Effects of Qingwen Baidu decoction on coagulation and multiple organ injury in rat models of sepsis

Background:Sepsis-induced coagulopathy and multiple organ dysfunction syndromes are the leading causes of death in patients with sepsis.Qingwen Baidu decoction(QWBD)can effectively improve the clinical manifestations of sepsis and ease inflammation,but its effects on coagulation functions and multiple organ injuries remain unclear.Methods:100 healthy,male Sprague-Dawley rats were randomly divided into the sham group,the cecal ligation and puncture(CLP)group,the low-dose QWBD group,and the high-dose QWBD group,with 25 rats in each group.The sepsis model was established using CLP.Blood was collected to measure platelet count,serum creatinine(Cr),blood urea nitrogen(BUN),alanine aminotransferase(ALT),and aspartate aminotransferase(AST)levels,as well as coagulation function.The total protein in bronchoalveolar lavage fluid(BALF)was determined in each group of rats.The lung,liver,and kidney tissues were harvested,and statistics were calculated on the wet-to-dry(W/D)weight ratio.Changes in histopathology and thrombin level were evaluated in each group.The remaining ten rats in each group were observed daily to record the number of surviving rats.Such observation was made consecutively for 7 days to calculate survival rates.Results:After model establishment,ALT,AST,Cr,and BUN levels were significantly elevated(P<0.01).The BALF protein content and lung W/D weight ratio were significantly increased(P<0.01).Furthermore,the survival rate of rats was significantly reduced in the CLP group compared with the sham group.After the treatment,rats in the high-dose QWBD group had lower ALT(P<0.05),AST(P<0.01),Cr(P<0.05),BUN(P<0.01)levels,lower BALF protein content(P<0.05)and lower lung W/D weight ratio(P<0.01)than the CLP group.However,rats in the high-dose QWBD group had significantly better pathological changes in the lung,liver,and kidney compared to the sham group.After the treatment,the platelet level in the peripheral blood was elevated(P<0.05)and both activated partial thromboplastin time and prothrombin time were significantly shortened(P<0.01).The fibrinogen level was significantly increased(P<0.01).Finally,thrombin positive expression areas in the lung,liver,and kidney were significantly decreased in the high-dose QWBD group.Conclusion:QWBD can improve coagulation disorders caused by sepsis and has a protective effect on multiple organ injuries in rats.

edox factor-1 may mediate the repair of multiple organ injuries lfter fiver transplantation

Background Apurinic apyrimidinic endonuclease/redox effector factor 1 （APE1/Ref-1） is an important enzyme in the repair of reactive oxygen species-induced DNA damage, and its expression/activation can be induced by reactive oxygen species. The aim of this research was to investigate the relationship between multiple-organ injuries and expression of Ref-1 in the early period after liver transplantation. Methods One hundred and fifty adult male Wistar rats were divided randomly into three groups： liver transplantation, sham surgery, and untreated control. After liver transplantation, animals were sacrificed at different time points. Hepatic and renal functions were analyzed by serology. Histology, apoptotic levels, and Ref-1 expression were examined by immunohistochemistry in the liver, kidneys, intestines, and lungs. Results Serum levels of alanine aminotransferase and aspartate aminotransferase peaked 6 hours after liver transplantation and decreased appreciably after 12 hours in the transplantatior~ group, suggesting that the degree of liver injury in the early period after transplantation peaked at 6 hours and then decreased. Pathological analyses showed that hepatic tissues were more severely injured in the transplantation group than in the sham and untreated groups. A considerable number of infiltrating inflammatory cells was observed around the portal vein in the transplantation group. Injuries to the kidneys, intestines, and lungs were milder after liver transplantation. Apoptotic levels increased after liver transplantation in all four organs examined. Ref-1 expression was higher in the transplantation group in the early period after liver transplantation than in the sham surgery and untreated control groups. Conclusion Ref-1 expression induced by ischemia-reperfusion injury may have a critical role in repairing multiple-organ injuries after liver transplantation.

Protective Effects of Erythropoietin on Endotoxin-related Organ Injury in Rats

Summary： The protective effect of erythropoietin （EPO） on tissues following ischemia and reperfusion injuries remains poorly understood. We aimed to investigate the effect of EPO in preventing en- dotoxin-induced organ damage. Rat model of multiple organ failure （MOF） was established by tail vein injection of 10 mg/kg lipopolysaccharide （LPS）. Recombinant human EPO treatment （5000 U/kg） was administered by tail vein injection at 30 min after LPS challenge. Twenty-four h after EPO treatment, changes in serum enzyme levels, including aspartate aminotransferase （AST）, alanine transaminase （ALT）, blood urea nitrogen （BUN） and creatinine （Cr）, were evaluated by biochemical analysis. Serum levels of tumor necrosis factor-tx （TNF-ct） were determined by using immunoradiometric assay. Histological examination of tissue sections was carried out by hematoxylin and eosin staining, while ul- trastructure evaluation of organ tissues was assessed by transmission electron microscopy. Protein ex- pression levels were detected by using Western blotting. EPO treatment showed a modest effect in pre- venting LPS-induced elevation of AST, ALT, BUN, Cr, and TNF-ct levels, and in protecting against LPS-induced tissue degeneration and injured ultrastructure in the lung, liver, and kidney. Moreover, LPS promoted phosphorylation of alanine aminotransferase （AKT） and increased nuclear factor-r,B （NF-rB） activation in the lung, liver, and kidney （P〈0.05 vs. control）. However, EPO treatment significantly de- creased the LPS-induced pAKT up-regulation in these tissues （P〈0.05 vs. LPS treatment alone）. The present study demonstrates that EPO may play a protective role against LPS-induced MOF by reducing the inflammatory response and tissue degeneration, possibly via the phosphatidylinositol 3-kinase/AKT and NF-r,B signaling pathways.

Intratracheal administration of umbilical cord-derived mesenchymal stem cells attenuates hyperoxia-induced multi-organ injury via heme oxygenase-1 and JAK/STAT pathways

BACKGROUND Bronchopulmonary dysplasia(BPD)is not merely a chronic lung disease,but a systemic condition with multiple organs implications predominantly associated with hyperoxia exposure.Despite advances in current management strategies,limited progress has been made in reducing the BPD-related systemic damage.Meanwhile,although the protective effects of human umbilical cord-derived mesenchymal stem cells(hUC-MSCs)or their exosomes on hyperoxia-induced lung injury have been explored by many researchers,the underlying mechanism has not been addressed in detail,and few studies have focused on the therapeutic effect on systemic multiple organ injury.AIM To investigate whether hUC-MSC intratracheal administration could attenuate hyperoxia-induced lung,heart,and kidney injuries and the underlying regulatory mechanisms.METHODS Neonatal rats were exposed to hyperoxia(80%O_(2)),treated with hUC-MSCs intratracheal(iT)or intraperitoneal(iP)on postnatal day 7,and harvested on postnatal day 21.The tissue sections of the lung,heart,and kidney were analyzed morphometrically.Protein contents of the bronchoalveolar lavage fluid(BALF),myeloper oxidase(MPO)expression,and malondialdehyde(MDA)levels were examined.Pulmonary inflammatory cytokines were measured via enzyme-linked immunosorbent assay.A comparative transcriptomic analysis of differentially expressed genes(DEGs)in lung tissue was conducted via RNA-sequencing.Subsequently,we performed reverse transcription-quantitative polymerase chain reaction and western blot analysis to explore the expression of target mRNA and proteins related to inflammatory and oxidative responses.RESULTS iT hUC-MSCs administration improved pulmonary alveolarization and angiogenesis(P<0.01,P<0.01,P<0.001,and P<0.05 for mean linear intercept,septal counts,vascular medial thickness index,and microvessel density respectively).Meanwhile,treatment with hUC-MSCs iT ameliorated right ventricular hypertrophy(for Fulton’s index,P<0.01),and relieved reduced nephrogenic zone width(P<0.01)and glomerular diameter(P<0.001)in kidneys.Among the beneficial effects,a reduction of BALF protein,MPO,and MDA was observed in hUC-MSCs groups(P<0.01,P<0.001,and P<0.05 respectively).Increased pro-inflammatory cytokines tumor necrosis factor-alpha,interleukin(IL)-1β,and IL-6 expression observed in the hyperoxia group were significantly attenuated by hUC-MSCs administration(P<0.01,P<0.001,and P<0.05 respectively).In addition,we observed an increase in anti-inflammatory cytokine IL-10 expression in rats that received hUC-MSCs iT compared with rats reared in hyperoxia(P<0.05).Transcriptomic analysis showed that the DEGs in lung tissues induced by hyperoxia were enriched in pathways related to inflammatory responses,epithelial cell proliferation,and vasculature development.hUC-MSCs administration blunted these hyperoxia-induced dysregulated genes and resulted in a shift in the gene expression pattern toward the normoxia group.hUC-MSCs increased heme oxygenase-1(HO-1),JAK2,and STAT3 expression,and their phosphorylation in the lung,heart,and kidney(P<0.05).Remarkably,no significant difference was observed between the iT and iP administration.CONCLUSION iT hUC-MSCs administration ameliorates hyperoxia-induced lung,heart,and kidney injuries by activating HO-1 expression and JAK/STAT signaling.The therapeutic benefits of local iT and iP administration are equivalent.