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ObjectiveTo investigate the mechanism of Gegen Qinliantang（GQT） on the intestinal flora of antibiotic-associated diarrhea（AAD） by 16S rRNA sequencing and network pharmacology. MethodSixty SD rats were randomly divided into six groups（n=10）， including blank group， model group， GQT high-， medium- and low-dose groups（10.08， 5.04， 2.52 g·kg-1） as well as Lizhu Changle group（0.15 g·kg-1）， except for the blank group， each group was given clindamycin（250 mg·kg-1） by gavage once a day for 7 consecutive days. After successful modeling， the blank group and the model group were given equal volumes of normal saline by gavage. The other groups were given corresponding doses of drugs by gavage for 14 days. Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP） was used to screen the active components and targets of GQT， GeneCards， Online Mendelian Inheritance in Man（OMIM） database， Pharmacogenetics and Pharmacogenomics Knowledge Base（PharmGKB）， DrugBank and DisGeNET were used to search for AAD disease targets. The drug-disease common targets were obtained by R software. STRING was applied to analyze the target protein-protein interaction， and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway enrichment analysis was performed. Then hematoxylin-eosin（HE） staining was used to observe the pathological changes of the colon， and 16S rRNA sequencing of AAD colon content flora structure further verified the results of network pharmacology. ResultThrough network pharmacology， it was found that 238 active components were screened from GQT and acted on 276 component targets， among which quercetin， puerarin， wogonin and apigenin were the main core components of GQT， 1 097 AAD disease targets and 127 drug-disease intersection targets. The protein-protein interaction network mainly included core targets such as protein kinase B1（Akt1）， interleukin（IL）-6 and IL-1β， which were mainly enriched in the IL-17 signaling pathway. It was verified through animal experiments that compared with the blank group， the colon structure of the model group was seriously abnormal， the intestinal epithelial columnar cells were damaged， the goblet cells were reduced， and a large number of inflammatory cells were infiltrated. Compared with the model group， the colon structure of the GQT high-dose group improved， but there were still abnormalities， the colon structure of GQT medium- and low- dose groups and Lizhu Changle group improved significantly and reached the normal level. GQT could improve the structural diversity of AAD intestinal flora. At the phylum level， the abundance of Firmicutes was increased and the abundance of Bacteroidetes was decreased. At the genus level， the abundance of Lactobacillus was increased， and the abundances of Prevotella and Bacteroides were decreased. Among them， Lactococcus could be used as a biomarker for AAD treatment with GQT， and the prediction of functional metabolism of intestinal flora revealed that GQT could promote acetate and lactate metabolic pathways in the intestine. ConclusionGQT may activate IL-17 signaling pathway by acting on the targets of Akt1 and IL-6 through key components such as quercetin and wogonin， and improve the abundance of Lactococcus in the intestinal tract as well as acetate and lactate metabolic pathways， so as to play a role in repairing the intestinal barrier for the treatment of AAD.
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ObjectiveTo explore the mechanism of Si Junzitang in regulating the expression of NKG2A to affect the anti-colon cancer function of natural killer （NK） cells. MethodNK cells isolated from healthy honors were cultured and used to construct the three incubation models of NK cells， human colon cancer HCT116 cells， and NK cells + HCT116 cells （co-incubation）. real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was conducted to determine the mRNA levels of natural killer group 2 member A （NKG2A） and interleukin （IL）-15 in NK cells， as well as the mRNA level of histocompatibility leucocyte antigen E （HLA-E） in HCT116 cells. The secretion of IL-15 was detected by enzyme-linked immunosorbent assay （ELISA）. Methyl thiazolyl tetrazolium （MTT） assay was employed to determine the applicable concentration of IL-15 and test the effects of Si Junzitang and IL-15 on the activities of NK cells and the HCT116 cells in the co-incubation model. The effects of Si Junzitang and IL-15 on the mRNA levels of NKG2A in NK cells and HLA-E in HCT116 cells were detected by Real-time PCR. Monalizumab （M， anti-NKG2A mab） was used to block the NKG2A-HLA-E pathway in co-incubation model， and then the proliferation of HCT116 cells was detected by MTT assay. ResultThe interaction of NK cells and HCT116 cells up-regulated the mRNA levels of NKG2A in NK cells and HLA-E in HCT116 cells （P<0.05）， as well as the expression level and secretion of IL-15 （P<0.05）. Compared with the blank group， Si Junzitang and Si Junzitang + IL-15 promoted the proliferation and improved the anti-colon cancer function of NK cells （P<0.01）. Furthermore， they down-regulated the mRNA levels of NKG2A in NK cells and HLA-E in the HCT116 cells co-incubated with NK cells （P<0.01）. M and IL-15 + M inhibited the proliferation of HCT116 cells compared with the groups without M （P<0.01）. ConclusionThe interaction of NK cells and HCT116 cells can induce activation of NKG2A-HLA-E pathway to impair NK cell function. Si Junzitang can inhibit the activation of NKG2A-HLA-E pathway to restore the anti-colon cancer function of NK cells.
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ObjectiveTo determine the therapeutic effect of Gegentang granules on a disease-syndrome mouse model combining human coronavirus 229E （hCoV-229E） pneumonia with Hanshi Yidu Xifei syndrome in vivo. MethodMice were randomly divided into normal group， infection group， cold-dampness group， model group， chloroquine phosphate group （0.18 g·kg-1）， interferon-α2b （IFN-α2b） group （1.83×106 U·kg-1）， Gegentang granules high-dose and low-dose groups （6.6， 3.3 g·kg-1） with 10 mice in each group. Cold-dampness environment and hCoV-229E infection were used for modeling， and the general status and lung index of mice in each group were observed. The viral load in lung tissue was detected by real-time fluorescent quantitative polymerase chain reaction （Real-time PCR）， the pathological changes in lung tissue were evaluated by hematoxylin-eosin （HE） staining， the levels of serum gastrointestinal hormones and inflammatory factors in lung tissue were detected by enzyme-linked immunosorbent assay （ELISA）， and the percentage of peripheral blood lymphocytes was detected by flow cytometry. ResultComparing with model group， Gegentang granules could significantly alleviate the physical signs of Hanshi Yidu Xifei syndrome， including listlessness， weakness of limbs， sticky stool， etc. Comparing with model group， Gegentang granules high-dose group significantly reduced lung index， histopathological score of interstitial lung and bronchus， and the level of serum motilin （P<0.05， P<0.01）， two doses of Gegentang granules could significantly increase the level of serum gastrin （P<0.05， P<0.01）， the percentage of CD4+， CD8+ T lymphocytes in peripheral blood （P<0.05， P<0.01）， and the level of tumor necrosis factor-α （TNF-α） in lung tissue was significantly decreased （P<0.01）， and the level of interleukin-6 （IL-6） showed decreasing tendency. ConclusionGegentang granules has therapeutic effect on model mice. It can improve the appearance and behavior characterization， regulate the level of gastrointestinal hormones， decrease lung index and histopathological score， and possibly play an immunomodulatory role by inhibiting the expression of inflammatory cytokines in lung tissue and restoring the percentage of peripheral blood lymphocytes.
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ObjectiveTo determine the therapeutic effect of Gegentang granules on a disease-syndrome mouse model combining human coronavirus 229E （hCoV-229E） pneumonia with Hanshi Yidu Xifei syndrome in vivo. MethodMice were randomly divided into normal group， infection group， cold-dampness group， model group， chloroquine phosphate group （0.18 g·kg-1）， interferon-α2b （IFN-α2b） group （1.83×106 U·kg-1）， Gegentang granules high-dose and low-dose groups （6.6， 3.3 g·kg-1） with 10 mice in each group. Cold-dampness environment and hCoV-229E infection were used for modeling， and the general status and lung index of mice in each group were observed. The viral load in lung tissue was detected by real-time fluorescent quantitative polymerase chain reaction （Real-time PCR）， the pathological changes in lung tissue were evaluated by hematoxylin-eosin （HE） staining， the levels of serum gastrointestinal hormones and inflammatory factors in lung tissue were detected by enzyme-linked immunosorbent assay （ELISA）， and the percentage of peripheral blood lymphocytes was detected by flow cytometry. ResultComparing with model group， Gegentang granules could significantly alleviate the physical signs of Hanshi Yidu Xifei syndrome， including listlessness， weakness of limbs， sticky stool， etc. Comparing with model group， Gegentang granules high-dose group significantly reduced lung index， histopathological score of interstitial lung and bronchus， and the level of serum motilin （P<0.05， P<0.01）， two doses of Gegentang granules could significantly increase the level of serum gastrin （P<0.05， P<0.01）， the percentage of CD4+， CD8+ T lymphocytes in peripheral blood （P<0.05， P<0.01）， and the level of tumor necrosis factor-α （TNF-α） in lung tissue was significantly decreased （P<0.01）， and the level of interleukin-6 （IL-6） showed decreasing tendency. ConclusionGegentang granules has therapeutic effect on model mice. It can improve the appearance and behavior characterization， regulate the level of gastrointestinal hormones， decrease lung index and histopathological score， and possibly play an immunomodulatory role by inhibiting the expression of inflammatory cytokines in lung tissue and restoring the percentage of peripheral blood lymphocytes.
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Objective To evaluate the effect of bone marrow mesenchymal stem cell (BMSC) on the expression of interleukin (IL)-10 and tumor necrosis factor (TNF)-α in mice with ischemia-reperfusion acute kidney injury (IR-AKI). Methods All mice were randomly divided into the sham operation group (control group), ischemia-reperfusion injury group (IRI group) and BMSC treatment group (BMSC group), with 6 mice in each group, respectively. The renal function and pathological changes of mice were detected. The cell apoptosis of renal tissues of mice was determined. The expression levels of serum IL-10 and TNF-α of mice were quantitatively measured. The mouse BMSC was randomly divided into the control and hypoxia-reoxygenation groups (IRI group), and the expression levels of IL-10 and TNF-α in cell supernatant were determined. Results The renal structure of mice was normal in the control group, severe damage was observed in the IRI group, and mild damage occurred in the BMSC group. Compared with the control group, the renal tissue injury scores were significantly higher in the IRI and BMSC groups (both P < 0.05). Compared with the IRI group, the renal tissue injury score was significantly lower in the BMSC group (P < 0.05). Compared with the control group, the levels of serum creatinine (Scr) and blood urea nitrogen (BUN) were remarkably up-regulated in the IRI group, and the level of BUN was significantly up-regulated in the BMSC group (all P < 0.05). Compared with the IRI group, the levels of Scr and BUN were significantly down-regulated in the BMSC group (both P < 0.05). In the IRI group, the quantity of apoptotic cells in the renal tissues was considerably higher than those in the BMSC and control groups, and the quantity of apoptotic cells in the BMSC group was significantly higher than that in the control group (all P < 0.05). Compared with the control group, the levels of serum IL-10 and TNF-α were significantly up-regulated in the IRI group, whereas the level of serum TNF-α was significantly down-regulated and the level of serum IL-10 was significantly up-regulated in the BMSC group (all P < 0.05). Compared with the IRI group, the levels of serum IL-10 and TNF-α were significantly down-regulated in the BMSC group (both P < 0.05). The levels of IL-10 and TNF-α in the cell supernatant did not significantly differ between the IRI and control groups (P=0.080、0.627). Conclusions BMSC infusion may reduce the incidence of renal IRI and inflammation, probably via the mechanism of down-regulating TNF-α expression rather than up-regulating IL-10 expression.
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Objective To investigate the effect and its molecular mechanism of phosphoglycerate mutase 5 (PGAM5) mediated pyroptosis on liver ischemia-reperfusion injury (IRI). Methods C57 mouse models of liver IRI were established and randomly divided into the 6 h reperfusion (6 h group) and 12 h reperfusion (12 h group), and sham operation group (sham group) was established too, 10 rats in each group. The effect of IRI on the parameters in the liver tissues and serum samples was evaluated. The expression levels of PGAM5 and cysteinyl aspartate specific proteinase (Caspase)-1 in the liver tissues during IRI were quantitatively detected. The IRI models of liver cells were established (IRI group). The IRI models of liver cells were established after pretreatment with Caspase-1 inhibitor Z-YVAD-FMK (inhibitor group). The untreated AML12 cells were allocated into the control group. The effect of inhibiting Caspase-1 activity on pyroptosis was analyzed. AML12 cells were transfected with PGAM5 small interfering ribonucleic acid (siRNA) (siRNA group) and siRNA-negative control (siRNA-NC) (siRNA-NC group) by liposome 3000, and then IRI models of liver cells were established. The untreated AML12 cells were assigned into the control group. The effect of PGAM5 mediated pyroptosis on IRI of liver cells was assessed. Results In the 6 h and 12 h groups, partial liver cell edema, hepatic sinusoid narrowing, central vein congestion and occasional spot necrosis were observed in the mouse liver tissues, and these changes in the 12 h group were more aggravated than those in the 6 h group. The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the 6 h and 12 h groups were higher than those in the sham group, and the values in the 12 h group were higher than those in the 6 h group. The levels of tumor necrosis factor (TNF)-α and interleukin (IL)-1β were increased in the 6 h and 12 h groups, and the values in the 12 h group were lower than those in the 6 h group. The relative expression levels of IL-1β messenger ribonucleic acid (mRNA) in the mouse liver tissues in the 6 h and 12 h groups were up-regulated, and the value in the 12 h group was lower than that in the 6 h group. The cell apoptosis rates in the liver tissues were significantly increased in the 6 h and 12 h groups, and the value in the 12 h group was remarkably lower than that in the 6 h group (P < 0.01-0.05). Compared with the sham group, the relative expression levels of PGAM5 mRNA and protein in the mouse liver tissues in the 6 h and 12 h groups were significantly up-regulated, and the values in the 12 h group were significantly higher than those in the 6 h group (P < 0.01-0.05). The protein expression levels of PGAM5 and Caspase-1 in the liver tissues were up-regulated in the 6 h and 12 h groups. Compared with the control group, the relative expression levels of NOD-like receptor protein 3 (NLRP3), cleaved Caspase-1 and Gasdermin D (GSDMD) proteins were up-regulated and the fluorescence intensity of GSDMD was increased in the IRI group. Compared with the IRI group, the relative expression levels of NLRP3, cleaved Caspase-1 and GSDMD proteins were significantly down-regulated and the fluorescence intensity of GSDMD was considerably decreased in the inhibitor group (P < 0.01-0.05). Compared with the control group, the cell survival rate was significantly decreased, and the relative expression levels of PGAM5, NLRP3, cleaved Caspase-1 and GSDMD proteins were significantly up-regulated in the siRNA-NC group (P < 0.01-0.05). Compared with the siRNA-NC group, the cell survival rate was remarkably increased, whereas the relative expression levels of PGAM5, NLRP3, cleaved Caspase-1 and GSDMD proteins were significantly down-regulated in the siRNA group (P < 0.01-0.05). Conclusions PGAM5 may aggravate the liver IRI in mouse models probably by mediating pyroptosis via PGAM5/Caspase-1/GSDMD signaling pathway and aggravating liver cell injury.
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With the improvement of people's living standard and the change of dietary structure, the prevalence of gout has increased gradually with the increased intake of protein, sugar and fat. There has been a positive correlation between gout and age, and the age of onset decreased gradually. The inflammation induced by sodium urate crystal is the pathological basis of gout, which activates innate immunity, releases many kinds of inflammatory mediators, such as interleukin(IL)-1β, IL-6 and tumor necrosis factor(TNF)-α, and then causes inflammatory cascade reaction and acute attacks, such as joint redness, swelling and heat pain. There is a spontaneous remission mechanism in gout. For one thing, macrophages reduce the stimulation of monosodium urate(MSU) through phagocytosis of MSU crystals as foreign bodies, for another, differentiated and mature macrophages secrete anti-inflammatory factor transforming growth factor(TGF)-β1, inhibit the expression of inflammatory factors and promote spontaneous relief of acute gout attack. In addition to the activation mechanism of intracellular signaling molecules associated with inflammatory response, the inflammatory mechanism of gout also involves complement activation, cell activation and other pathways. The complications caused by gout, such as cardiovascular system damage and joint destruction, are seriously harmful to human health. At present, western drugs, such as allopurinol and febuxostat, exert an effect in inhibiting xanthine oxidase. Benzimarone has effect in reducing renal absorption of uric acid and promoting uric acid excretion by inhibiting uric acid transporter 1(URAT1) and glucose transporter 9(GLUT9). Even Lesinurad and other medicines in current studies are based on the inhibition of uric acid re-absorption, but with adverse reactions that limit the clinical application. The treatment of gout with traditional Chinese medicine(TCM) has multi-target characteristics, with advantages in reducing uric acid, resisting inflammation and improving joint function and a high safety. It has been gradually popularized and applied in clinical treatment of gout. Therefore, it is a promising research direction to treat gout with TCM and western medicine based on the pathomechanism of gout.
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Objective:To investigate the relationship between NOD-like receptor pyrin domain containing 3(NLRP3)/cysteine aspartate-specific protease(Caspase)-1 signaling pathway and esophageal inflammation by observing the effect of Xuanfu Daizhe Tang on the composition of inflammatory body and the expression of relevant inflammatory factors in rats with reflux esophagitis (RE), so as to explain the mechanism of Xuanfu Daizhe Tang in treating RE. Method:Sixty healthy male Wistar rats were randomly divided into four groups:the normal control group, the model control group, the Xuanfu Daizhe Tang group (9.89 g·kg-1) and the positive control group (omeprazole enteric-coated tablets+mosapride, 2.58 mg·kg-1), with 15 rats in each group. Except for the blank control group, the remaining rats were operated by " 4.2 mm pyloric clip+2/3 gastric fundus ligation" to establish models. Since the 8th day after the operation, the rats were given corresponding drugs twice a day for 14 days. The arterial blood and esophageal tissues were taken out at the 15th day after the intervention. The pathological morphology of esophagus was observed by naked eyes and under light microscopy. The secretion of cytokines Caspase-1 and interleukin(IL)-1β in serum was detected by enzyme linked immunosorbent assay(ELISA). The expressions of NLRP3, Caspase-1 and IL-1β in esophagus were detected by Western blot. Result:Compared with the normal group, the injury of esophageal mucosa in the model group was the most serious. Compared with the normal group, the levels of Caspase-1 and IL-1β in serum and the expression of NLRP3 protein in esophageal tissue of the model group were significantly increased (PPβ in serum of rats, and down-regulate the expressions of NLRP3, Caspase-1 and IL-1β protein in esophageal tissue (P0.05, PConclusion:Xuanfu Daizhe Tang can regulate the expressions of NLRP3 and Caspase-1, and reduce the content of IL-1β, suggesting that it may antagonize esophageal inflammatory response, reduce esophageal inflammatory injury and treat RE by inhibiting the activation of NLRP3/Caspase-1 signaling pathway.
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Objective@#To determine the changes in peripheral plasma concentrations of interleukin-10 (IL-10), interleukin -12 (IL-12) and interfoeron-γ(IFN-γ) in the patients with chronic hepatitis B virus (HBV) infection and their correlations with HBV infection stage or HBV DNA load of HBV carriers.@*Methods@#Data of 135 patients with chronic HBV infection from March 2016 to March 2017 were collected, the patients included 32 chronic HBV carriers, 61 with chronic hepatitis and 42 with cirrhosis. Forty healthy subjects served as controls. The concentrations of IL-10, IL-12 and IFN-γ were determined using enzyme-linked immunosorbent assay (ELISA). Correlation analysis was performed using the Pearson correlation test, which was performed to analyze the correlation between IL-10, IL-12, IFN-γ and HBV infection stage, HBV DNA load of HBV carriers.@*Results@#Compared with those in healthy controls, plasma IL-10 and IL-12 levels in patients with chronic hepatitis and cirrhosis increased significantly (F=22.06, 15.67, P=0.013, 0.021), plasma IL-10 and IL-12 levels in cirrhosis cases were higher than those in chronic hepatitis (all P<0.001), plasma IL-10 and IL-12 levels in chronic hepatitis were higher than those in chronic HBV carriers (all P<0.001). Plasma IFN-γ level in chronic HBV carriers, chronic hepatitis and cirrhosis were significantly higher than those in healthy controls (F=18.36, P=0.017). There were statistically significant differences in IFN-γ levels among the three groups in the chronic HBV carriers, chronic hepatitis and cirrhosis. IL-10, IL-12 and IFN-γ levels of the low, medium and high HBV DNA load groups were statistically significant (all P<0.05). There was no correlation between IL-10 and HBV DNA. IFN-γ, IL-12 and HBV DNA load were negatively correlated. There was no correlation between IL-10 and IFN-γ (r=0.103, P>0.05), IL-12 and IFN-γ were significantly positively correlated (r=0.687, P<0.05).@*Conclusions@#IL-10, IL-12 and IFN-γ may play an important role in the chronic HBV infection.
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The effects of diets containing different concentration of Saccharina japonica algae (0%, 5%, 10%, 15%, 20% and 25%) on growth and Interleukin (IL)-10 gene expression of juvenile sea cucumber Apostichopus japonicus were studied. At first, 08 weeks feeding trail was conducted to evaluate the growth performance of sea cucumbers fed with one of the six experimental diets. Result showed that sea cucumbers fed 15% Saccharina japonica algae diet had higher specific growth rate (SGR) and food conversion efficiency (FCE) than the other experimental diets (P<0.05). Secondly, Interleukin (IL)-10 gene expression was determined where mice splenocytes were treated with different experimental diets fed sea cucumber extracts for two hours. The highest Interleukin (IL)-10 gene expressions was found in 15% Saccharina japonica algae diets fed sea cucumbers extract compared to other diets except 10% Saccharina japonica algae diet. Results of this experiment suggest that 15% Saccharina japonica containing diet perform better growth and could elevate IL-10 gene expression. This information might be useful in the further development of more appropriate diets for the culture of sea cucumbers.
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PURPOSE: Podocytes are important architectures that maintain the crucial roles of glomerular filtration barrier functions. Despite this structural importance, however, the mechanisms of the changes in podocytes that can be an important pathogenesis of minimal change nephrotic syndrome (MCNS) are not clear yet. The aim of this study was to investigate whether apoptosis is induced by interleukin (IL)-13 in cultured human podocytes. METHODS: Human podocytes were treated with different IL-13 doses and apoptotic cells were analyzed using terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL assay) and fluorescence-activated cell sorting (FACS). RESULTS: The IL-13 increased the number of TUNEL-positive cells in a dose-dependent manner at 6 and 18 hours (P<0.05 and P<0.05, respectively). The apoptosis rate was appeared to be increased slightly in the IL-13-stimulated podocytes (8.63%, 13.02%, and 14.46%; 3, 10 and 30 ng/mL, respectively) than in the control cells (7.66%) at 12 hours by FACS assay. CONCLUSION: Our study revealed that IL-13 expression may increase podocyte apoptosis. Blocking the IL-13 signal pathway can potentially play an important role in regulating the apoptosis of podocytes.
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Humans , Apoptosis , DNA Nucleotidylexotransferase , Flow Cytometry , Glomerular Filtration Barrier , Interleukin-13 , Interleukins , Nephrosis, Lipoid , Podocytes , Signal TransductionABSTRACT
Objective To investigate the effect of human umbilical cord mesenchymal stem cells (HUC-MSCs) on CD4+T cells in liver after hepatic ischemia-reperfusion injury (HIRI) in mice. Methods Two hundred and twenty-five mice were randomly divided into sham group, control group and MSC group, with 75 mice in each group. HIRI model mice were used in MSC group and control group. HUC-MSCs were injected in MSC group through inferior vena cava. Normal saline was injected in control group through inferior vena cava. Only laparotomy and abdominal closure were performed in sham group without blood vessel clipping. At 6, 12 and 24 h after operation, 15 mice of each group were randomly selected to sample eyeball blood and liver tissues, and the 30 mice left in each group were used to extract intrahepatic mononuclear cells. The number of intrahepatic mononuclear cells, percentage, number and positive rate of CD4+T cells in the mice of various groups at different time points were compared. The content of interleukin (IL)-17 in serum and liver tissue as well as expression levels of costimulatory molecules B7-1 and B7-2 messenger RNA (mRNA) in liver tissues of the mice at different time points were compared. Results At 12 and 24 h after operation, the number of intrahepatic mononuclear cells of control group was significantly higher than that of sham group, while the number of intrahepatic mononuclear cells of MSC group was significantly lower than that of control group (P<0.01-0.05). At 6, 12 and 24 h after operation, the percentage, number and positive rate of CD4+T cells of control group were significantly higher than those of sham group (all P<0.01), while the percentage of CD4+T cells of MSC group was significantly lower than that of control group (P<0.01-0.05). At 12 and 24 h after operation, the number and positive rate of CD4+T cells of MSC group were significantly lower than those of control group (P<0.01-0.05). At 6, 12 and 24 h after operation, the IL-17 contents in serum and liver tissues of control group were higher than those of sham group (all P<0.01), while the IL-17 contents in serum and liver tissues of MSC group were lower than those of control group (all P<0.01). At 6 h after operation, the mRNA expression level of B7-2 of control group was higher than that of sham group (P<0.05). At 12 and 24 h after operation, the mRNA expression levels of B7-1 and B7-2 of control group were higher than those of sham group (all P<0.01), while the mRNA expression levels of B7-1 and B7-2 of MSC group were lower than those of control group (all P<0.01). Conclusions HUC-MSCs inhibits the number of CD4+T cells and the secretion of IL-17 in liver after HIRI, as well as decreases the number of intrahepatic mononuclear cells and the mRNA expression of B7-1 and B7-2, thereby alleviating HIRI.
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Objective To study the biological behaviors and effects of immunoglobulin-like transcript-4 (ILT4) expression in mononuclear cells on the prognosis of sepsis.Methods ILT4 +/+ (WT) and ILT4-knockout mice (ILT4-/-) male BALB/c mice were used for sepsis modeling using cecal ligation puncture (CLP).Flow cytometry was used to measure the levels of expression of ILT4 and major histocompatihility complex class Ⅱ molecules (MHC-Ⅱ) in mononuclear cells of peripheral blood 24 h after CLP.ELISA was used to measure the concentrations of interleukin-6 (IL-6) and serum tumor necrosis factor-alpha (TNF-α) in different groups of mice at 0 h,6 h,12 h,and 24 h after CLP to monitor the survival and prognosis over the course of 168 h.Results ILT4 was highly expressed in mononuclear cells of the peripheral blood of septic mice 24 h after CLP in comparison with that before CLP (1292.00 ± 143.70) vs.(193.50 ± 52.54),P ＜ 0.05.MHC-Ⅱ expression in mononuclear cells of the peripheral blood in ILT4-/-mice was significantly higher than that in WT mice (49.38 ± 5.66)％ vs.(24.25 ± 6.76) ％,P ＜ 0.05).Serum IL-6 was significantly elevated 24 h after CLP compared with that before CLP (470.75 ± 88.03) vs.(54.25 ± 20.04),P ＜ 0.05.The serum IL-6 concentration was much lower in ILT4-/-mice thanthatin MT mice (241.25 ± 45.10)vs.(470.75 ± 88.03),P ＜ 0.05;whereas,there was no significant difference in TNF-α expression between two groups of mice (50.88 ± 6.38) vs.(53.13 ± 5.49),P ＞ 0.05.The survival rate of ILT4-/-mice was significantly higher after CLP compared with WT mice (P ＜ 0.05).Conclusion The high level of ILT4 expression in mononuclear cells were observed in peripheral blood during sepsis and it was found to be associated with high serum IL-6 levels and low MHC-Ⅱ expression in mononuclear cells,leading to increased mortality.
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Objective To detect the levels of helper T cells (Th) 17 and Th9 cells in the peripheral blood of patients with pedi-atric bronchial asthma and the expression of interleukin 17 (IL-17) ,IL-9 and total IgE in serum ,and to discuss their effect and clini-cal significance in pathogenesis of pediatric asthma .Methods A total of 46 children with bronchial asthma ,including asthma attack group (n=26) and asthma remission group (n=20) were selected .Healthy children were selected as healthy control group (n=20) .Flow cytometry was used to detect the percentages of Th17 and Th9 cells in the peripheral blood .The levels of IL-17 and IL-9 in serum were measured by enzyme-linked immunosorbent assay (ELISA) .Serum total IgE levels were measured by the specific protein analyzer .Results The percentages of Th17 and Th9 cells in the peripheral blood were significantly higher in asthmatic group compared with remission group and healthy control group (P<0 .05) .The percentages of Th17 and Th9 cells in the periph-eral blood was significantly higher in remission group compared with healthy control group (P<0 .05) .The levels of IL-17 ,IL-9 and total IgE in serum were significantly higher in asthmatic group compared with remission group and healthy control group (P<0 .05) .The levels of IL-17 ,IL-9 and total IgE in serum were significantly higher in remission group compared with healthy control group (P<0 .05) .The levels of IL-17 and IL-9 in serum of asthmatic children were positively correlated with total IgE levels (P<0 .05) .The correlation analysis results showed that the levels of IL-17 ,IL-9 and total IgE in serum of patients with pediatric bron-chial asthma have a positive correlation .(r=0 .717 ,0 .491 ,0 .786 ,P<0 .05) .Conclusion Th17 ,Th9 cells and their cytokines in pe-ripheral blood of patients with pediatric bronchial asthma play important roles in the pathogenesis of asthma and are positively cor-related with the severity of asthma .
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Objective To explore the clinical value of serum interleukin(IL)-33 and its soluble receptor ST2(sST2) level in patients with chronic hepatitis B.Methods A total of 65 cases with chronic hepatitis B were recruited into study group,meanwhile 60 healthy persons were enrolled in the control group from January 2014 to October 2016 in our hospital.IL-33,sST2 and alanine aminotransferase(ALT) were detected and compared in the two groups.Results The level of IL-33,sST2 and ALT were significant higher than those of the control group(t=6.542,7.218,6.324;P<0.05).IL-33 and sST2 levels in chronic hepatitis B patients with abnormal ALT level were significant higher than those with normal ALT(t=16.328,9.874,P<0.05).Conclusion The detection of IL-33 and sST2 in patients with chronic hepatitis B could help to understand the immune status of patients,and provide a theoretical basis for the treatment of chronic hepatitis B.
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CONTEXT: Trichosanthin produced in the root tube of Trichosanthes kirilowii shows anti-tumor activity on a series of cancer cells including Hela, MCF-7, HL-60. But there is little information about its effect on the carcinogenesis of prostate cancer. OBJECTIVE: This work was designed to study the role of trichosanthin on prostate cancer cells PC3. MATERIALS AND METHODS: Trichosanthin was expressed in BL21 strain and purified by affinity chromatography. MTT assay was designed to determine the effect of trichosanthin on growth of PC3 cells at doses of 10, 20, 40, 60, 80, and 120 µg/ml.Then the effect of 50 µg/ml rTCS alone or combined with 2 µM IL-2 on PC3 cell proliferation was analyzed. And the mechanism of rTCS was studied by western blot. After that the in vivo effect of rTCS combined with IL-2 was explored in mice bearing PC3 xenograft tumor. RESULTS: Trichosanthin was successfully expressed in BL21 and purified by 100 mM imidazole. It was shown to inhibit proliferation of PC3 cells in a dose-dependent manner with IC50 50.6 µg/ml. When combined with cytokine IL-2, a significant synergic effect was obtained. The inhibition rate on PC3 was around 50 % in combination group while only 35.5 % in single rTCS group at 50 µg/ml. Further, the expression of full length caspase-8 and Bcl-2 decreased significantly while cleaved caspase-8 and Bax were up-regulated, which suggest that caspase-8-mediated apoptosis pathway may be activated by rTCS in PC3 cells. Moreover, our data demonstrated that tumor volume and tumor weight were significantly reduced in rTCS-treated or rTCS/IL-2-treated nude mice bearing PC3 xenograft tumor compared with control. And significant difference was also found between rTCS and rTCS/IL-2 group. CONCLUSIONS: This study demonstrates that rTCS is a potential agent with high in vitro and in vivo anti-tumor activity on PC3 cells. And rTCS combined with IL-2 is a promising strategy in treating patients with prostate cancer in future.
Subject(s)
Animals , Male , Female , Mice , Prostatic Neoplasms/drug therapy , Trichosanthin/pharmacology , Antineoplastic Agents, Phytogenic/pharmacology , Prostatic Neoplasms/pathology , Tetrazolium Salts , Time Factors , Recombinant Proteins/pharmacology , Blotting, Western , Reproducibility of Results , Apoptosis/drug effects , Cell Line, Tumor , Tumor Burden , Cell Proliferation/drug effects , FormazansABSTRACT
Objective:To explore the association between interleukin(IL)-28B single nucleotide polymorphisms and natural outcome of hepatitis C virus.Methods:The IL-28B rs12979860 locus was genotyped in 266 HCV infected volunteer blood donors(107 spontaneous cleared and 159 chronic infection) and 97 healthy controls using Sanger sequencing assay.The difference in rs12979860 genotypes and allele frequencies between the six groups(107 spontaneous cleared and 159 chronic infection,266 HCV infection and 97 healthy controls,159 chronic infection and 97 healthy controls) were analyzed by statistics.Results:159 HCV chronic infection,107 spontaneous cleared and 97 healthy controls,were shown more CC genotype,accounting for 83.6%,95.3%and 86.6%,respectively, while the CT genotype accounted for 16.4%,4.7%and 13.4%respectively.No TT genotype was found.The CC/CT genotype was not significant difference between HCV infection and healthy controls,chronic infection and healthy controls(χ2=0.204,P=0.652;χ2=0.406,P=0.524),but between chronic infections and spontaneous clearance had statistically significant(χ2=8.474,P=0.004),the frequence of C allele in spontaneous cleared was higher than HCV chronic infection(χ2=7.949,P=0.005).Conclusion: The gene polymorphism of IL-28B rs12979860 is not related to HCV susceptibility,but there are differences in chronic infection and spontaneous cleared,showing the C allelic in favor of HCV spontaneous cleaed.
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Accumulating evidences indicate that some diseases are triggered by abnormalities of the gut microbiota. Among these, immune-related diseases can be the promising targets for probiotcs. Several studies have proved the efficacy of probiotics for preventing such diseases including cancers, infections, allergies, inflammatory bowel diseases and autoimmune diseases. Lactobacillus casei strain Shirota (LcS) is one of the most popular probiotics, benefits of which in health maintenance and disease control have been supported by several science-based evidences. This review summarizes human clinical trials with this probiotic against cancer development and also discusses the possible immunomodulatory mechanisms by which LcS exerts anti-cancer activity.
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Entamoeba histolytica produces Monocyte Locomotion Inhibitory Factor (MLIF), which may contribute to the delayed inflammation observed in amoebic hepatic abscesses. Leukocytes are affected through the modulation of cytokine expression and/or production. We evaluated the effects of MLIF on the activation and production of intracellular cytokines in human CD4+ T lymphocytes by flow cytometry. Cells were stimulated for 24 h with PMA, MLIF, or PMA+MLIF. Cellular activation was measured using anti-CD69. Th1/Th2 production was studied by the expression of intracellular cytokines and cytokine/chemokine receptors. MLIF increased CD69 and induced the over-expression of the IL-l©¬, IFN-¥ã, IL-2, IL-4, and IL-10 intracellular cytokines; PMA+MLIF inhibited Th1 cytokine (IFN-¥ã) and increased Th2 cytokines (IL-4 and IL-10). The co-expression of the cytokine and chemokine receptors IFN-¥ã/CCR5 and IL-1©¬/CCR5 was inhibited by PMA+MLIF and Th2 co-expression was increased. MLIF effects varied depending on the conditions. MLIF alone activated the Th1 and Th2 cytokines and cytokine/receptor expression; however, PMA+MLIF increased the expression of Th2 but inhibited it in Th1.
Subject(s)
Female , Humans , Male , Cytokines/biosynthesis , Oligopeptides/pharmacology , /drug effects , /drug effects , Th1 Cells/drug effects , /drug effects , Cells, Cultured , Entamoeba histolytica/immunology , Flow Cytometry , Oligopeptides/biosynthesis , /immunology , /immunology , Tetradecanoylphorbol Acetate/pharmacology , Th1 Cells/immunology , /immunologyABSTRACT
@#Objective To investigate the effect of erythropoietin(EPO)as a brain-protective factor and inflammatory cytokines in the pathogenesis of cerebral palsy(CP).Methods Serum samples from 31 CP patients,37 neonates with CP risk factors such as hypoxic-ischemic injury and/or perinatal infection,and 20 controls of neonates or children were obtained respectively.EPO,tumor necrosis factor(TNF)-α and interleukin(IL)-6 levels were measured with the enzyme-linked immunosorbent assay double sandwich method(ABC-ELISA).Results The serum EPO level of neonatal patients was higher than that of controls or CP group(P<0.01),but there was no significant difference between CP group and controls.The serum TNF-α and IL-6 levels of CP and neonatal patients were higher than that in controls(P<0.01).The serum TNF-α level of CP group was higher than that of neonatal patients(P<0.05).There was no significant difference between CP group and neonatal patients in serum IL-6 level.Conclusion The inflammatory responses mediated by proinflammatory cytokines may play a role in the pathogenesis of cerebral palsy.