IL-6 Content Research Articles

Стан локального цервікального імунітету в жінок із непліддям

Purpose - to study certain indicators of local immunity in women with infertility of various origins and early cervical lesions Materials and methods. The state of local cervical immunity was examined in 496 women, who were divided into groups depending on the cause of infertility, and each group was divided into subgroups A and B depending on the presence of cervical epithelial lesions. 128 women in the Group I had an endocrine cause, 122 patients in the Group II had a tubal-peritoneal factor of infertility, and 121 patients in the Group III had a male factor of fertility disorder. 125 healthy women formed the control group (CG). Subgroup A included women with NILM liquid cytology results: IA - 100 women, IIA - 70, IIIA - 68, GCA - 88. Subgroups B included women with ASC-US (atypical squamous cells of undetermined significance), LSIL (low-grade squamous intraepithelial lesion) and HSIL (high-grade squamous intraepithelial lesion): IB - 28 patients, IIB - 52, IIIB - 53, CGB - 37. In all patients, the content of interleukin-8 and -6 (IL-8, IL-6), lactoferrin and secretory leukocyte proteinase inhibitor in the material obtained for liquid cytology was determined by polymerase chain reaction. Results. In all patients with infertility, an increased concentration of secretory leukocyte proteinase inhibitor (from 314.1 to 398.7 pg/ml) and a decreased lactoferrin content (from 13.6 to 28.6 pg/ml) were found compared to the group of healthy women (114.8 and 64.8 pg/ml, respectively). In women with tubal and peritoneal infertility and male infertility, higher concentrations of IL-8 were found (659.9 to 878.8 pg/ml) than in the group of endocrine infertility (302.9 pg/ml) and in the group of healthy women (282.4 pg/ml). Conclusions. IL-8 content in cervical mucus is probably higher in women with tubal-peritoneal infertility (838 pg/ml) and male infertility (659.9 pg/ml) than in healthy women (289.4 pg/ml) and in women with endocrine infertility (302.3 pg/ml). The content of secretory inhibitor of leukocyte proteinase in cervical mucus was increased in all patients with infertility, regardless of its cause (from 314.1 pg/ml to 398.9 pg/ml), in the group of healthy women - 144.8 pg/ml. 3. The content of lactoferrin in the mucus of the cervical canal was reduced in all patients with impaired fertility (from 13.6 pg/ml to 28.6 pg/ml), in the group of healthy women - 64.8 pg/ml. The research was carried out in accordance with the principles of the Helsinki Declaration. The study protocol was approved by the Local Ethics Committee of the participating institution. The informed consent of the patient was obtained for conducting the studies. No conflict of interests was declared by the author.

Inhalation of Citrus Reticulata essential oil alleviates airway inflammation and emphysema in COPD rats through regulation of macrophages

Ethnopharmacological relevanceChronic obstructive pulmonary disease (COPD) is a chronic inflammatory respiratory disease. Citrus Reticulata peel, the dried ripe peel of Citrus Reticulata species, has been found to have anti-inflammatory and cough attenuation effects. However, the therapeutic effects and its precise underlying mechanisms of atomizing inhalation using Citrus Reticulata essential oil (CREO) have not yet been fully elucidated. Aim of the studyThe aim of this study was to assess the therapeutic effects of Citrus Reticulata essential oil and its associated anti-inflammatory mechanisms in COPD rat model. MethodsA total of 80 SD rats were randomized into four groups: control group (Con), COPD model group (COPD), COPD + ipratropium bromide (IB), and COPD + citrus reticulata essential oil (CREO). To induce COPD in rats, cigarette smoke (CS) exposure was used, while CREO and IB groups were administered through atomizing inhalation. The clinical signs, pathological lesions of the lung, percentages of antigen-presenting lung macrophages (CD11b/c+/CD86+ cells) and CD8+ T cells, and the content and mRNA expression of cytokines of the lung were analyzed. ResultsThe findings revealed that atomizing inhalation of Citrus reticulata essential oil had therapeutic effects on COPD rats. The treatment resulted in improvement in the body weight and mental status of COPD rats, reduced pathological injury of the lung, and increased proportion of CD11b/c+/CD86+ cells in lung macrophages, while also decreasing the number of CD8+ T cells. In addition, the Citrus Reticulata essential oil reduced the contents of IL-18, IL-17A, IL-12p70, and GM-CSF, downregulated the relative mRNA expression of IFN-γ, IL-4, and MMP-12, and upregulated the mRNA expression of IL-10. ConclusionsCitrus reticulata essential oil can alleviate histological injury of the lung and regulate macrophages and CD8+ T cells in COPD rats. The study suggests that citrus reticulata essential oil could be a potential therapeutic agent for COPD.

Features of innate and adaptive immunity in patients with moderate course of COVID-19 with various degrees of lung tissue damage

Objective: to study changes in innate and adaptive immunity in patients with a moderate course of COVID-19 with varying degrees of lung tissue damage. 40 patients with a moderate form of COVID-19 and varying degrees of damage to the lung tissue (up to 25% and up to 50%) and 30 patients from the control group (healthy donors) were examined using methods: computed tomography of the lungs, evaluation of a complete blood count, biochemical parameters, immunological studies, statistical. Differences in all patients with the control group consisted in a decrease in the absolute content of lymphocytes, in a significant increase in the relative content of natural killers and cytokine imbalance. The most significant differences were registered in the content of IL-6, depending on the degree of damage to the lung tissue. A smaller area of lung damage reflects a milder course of the disease, which does not exclude the need for dispensary observation and rehabilitation after discharge.

Florfenicol can inhibit chick growth and lead to immunosuppression1

Florfenicol (FLO) is a chemically synthesized broad-spectrum antimicrobial agent of amide alcohols for animals, which is one of the most widely used antimicrobials in livestock, poultry, and aquaculture. With the use of FLO, more and more attention has been paid to its hematopoietic toxicity, immunotoxicity, genotoxicity, and embryotoxicity. In this study, SPF chicks at the age of 3 d began to drink water with the FLO at a dose of 100 mg L-1 for six consecutive days, and the growth performance of chicks was monitored, the effect of FLO on immune organs was detected by pathological examination and TUNEL apoptosis staining. In order to evaluate the level of organism immunity, the level of Newcastle disease virus antibody in serum was detected by hemagglutination inhibition test, the content of cytokines (IL-1, IL-2, IL-6, TNF-α, IFN-γ) in serum was detected by ELISA, and the transcription of interferon-related genes (IRF-7, 2’-5’OAS, Mx1) and cytokine genes (IL-6, TNF-α, IFN-γ) in immune organs were detected by real time fluorescence quantitative PCR. The results showed that the early application of FLO could inhibit the growth and development of chicks, and the body weight and immune organ index of the treatment group were lower than those of the control group. Histopathological examination showed that there was a decrease in the number of lymphocytes in the bursa of Fabricius in the treatment group in the early stage of drug withdrawal, and the results of TUNEL apoptosis staining in the bursa of Fabricius showed that obvious lymphocyte apoptosis occurred in the FLO treatment group. Compared with the control group, the transcription levels of interferon-related genes IRF-7, 2’-5’OAS, and cytokine genes IL-6, TNF-α and IFN-γ in FLO treatment group decreased to a certain extent, while the transcription level of Mx1 gene had no significant difference at all time points. The level of serum NDV antibody and the contents of cytokines IL-1, IL-2 and IFN-γ in the FLO treatment group were significantly lower than those in the control group in the early stage of drug withdrawal, but recovered gradually in the later stage. This study showed that FLO has a certain degree of effect on the immune function of chicks, and the results of the study laid the foundation for further research on the mechanism of FLO-induced immunotoxicity.

Rhodotorula mucilaginosa ZTHY2 Attenuates Cyclophosphamide-Induced Immunosuppression in Mice.

Rhodotorula mucilaginosa (R. mucilaginosa) can enhance the immune and antioxidant function of the body. However, whether R. mucilaginosa has an immunoregulatory effect on cyclophosphamide (CTX)-induced immunosuppressed animals remains to be clarified. In this study, the R. mucilaginosa ZTHY2 that we isolated from the coastal waters of the South China Sea previously was prepared in order to investigate its immunoprotective effect on CTX-induced immunosuppression in mice, and the effects were compared to those of Lactobacillus acidophilus (LA) (a well-known probiotic). Seventy-two male SPF mice were divided into six groups: The C group (control); IM group (immunosuppressive model group) (+CTX); Rl, Rm, and Rh groups (+CTX+low, medium, and high concentration of R. mucilaginosa, respectively); and PC (positive control) group (+CTX+LA). After a 28-day feeding trial, blood samples were taken for biochemical and serum immunological analysis, and the thymus and spleen were collected to analyze the organ index, lymphocyte proliferation and differentiation, and antioxidant capacity. The findings showed that R. mucilaginosa ZTHY2 improved the spleen and thymus indices, effectively attenuated immune organ atrophy caused by CTX, and enhanced the proliferation of T and B lymphocytes induced by ConA and LPS. R. mucilaginosa ZTHY2 promoted the secretion of cytokines and immunoglobulins and significantly increased the contents of IL-2, IL-4, IL-6, TNF-α, IFN-γ, IgA, IgG, IgM, CD4, CD8, CD19, and CD20 in serum. The proportion of CD4+, CD8+, CD19+, and CD20+ lymphocytes in spleen, thymus, and mesenteric lymph nodes were increased. In addition, R. mucilaginosa ZTHY2 reduced the reactive oxygen species (ROS) and malondialdehyde (MDA) levels and increased glutathione (GSH), total superoxide dismutase (SOD), and catalase (CAT) levels. Our results indicated that R. mucilaginosa ZTHY2 can significantly enhance the immune function of immunosuppressed mice, and improving antioxidant capacity thus attenuates CTX-induced immunosuppression and immune organ atrophy.

Antitumor effect of exopolysaccharide from Lactiplantibacillus plantarum WLPL09 on melanoma mice via regulating immunity and gut microbiota

Exopolysaccharide (EPS-09) from L. plantarum WLPL09 was systemically investigated for the antitumor effect in B16F10 melanoma bearing mice model. The results showed that administraion of EPS-09 (200 mg/kg) could sigificantly inhibit the tumor growth of melanoma bearing mice, with a inhibition rate of 42.53 %. Meanwhile, compared to the Model group, high dose of EPS-09 (200 mg/kg) administraion could increase the spleen index (P = 0.10), promote the splenic lymphocytes proliferation under the stimulation of ConA and LPS with a proliferation rate of 120.58 % and 169.88 %, respectively, enhance the amount of CD4+ and CD8+ T cells (P < 0.0001, P = 0.0149) in tumor tissue, as well as the serum content of cytokines, i.e., TNF-α, IFN-γ, IL-2 (P < 0.05) and IL-6 (P = 0.039) of B16F10 melanoma bearing mice. The transcriptional level analysis revealed that EPS-09 (200 mg/kg) administraion could sigificantly (P < 0.05) upregulate the transcription of apoptosis raleted genes, i.e., P53, Caspase-3 and Caspase-9, and the ratio of Bax/Bcl-2, downregulate the transcription of angiogenesis markers, i.e., Vegf and Fgf2 compared with Model group. Furthermore, administration of EPS-09 could increase the abundance of phylum Firmicutes, family Ruminococcaceae and Lachnospiraceae, and genus Ruminococcus, but reduce the abundance of genus Prevotella, Akkermansia and Oscillospira. Taken together, these results indicate that administration of EPS-09 can induce apoptosis of tumor cell, inhibit tumor angiogenesis, improve the immunity, regulate the intestinal microbiota composition of B16F10 melanoma bearing mice, and play positive roles in the antitumor activity against melanoma.

Nicotine plays a protective role in rats with induced viral pneumonia with polyinosinic-polycytidylic acid through α7nAChR

ObjectiveTo study the effect of nicotine in rat model of pneumonia induced by polyinosinic-polycytidylic acid [Poly (I:C)] and explore the underlying mechanism. MethodsTwenty-four healthy adult male Sprague-Dawley (SD) rats (200–250 g) were randomly divided into normal saline control group (NS group); Poly (I:C) group; nicotine group (NIC group); and α7 nicotinic acetylcholine receptor (α7nAChR) antagonist group (α-BGT group) (n = 6 each). Rats in the Poly (I: C), NIC, and α-BGT groups were administered 1.5 mg/mL 100 μL Poly (I:C) intranasally to establish pneumonia model. In α-BGT group, 1 μg/kg α-bungarotoxin (α-BGT) was intraperitoneally injected 45 min before intranasal Poly (I:C), and 400 μg/kg nicotine was intraperitoneally injected 15 min after α-BGT injection. The NIC group received an equal volume of NS in place of α-BGT while the other treatments were same. The Poly (I:C) group received equal volume of NS in place of nicotine while the other treatments were same as in NIC group. In the NS group, only NS was administered at all three time points. PaCO2, PaO2, and PaO2/FiO2 levels were determined 24 h after administration of Poly (I:C). After euthanization, rat lung tissues were extracted for pathological examination, and wet weight/dry weight (W/D ratio) was determined. Expression of tumor necrosis factor-α (TNF-α), interleukin (IL)-6, IL-1β, and interferon (IFN)-γ in lung tissue was determined by ELISA. q-PCR was used to detect nuclear factor kappa-B P65 (NF-κBP65). ResultsCompared with NS group, Poly (I:C) and α-BGT groups showed significantly increased W/D ratio, PaCO2, TNF-α, IL-6, IL-1β, and IFN-γ content, NF-κB P65 expression, and reduced PaO2 and PaO2/FiO2 (p < 0.05), along with obvious signs of pathological injury. Nicotine pre-treatment reduced W/D ratio, PaCO2, proinflammatory cytokines, NF-κBP65 expression, and increased PaO2 and PaO2/FiO2 levels. The above effects were negated in α-BGT group. ConclusionPre-administration of nicotine improved Poly (I:C)-induced pneumonia by activating the cholinergic anti-inflammatory pathway.

Comparison of the effect of moxibustion at "Zusanli" (ST36) on the polarization of synovial macrophages of knee joints in rats with knee osteoarthritis and rheumatoid arthritis.

To observe the similarities and differences of effects of moxibustion at "Zusanli" (ST36) on target tissues and macrophages polarization in knee osteoarthritis (KOA) and rheumatoid arthritis (RA) rats, and to summarize its efficacy and characteristics. Thirty rats were equally and randomly divided into control, KOA, RA, KOA+Moxi and RA+Moxi groups. The KOA model and RA model were induced by injection of sodium monoiodoacetate or Freund's complete adjuvant into the rats' knee joints, respectively. Rats of the KOA+Moxi and RA+Moxi groups received moxibustion stimulation at bilateral ST36 for 30 min, once a day for 21 days, beginning from the 7th day on after modeling. The contents of serum interleukin(IL)-1β and IL-10 were detected by ELISA. Histopathological changes (Markin score of the knee cartilage and synovial pathology score) of the knee joints were observed after HE staining. The polarization state of M1 and M2 macrophages in the synovial tissue of the knee joints was assessed by detecting the expression of CD86 and CD206 after immunofluorescence staining. Compared with the control group, the content of serum IL-1β, synovial pathology score, and synovial CD86 expression were significantly increased (P<0.01, P<0.05), while the content of serum IL-10 and synovial CD206 expression markedly decreased (P<0.01) in both KOA and RA groups；the Markin score was increased (P<0.01) in the KOA group. In comparison with the KOA group, the Markin score was obviously decreased (P<0.01), while the content of serum IL-10 and CD206 expression were apparently increased (P<0.01) in the KOA+Moxi group. No significant changes were found in the content of serum IL-1β, synovial pathology score and CD86 expression in the KOA+Moxi group relevant to the KOA group. In comparison with the RA group, the content of serum IL-1β, synovial pathology score, and CD86 expression were considerably decreased (P<0.01) in the RA+Moxi group. No marked differences were found in the serum IL-10 level, Markin score, and CD206 expression between RA+Moxi and RA model groups. The increased Markin score was significantly higher in the KOA group than in the RA group (P<0.01), but the increased synovial pathology score was significantly lower in the KOA group than in the RA group (P<0.01). Correspondingly, the effect of moxibustion at ST36 was significantly better in RA model than in KOA model in reducing serum IL-1β (P<0.05). Moxibustion at ST36 can effectively reduce cartilage injury of knee joint in rats with KOA and reduce synovial injury in rats with RA, which may be related with its effects in lowering IL-1β level in RA model by inhibiting the polarization of M1 macrophages, and up-regulating level of IL-10 in KOA model by promoting the polarization of M2 macrophages. However, the relevant mechanism needs to be further studied.

Study on the mechanism of electroacupuncture regulating macrophage polarization to improve ulcerative colitis in rats.

To investigate the effect of electroacupuncture(EA) at "Changbing Decoction" on alleviating ulcerative colitis (UC) and regulating the polarization of colonic macrophages in rats, so as to explore its mechanisms underlying improvement of UC. Twenty-six male SD rats were randomly divided into 4 groups：normal group(6 rats), model group(8 rats), EA group(6 rats), and western medication group(6 rats). The rat model of UC was established by using 5% dextran sulfate sodium (DSS) solution drinking water for 7 days, followed by drinking 1% DSS solution during treatment period. After 7-day model establishment, EA treatment(10 Hz/50 Hz, 20 min) was applied to "Zhongwan"(CV12), bilateral "Tianshu"(ST25) and "Shangjuxu"(ST37) for 3 d, and rats in the western medication group were given mesalazine suspension(200 mg/kg) by gavage for 3 d. The body weight, spleen weight and colon length of rats were measured. The disease activity index (DAI) score was evaluated. The morphological changes and inflammatory cell infiltration of colon were detected after HE staining and pathological scores were eva-luated. The contents of tumor necrosis factor α(TNF-α), interleukin(IL)-1β, IL-2 and IL-10 in serum were detected by ELISA. The protein expressions of M1 and M2 macrophage markers nitric oxide synthase (iNOS) and arginase 1(Arg1) were detected by fluorescence double staining and Western blot, respectively. Quantitative real-time PCR was used to detect iNOS and Arg1 mRNA expressions. Compared with the normal group, rats in the model group had increased pathological damage degree and inflammatory cell infiltration in the colon tissue, slowed-down body weight gain, decreased colon length, spleen weight, serum anti-inflammatory factors IL-2 and IL-10 contents, colonic Arg1/CD68 fluorescence positive expression, and Arg1 protein and mRNA expressions(P<0.01, P<0.05), as well as increased DAI scores, colon histopathological scores, contents of serum pro-inflammatory factors TNF-α and IL-1β, colonic iNOS/CD68 fluorescence positive expression, iNOS protein and mRNA expressions(P<0.01). Compared with the model group, the above indicators were significantly improved in rats of the EA group and the western medication group(P<0.01, P<0.05). EA of "Changbing Decoction" can improve UC of rats by regulating the polarization of colonic macrophages, inhibiting the generation of M1 macrophages and promoting the generation of M2 macrophages.

In vitro and in vivo antibacterial studies of volatile oil from Atractylodis Rhizoma against Staphylococcus pseudintermedius and multidrug resistant Staphylococcus pseudintermedius strains from canine pyoderma

Ethnopharmacological relevanceAtractylodis Rhizoma is extensively employed in Traditional Chinese Medicine for the treatment of skin and gastrointestinal ailments. Its active components have been proven to demonstrate numerous beneficial properties, including antibacterial, antiviral, anti-inflammatory, anti-tumor, and anti-ulcer activities. Furthermore, the volatile oil from Atractylodis Rhizoma (VOAR) has been reported to effectively inhibit and eradicate pathogens such as Staphylococcus aureus, Escherichia coli and Candida albicans. Of particular concern is Staphylococcus pseudintermedius, the predominant pathogen responsible for canine pyoderma, whose increasing antimicrobial resistance poses a serious public health threat. VOAR merits further investigation regarding its antibacterial potential against Staphylococcus pseudintermedius. Aim of the studyThe study aims to verify the in vitro antibacterial activity of VOAR against Staphylococcus pseudintermedius. And a superficial skin infection model in mice was established to assess the in vivo therapeutic effect of VOAR. Materials and methodsThirty strains of S. pseudintermedius were isolated from dogs with pyoderma, and the drug resistance was analyzed by disc diffusion method. The Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of VOAR were determined through the broth dilution method. The growth curve of bacteria in a culture medium containing VOAR was monitored using a UV spectrophotometer. Scanning electron microscopy was employed to observe the effects of VOAR on the microstructure of S. pseudintermedius. The impact of VOAR on the antibiotic resistance of S. pseudintermedius was assessed using the disc diffusion method. Twenty mice were randomly divided into four groups: the control group, the physiological saline group, the VOAR group, and the amikacin group. With the exception of the control group, the skin barrier of mice was disrupted by tap stripping, and the mice were subsequently inoculated with S. pseudintermedius to establish a superficial skin infection model. The modeled mice were treated with normal saline, VOAR, and amikacin for 5 days. Following the treatment period, the therapeutic effect of each group was evaluated based on the measures of body weight, skin symptoms, tissue bacterial load, tissue IL-6 content, and histopathological changes. ResultsThe MIC and MBC of VOAR against 30 clinical isolates of S. pseudintermedius were found to be 0.005425% and 0.016875%, respectively. VOAR could exhibit the ability to delay the entry of bacteria into the logarithmic growth phase, disrupt the bacterial structure, and enhance the antibacterial zone in conjunction with antibiotic drugs. In the superficial skin infection model mice, VOAR significantly reduced the scores for skin redness (P < 0.0001), scab formation (P < 0.0001), and wrinkles (P < 0.0001). Moreover, VOAR markedly reduced the bacterial load (P < 0.001) and IL-6 content (P < 0.0001) in the skin tissues of mice. Histopathological observations revealed that the full-layer skin structure in the VOAR group was more complete, with clearer skin layers, and showed significant improvement in inflammatory cell infiltration and fibroblast proliferation compared to other groups. ConclusionThe results demonstrate that VOAR effectively inhibits and eradicates Staphylococcus pseudintermedius in vitro while also enhancing the pathogen's sensitivity to antibiotics. Moreover, VOAR exhibits a pronounced therapeutic effect in the superficial skin infection model mice.

Puerarin alleviates hyperosmotic stress-induced oxidative stress, inflammation, apoptosis and barrier damage of human corneal epithelial cells by targeting SIRT1/NLRP3 signaling

The increase of tear osmolarity caused by excessive evaporation of tear phase is the main pathological mechanism of dry eye disease (DED). Puerarin, the major bioactive ingredient isolated from the root of the Pueraria lobata (Willd.) Ohwi, has been reported to improve ophthalmic diseases in clinic. However, the effect and the potential regulatory mechanism related to silent information regulator sirtuin 1 (SIRT1)/NOD-like receptor family pyrin domain containing 3 (NLRP3) signaling of puerarin in DED has not been evaluated. In this study, we aimed to explore the effect and mechanism of hyperosmotic stress (Hyp)-induced human corneal epithelial cell line (HCE-2). The viability of HCE-2 cells induced by Hyp with or without puerarin treatment was assessed by a CCK-8 assay. Results indicated that puerarin treatment enhanced cell viability, reduced reactive oxygen species (ROS) content, increased CAT and SOD activities, and elevated the ratio of GSH/GSSG in HCE-2 cells exposed to Hyp. Besides, TNF-α, IL-1β and IL-6 contents were decreased by puerarin. Additionally, puerarin inhibited Hyp-induced apoptosis and barrier disruption of HCE-2 cells. Moreover, molecular docking method suggested that puerarin bound to SIRT1, and upregulated SIRT1 and downregulated NLRP3 inflammasome proteins after puerarin treatment was observed. Furthermore, SIRT1 silencing alleviated the protective effects of puerarin on Hyp-induced HCE-2 cell damage. Collectively, puerarin attenuates Hyp-induced injury of HCE-2 cells by targeting regulating SIRT1/NLRP3 signaling.

Immune synergistic mechanism of recombinant plasmid adjuvant containing chicken IL-4 and IL-2 fusion genes on chicken coccidia live vaccine

The recombinant plasmid pCI-IL-4-IL-2-EGFP containing fusion genes of chicken IL-4 and IL-2 can be used as an adjuvant to enhance the anticoccidiosis effect of the chicken coccidia live vaccine. The chickens were divided into 3 groups: blank control group, vaccine+pCI-IL-4-IL-2-EGFP adjuvant coimmunization group, and vaccine-only group to investigate the immune synergy mechanism of recombinant plasmid adjuvant pCI-IL-4-IL-2-EGFP. The expressions of IL-2, IL-4, TNF-α, and IFN-γ in chicken sera and tissues were detected by ELISA and RT-qPCR, and the proliferation of T and B lymphocytes and antigen presenting cells (APC) in chicken immune organs and intestines were detected by acid alpha-naphthalase (ANAE) staining, methyl green pyronine (MGP) staining, and immunofluorescence (IF) staining, respectively. Results showed that the mRNA expression of IL-2, IL-4, IFN-γ and the number of activated T and B lymphocytes were significantly upregulated in the spleen and cecum tonsils of chickens in vaccine+pCI-IL-4-IL-2-EGFP group compared with the vaccine-only group on 7 d after vaccination (P < 0.05). Protein contents of IL-2, IL-4 and TNF-α in vaccine+pCI-IL-4-IL-2-EGFP group were significantly increased compared to vaccine-only group on 28 d of inoculation (P < 0.05). The number of T and B lymphocytes and APC in chickens of the vaccine+ pCI-IL-4-IL-2-EGFP group was significantly higher than that of the vaccine-only group in cecum tonsils, thymus and spleen after 14 and 28 d of inoculation (P < 0.05). All results revealed that pCI-IL-4-IL-2-EGFP adjuvant enhanced the immune response of chicken coccidia live vaccine by upregulating the expression of IL-2, IL-4, TNF-α, and IFN-γ and promoting the proliferation of T, B lymphocytes and APCs in chicken intestines and immune organ sites. Moreover, our study provides a theoretical basis for the clinical application of cytogenic plasmids as adjuvants.

Post-ischemic treatment of nalmefene hydrochloride attenuated lung ischemia-reperfusion injury in rats via the Sirt1/Nrf2/HO-1 pathway with inhibition of ferroptosis

Objective: To study the effect and mechanism of post-ischemic treatment of nalmefene in alleviating the lung ischemia-reperfusion injury by inhibiting ferroptosis through activation of the Sirt 1/Nrf 2/HO-1 axis. Methods: A total of 60 rats were randomly divided into six groups equally (n=10): the sham group, the model group(I/R), the nalmefene group, the nalmefene+EX527 group, the nalmefene+ML385 group, the nalmefene+Fe-citrate group (nalmefene+Fe group). The sham group without drug treatment was not treated with ischemia-reperfusion. The pulmonary ischemia-reperfusion model was established by occlusion of the left pulmonary hilum in the model group without drug treatment. After ischemic treatment, the nalmefene group was injected with nalmefene (15 μg/kg) via the tail vein at 5 minutes before reperfusion. The nalmefene+EX527 group, the nalmefene+ML385 group, and the nalmefene+Fe group were injected intraperitoneally with EX527 (5 mg/kg), ML385 (30 mg/kg), Fe-citrate(15 mg/kg), respectively, 2 h before moulding and then injected with nalmefene (15 μg/kg) via the tail vein at 5 minutes before reperfusion. All rats were sacrificed three hours after reperfusion, and the specimens from the upper lobe of the left lung tissue were preserved. The degree of lung tissue injury and the wet/dry weight ratio were assessed in each group of rats. Fe 2+, MDA, TNF-α, and IL-6 content, GSH activity and the expression levels of Sirt1, Nrf2, HO-1, ACSL4 and GPX4 were determined. Results: Compared with the sham group, the wet/dry weight ratio, lung tissue injury score, ACSL 4 expression level, Fe 2+, TNF-α, IL-6 and MDA content, Sirt 1, Nrf 2, HO-1 messenger RNA and protein expression levels were significantly increased (P<0.01), while GPX 4 expression level and GSH activity were significantly decreased in the model group (P<0.01). Compared with the model group, wet/dry weight ratio, lung tissue injury score, ACSL 4 expression level, Fe 2+, TNF-α, IL-6, and MDA content decreased significantly (P<0.01), Nrf 2, HO-1 messenger RNA and protein, GPX 4 expression, and GSH activity were significantly increased in the nalmefene group and the nalmefene+EX527 group (P<0.01). Sirt 1 messenger RNA and protein expression increased significantly in the nalmefene (P<0.01) and the nalmefene+EX527 groups (P>0.05). In the nalmefene+ML385 group, the wet/dry weight ratio, lung tissue injury score, TNF-α and IL-6 content were decreased significantly (P<0.01), while Sirt 1 messenger RNA and protein expression levels were significantly increased (P<0.01), but there were no significant changes in Nrf 2, HO-1 messenger RNA and protein expression levels, ACSL 4 and GPX 4 expression levels, Fe 2+, MDA content, and GSH activity (P>0.05). In the nalmefene+Fe group, wet/dry weight ratio, lung-injury score, TNF-α, IL-6, MDA content were decreased significantly (P<0.01), messenger RNA and protein expression levels of Sirt 1, Nrf 2, HO-1, and GSH activity were increased significantly (P<0.01), but there were no significant changes in Fe 2+content, ACSL 4 and GPX 4 expression levels (P>0.05). Compared with the nalmefene group, in the nalmefene+EX527 group, the nalmefene+ML385 group and the nalmefene+Fe group, wet/dry weight ratio, lung tissue damage score, ACSL 4 expression level, TNF-α, IL-6 and MDA content were significantly increased (P<0.01), the expression level of GPX 4 and GSH activity were significantly decreased (P<0.01). The expression levels of Sirt 1, Nrf 2, HO-1 messenger RNA and protein were significantly decreased in the nalmefene+EX527 group (P<0.01). The expression levels of Nrf 2, HO-1 messenger RNA and protein decreased significantly in the namemefene+ML385 group (P<0.01), but there was no significant change in Sirt 1 messenger RNA and protein expression level (P>0.05). Sirt 1, Nrf 2, HO-1 messenger RNA-protein expression levels did not change significantly in the nalmefene+Fe group (P>0.05). Conclusion: Post-ischemic treatment with nalmefene hydrochloride may alleviate pulmonary ischemia-reperfusion injury by inhibiting ferroptosis through activation of the Sirt 1/Nrf 2/HO-1 axis.

Role and anti-inflammatory mechanisms of acupuncture and moxibustion therapy on pain relief through NOX-ROS-NLRP3 pathway in CCI rats models.

To observe the effects of acupuncture and moxibustion therapy on pain relief in sciatica rats and to explore the mechanism of its anti-inflammatory effect. SPF grade 4-6-week-old Kunming rats were randomly divided into 5 groups including a blank group, sham-operated group, model group, acupuncture, and moxibustion (AnM) group, and positive group. A total of 10 rats were included in each group. The model group, the AnM group, and the positive group were prepared by ligating the left sciatic nerve. AnM group was used for acupuncture and moxibustion therapy intervention, and the positive group was rendered to quick-acting sciatica pills once a day for 7 days (3 courses of treatment). The blank group, sham-operated group, and model group were not treated. The changes in thermal and mechanical pain thresholds were observed before and after the operation, and the morphological changes of the dorsal horn of the spinal cord in the lumbosacral region of the rats in each group were observed by HE staining after the courses of treatment finished. The contents of IL-1β, IL-6, IL-18, and TNF-α were measured by ELISA and the expressions of NOX1, NOX2, NOX4, and NLRP3 genes were detected by RT-qPCR while the protein expressions of NOX1, NOX2, NOX4 and NLRP3 were analyzed by Western blotting. The AnM and positive group showed a significant increase in thermal and mechanical pain thresholds after treatment, while there was no significant change in the model group. As compared to the control group, the contents of IL- 1β, IL-6, IL-18, and TNF-α, as well as the relative expressions of NOX1, NOX2, NOX4, and NLRP3 genes were significantly increased in the model group (P < 0.05 or P < 0.01). As compared to the model group, the contents of IL-1β, IL-6, IL-18, and TNF-α, as well as the relative expressions of NOX1, NOX2, NOX4, and NLRP3 genes significantly decreased in the AnM and positive groups (P < 0.05 or P < 0.01). The pathological changes of inflammatory infiltration of tissue cells in the dorsal horn of the lumbosacral spinal cord were slowed in the AnM group. Acupuncture and moxibustion therapy have a positive effect on pain relief and anti-inflammatory effects in CCI sciatica rats, which may point to the regulation of NOX1, NOX2, NOX4, and NLRP3 expressions, and inhibition of ROS.

Compound Bacillus alleviates diarrhea by regulating gut microbes, metabolites, and inflammatory responses in pet cats

BackgroundPet cats frequently have diarrhea in their daily life. Bacillus has a protective role that has crucial beneficial functions on intestinal homeostasis. The aim of this research was to investigate the effects of the compound Bacillus on the prevention of diarrhea, microbiota and metabolism in pet cats. A total of 20 pet cats (1–2 years old, 3.91 ± 0.92 kg) were randomly divided into two groups and fed with a basal diet (Control group), or a basal diet supplemented with 3 × 109 CFU/kg compound Bacillus (Probiotics group). The experiment lasted 33 days.ResultsResults showed that the compound Bacillus significantly reduced the rate of soft stools and diarrhea in pet cats compared with the control group (P < 0.05, n = 10). Meanwhile, compared with the control group, the probiotics group significantly decreased the content of IL-1β and IL-6 and significantly increased IL-10 (P < 0.05, n = 6) in the serum. In addition, feeding probiotics significantly increased the abundance of p_Patescibacter and g_Plectosphaerella, decreased the abundance of p_Firmicutes, p_Gemmatimonadetes, g_Ruminococcaceae_UCG-005, g_Ascochytahe and g_Saccharomyces in the feces of the pet cats (P < 0.05, n = 6). And it also can significantly increase the content of total SCFAs, acetic acid and butyric acid in the feces (P < 0.05, n = 6). The fecal and serum metabolomics analyses revealed that most fecal and serum compounds were involved in metabolism, particularly in chemical structure transformation maps and amino acid metabolism. Also, eugenitol and methyl sulfate were the most significantly increased serum metabolites, and log2FC were 38.73 and 37.12, respectively. Pearson’s correlation analysis showed that changes in serum metabolism and fecal microbiota were closely related to immune factors. There was also a strong correlation between serum metabolites and microbiota composition.ConclusionsThe results of this research highlight the potential of the compound Bacillus as a dietary supplement to alleviate diarrhea in pet cats.

Dietary Supplementation of Mixed Organic Acids Improves Growth Performance, Immunity, and Antioxidant Capacity and Maintains the Intestinal Barrier of Ira Rabbits.

The aim of this study was to investigate the effects of mixed organic acids (MOAs) on growth performance, immunity, antioxidants, intestinal digestion, and barrier function in Ira rabbits. A total of 192 weaned male Ira rabbits at 35 days of age were randomly assigned to four groups with six replicates of eight rabbits each. The rabbits in the control group (CON) were fed a basal diet, and the antibiotic group (SAL) was fed a basal diet supplemented with 60 mg/kg salinomycin. The test groups were fed a basal diet supplemented with 1000 mg/kg and 2000 mg/kg MOAs (MOA1 and MOA2, respectively). The experiment lasted for 55 days. The results showed that the ADG of Ira rabbits in the SAL group and MOA1 group was higher than that in the CON group (p < 0.05). The serum IL-6 and liver MDA levels of Ira rabbits in the SAL group, MOA1 group, and MOA2 group were lower than those in the CON group (p < 0.05). In addition, sIgA levels in the jejunal mucosa of Ira rabbits in the SAL group and MOA1 group were increased compared with those in the CON group (p < 0.05). Compared with the CON group, the gene expression of IL-6 was decreased (p < 0.05) in the jejunal mucosa of Ira rabbits in the SAL, MOA1, and MOA2 groups, while the gene expression of IL-1β tended to decrease (p = 0.077) and the IL-10 content tended to increase (p = 0.062). Moreover, the gene expression of ZO-1 in the jejunal mucosa of Ira rabbits was elevated in the MOA1 group compared with the CON group (p < 0.05). In conclusion, dietary supplementation with MOAs can improve growth performance, enhance immune function and antioxidant capacity, and maintain the intestinal barrier in weaned Ira rabbits.

Dietary supplementation with American ginseng dietary fiber ameliorates intestinal mucosal barrier injury in immunosuppressed mice

Dietary fiber is an important component of healthy eating patterns. Although American ginseng (Panax quiquefolium L.) is often used to improve body immunity, less attention has been paid to American ginseng dietary fiber (AGDF) in this regard. The aim of this study was to investigate the protective effect of AGDF on intestinal mucosal barrier injury induced by cyclophosphamide (CTX) in immunosuppressed mice and the underlying mechanism. The results showed that AGDF could increase immune organ index and enhance immunity of immunosuppressed mice, such as DTH response, lymphocyte transformation and proliferation and NK effector cell activity. At the same time, AGDF could restore the proportion of CD4+/CD8+ T lymphocytes induced by CTX to different extents, and promote the secretion of immunoglobulin IgG. More importantly, AGDF could up-regulate the contents of TNF-α, IFN-γ, IL-6 and IL-1β in serum and intestine of immunosuppressed mice, promote the secretion of cytokines and regulate the homeostasis balance between Th1/Th2 cytokines. It also upregulated the expression of tight junction proteins occludin, claudin-1 and ZO-1, and alleviated the degree of CTX-induced damage to the intestinal mucosal barrier. Meanwhile, AGDF could reduce intestinal epithelial cell apoptosis by activating TLR4/NF-κB signaling pathway, improving the phosphorylation of JNK and p38 signaling molecules, which may be the potential mechanism by which AGDF ameliorates intestinal mucosal injury in immunosuppressed mice. Therefore, AGDF may be used as a dietary supplement to prevent and treat intestinal mucosal barrier injury caused by CTX.

Efficacy and mechanism of Wuzi Yanzong pill on the prevention and treatment of EAE

ObjectiveStudies have shown that Wuzi Yanzong Pill (WYP) can be used to treat neurological diseases, but its mechanisms for multiple sclerosis (MS) remain unclear. This study aims to determine the effect of WYP on MS in an animal model of experimental autoimmune encephalomyelitis (EAE), and explore its mechanism. To provide theoretical basis for the clinical treatment of MS with WYP. MethodsC57BL/6 female mice were randomly divided into Blank control, EAE control, low dose WYP, medium dose WYP, and high dose WYP groups. One week before model generation, the mice were gavaged with saline (50 mL/kg/d) in Blank control and EAE control groups. The treatment groups was gavaged with different doses of WYP solution (4, 8, or 16 g/kg/d respectively) Clinical scores were recorded daily. Sample collection was conducted on the 14th and 28th days, respectively The expressions of IL-10, IL-17, IL-12, TNF-α and IFN-γ in spleen were detected by ELISA. The expressions of ROCKII, P-MYPT1, TLR4, NF-κB/p65, MCP-1, CCR2 in spleen, brain and spinal cord were detected by Western Blot. The types of macrophages and the contents of intracellular IL-10 and IL-12 were detected by Flow Cytometry. The contents of TNF-α and TLR4 mRNA in the spleen were detected by RT-PCR. ResultsWYP treatment improved the clinical score of EAE mice in a significant dose-dependent manner, with the WYP high-dose group showed the most significant improvement in clinical score. Compared with the EAE control group, WYP high dose group had significantly lower levels of IL-17, IFN-γ, ROCKII, P-MYPT1, TLR4, NF-κB/p65, MCP-1, and CCR2 as well as TNF-α and TLR4 mRNA, but increased the number of M2 macrophages and IL-10. ConclusionWYP treatment relieves clinical symptoms in EAE mice, which may be related to regulate inflammatory pathway and inhibiting expressions of inflammatory cytokines.

Chonggu Granules Improve Cartilage Matrix Metabolism in Knee Osteoarthritis via the miR-148a-3p/Wnt/β-Catenin Pathway.

This study aims to explore the effect and underlying mechanism of Chonggu Granules (CGG) in knee osteoarthritis (KOA) in rats. A papain-induced KOA model was established in rats. The pathological alterations of extracellular matrix in rat cartilage tissues were observed through hematoxylin and eosin (H&E) staining, followed by Mankin score for quantitative scoring. The ultrastructure of cartilage extracellular matrix was examined under a transmission electron microscopy (TEM). ELISA was used to measure the levels of IL-6, TNF-α, and IL-1β in rat serum. Immunofluorescence was performed for assessing the levels of MMP-3, MMP-13, and Col2al in rat cartilage. Western blot was used to identify the protein expressions of wnt1, GSK-3β, β-catenin, and Aggrecan in rat cartilage. The mRNA relative expressions of miR-148a-3p, wnt1, β-catenin, and GSK-3β in rat cartilage were detected by RT-PCR. Luciferase reporter gene was used to detect the target genes of miR-148a-3p. CGG significantly improved articular cartilage tissue and extracellular matrix metabolism compared to the model group as indicated by H&E, Mankin score, and TEM data. Moreover, low, medium, and high doses of CGG reduced the levels of IL-6, TNF-α, IL-1β, MMP-3, and MMP-13 in serum to varying degrees but increased the levels of Col2al and Aggrecan. Mechanistically, CGG targeted wnt1 by increasing the expression of miR-148a-3p in a dose-dependent manner, thereby downregulating the mRNA and protein expressions of β-catenin in cartilage tissue and upregulating the mRNA and protein expressions of GSK-3β. CGG may control the miR-148a-3p/wnt/β-catenin signaling pathway to decrease the levels of its downstream target genes MMP-13 and MMP-3, increase the expressions of Col2al and Aggrecan, and downregulate the contents of inflammatory cytokines IL-6, TNF-α, and IL-1β, thereby improving the metabolism of cartilage extracellular matrix and alleviating the degeneration of articular cartilage in KOA.

Exposure to bromoxynil octanoate herbicide induces oxidative stress, inflammation, and apoptosis in testicular tissue via modulating NF-кB pathway.

Bromoxynil octanoate (BO) is a herbicide necessary for plant growth and production. However, it may cause damage to environment and humans. This study aimed to investigate the potential testicular toxicity of BO and its possible underlying mechanisms. Male Albino (Sprague Dawley) rats were administered BO in different doses (5, 10, 20, and 40mg/kg/BW; P.O.) daily for 21 days. Testicular function was evaluated by determining count and viability of epididymal sperm, and testosterone. In addition, the following parameters were assessed; MDA, NO, and H2O2 as oxidative stress markers; SOD, CAT, GPx, GST, and GSH as antioxidant markers; NF-ĸB-P65 and IL-18 as inflammatory markers; caspase-9 and caspase-3 as apoptotic markers; gene expression of NF-ĸB-P65, TNF-α, BAX, Bcl-2, and caspase-3; and histopathological examination of epididymis and testis sections. The results showed a significant (P<0.05) increase in MDA, NO, H2O2, IL-18, and caspase-9 content, NF-ĸB-P65, TNF-α, Bax, and Caspase-3 expression as compared to control. Furthermore, the count and viability of epididymal sperm, testosterone level, SOD, CAT, GPx, GST, and GSH content, and Bcl-2 expression showed a significant (P<0.05) decrease as compared to control. In conclusion BO-induced testicular damage by altering oxidation, inflammation, and apoptosis.