echocardiography.Moreover,changes in myocardial pathology were evaluated using hematoxylin and eosin staining.The levels of lactate dehydrogenase(LDH)and creatine kinase-MB(CK-MB)were analysed to determine the status of CLP-induced myocardium.In addition,the apotosis of myocardial cells was analysed by terminal-deoxynucleoitidyl transferase mediated nick end labeling(TUNEL).The protein levels of B-cell lymphoma-2(Bcl-2),Bcl-2-associated X(Bax),IκB kinaseα(IKKα),nuclear factor kappa B p65(NF-κB p65)were detected by Western blot analysis.Moreover,survival rate was investigated.Results AST improved the survival rate of CLP-induced rats by up to 33.3%(P<0.05).The cardioprotective effect of AST was observed by increased ejection fraction,fractional shortening and left ventricular internal diameter in diastole respectively(P<0.01 or P<0.05).Subsequently,AST attenuated CLP-induced myocardial apoptosis and the ratio of Bcl-2/Bax in the myocardium,as well as the histological alterations of myocardium(P<0.01 or P<0.05);the generation of inflammatory cytokines(IL-6,IL-10,HMGB-1)and oxidative stress markers(SOD,MDA)in the serum was significantly alleviated(P<0.01 or P<0.05).On the other hand,AST markedly suppressed CLP-induced accumulation of IKK-αand NF-κB p65 subunit phosphorylation(P<0.01 or P<0.05).Conclusions AST plays a significant protective role in sepsis-induced cardiac dysfunction and survival outcome.The possible mechanism of cardioprotection is dependent on the activation of the IKK/NF-κB pathway in cardiomyocytes.