Myricetin alleviates ovalbumin-induced allergic rhinitis in mice by regulating Th1/Th2 balance

Objective:To evaluate the effect of myricetin on ovalbumin(OVA)-induced allergic rhinitis in mice.Methods:Mice were sensitized and challenged using OVA(5%,500 mL)intraperitoneally and intranasally,respectively,on an alternative day for 14 days,followed by administration of myricetin(50,100,and 200 mg/kg)till day 21.Nasal symptoms,biochemical parameters,protein expressions,and histopathology were observed.Results:OVA-induced increased nasal symptoms including rubbing,sneezing,and discharge were significantly reduced by myricetin(100and 200 mg/kg)(P<0.05).Myricetin also protected against histamine challenge and attenuated elevated serum immunoglobulin E(IgE;total and OVA-specific),total IgG1,andβ-hexosaminidase levels,as well as leukotriene C4 and interleukins levels in nasal lavage fluid(P<0.05).Western blot analysis showed that myricetin significantly upregulated the protein expression of T-box expressed in T cells,while downregulating the protein expression of GATA binding protein 3,NF-κB,and IκB-α(P<0.05).Additionally,OVA-induced histopathological abberations in the nasal mucosa was markedly ameliorated by myricetin treatment(P<0.05).Conclusions:Myricetin exerts anti-allergic effects against OVAinduced allergic rhinitis via regulating Th1/Th2 balance.

Effect of glycyrrhetinic acid on Th1/Th2 balance in cough variant asthma mice

Objective:To evaluate the therapeutic effect of Glycyrrhetinic Acid on cough variant asthma(CVA)mice and to investigate the possible mechanism in reducing lung inflammation.Methods:48 young female Balb/c mice were divided into Control,CVA,Prednisone Acetate,Glycyrrhetinic Acid high-dose,Glycyrrhetinic Acid middle-dose and Glycyrrhetinic Acid lowdose groups randomly,with 8 mice in each group.The CVA mice model was established by ovalbumin(OVA)sensitization and OVA challenge,the animal asthma behavior was observed after drug administration,and the index of the lung of mice were recorded.The level of OVAsIgE in the bronchoalveolar lavage fluid(BALF)was tested by ELISA.The pathological changes of the lung tissue were observed by Hematoxylin and Eosin(H&E)staining.The protein expressions of T-bet,IFN-γ,Gata3,IL-4 and IL-13 in the lung tissue were determined by Western blot.Results:Compared with the CVA group,the index of lung of mice,the OVA-sIgE level in BALF and expression levels of Th2-related factor in the lung tissue of mice in Prednisone Acetate and Glycyrrhetinic Acid groups were significantly decreased(P<0.05 or P<0.01),the infiltration of inflammatory cells in the lung tissue was reduced,while expressions of Th1-related factor in the lung tissue was significantly increased(P<0.05 or P<0.01).Conclusion:Glycyrrhetinic acid has therapeutic effect on CVA mice,the underlying mechanism of Glycyrrhetinic acid alleviating lung impairment and airway inflammation may be associated with mediating the Th1/Th2 imbalance in the lung tissue.

All-transRetinoic Acid Regulates Th1/Th2 Balance in CD4+T cells When GATA-3 is Deficient

The essential effect of vitamin A on immune function occurs through various mechanisms including direct effect on ThloTh2 balance modulation. However, it is unclear whether or not vitamin A can regulate Thl-Th2 balance under a strong Thl-polarizing condition. Therefore, the purpose of our study was to examine the effect of vitamin A metabolite allotrans retinoic acid （ATRA） on ThloTh2 differentiation in CD4~ T cells under GATA-3 deficiency, which can induce Thl-polarizing condition. In the present study, GATA-3 deficiency T cells were induced by siRNA and checked by real-time quantitative PCR and western blot. GATA-3 deficiency CD4＋ T cells and normal CD4＋ T were treated for 48 h with or without ATRA.

Effect of Th1/Th2 balance on serum prostaglandin,COX-2,AVP and OT in primary dysmenorrhea rats with cold coagulation and blood stasis syndrome

Objective:Explore the role of Th1/Th2 balance in the pathogenesis of primary dysmenorrhea through study the effect of Th1/Th2 balance drift on serum prostaglandin F_(2α),prostaglandin,6-keto-prostaglandin F_(1α),TXB2,COX-2,AVP and OT in Primary Dysmenorrhea Rats with cold coagulation and blood stasis syndrome.Method:The rat model of primary dysmenorrhea with cold coagulation and blood stasis syndrome was established.On the basis of this model,Th1 shift model and Th2 shift model were established.The rats were divided into blank group,model group,Th1 shifting group and Th2 shifting group.The writhing reaction of rats in each group was compared,and the contents of PGF_(2α),PGE_(2),6-keto-PGF_(1α),TXB2,COX-2,AVP and OT in serum were detected.Results:Writhing latency:There was no significant difference between Th1 and Th2 shifting groups(P>0.05),Th2 shifting group was shorter than Th1 shifting group(P<0.05).Writhing times in 30 minutes:Compared with the model group,the writhing times of Th1 shifting group decreased significantly(P<0.01),while Th2 shifting group increased significantly(P<0.01).Compared with the blank group,the contents of PGF_(2α),TXB2,COX-2,AVP and OT were increased(P<0.05,P<0.01),and the contents of 6-keto-PGF_(1α)and PGE_(2)were decreased(P<0.01).Compared with the model group,the contents of PGF_(2α),TXB2,COX-2 and AVP in Th1 shifting group were decreased(P<0.05,P<0.01),and the contents of 6-keto-PGF_(1α)and PGE_(2)were increased(P<0.01),while the contents of PGF_(2α),TXB2,COX-2,AVP and OT in Th2 shifting group were increased(P<0.01),and the contents of 6-keto-PGF_(1α)and PGE_(2)were decreased(P<0.01).Conclusion:The intervention of Th1/Th2 cell balance on the expression of PGF_(2α),PGE_(2),6-keto-PGF_(1α),TXB2,COX-2,AVP and OT may be one of the pathogenesis of PD with cold coagulation and blood stasis syndrome.

Scutellarin alleviates complete freund’s adjuvant-induced rheumatoid arthritis in mice by regulating the Keap1/Nrf2/HO-1 pathway

Scutellarin(SCU)is a herbal flavonoid glucuronide with multiple pharmacological activities,including antioxidant,anti-inflammation,vascular relaxation,anti-platelet,and myocardial protection.However,the effect of SCU on complete Freund’s adjuvant(CFA)-induced rheumatoid arthritis(RA)had not been studied.In this study,we investigated the beneficial effects of SCU in the CFA-induced RA mice model and the anti-arthritic activity was evaluated by paw edema.Enzyme-linked immunosorbent assay(ELISA)was carried out to evaluate the plasma levels of immunoglobulin(Ig)G,IgE,tumor necrosis factor(TNF)-α,interleukin(IL)-1β,IL-6,receptor activator of nuclear factor-κB ligand(RANKL),and osteoprotegerin(OPG).Histological slides were prepared from the harvested paws of mice to determine the pathological changes in the joints.The proportions of T helper type 1(Th1)and T helper type 2(Th2)cells of CD4+T lymphocyte subsets were analyzed by flow cytometry.The expression of Kelch-like ECHassociated protein 1(Keap1),nuclear factor erythroid 2-related factor 2(Nrf2),and heme oxygenase-1(HO-1)was analyzed using real-time quantitative PCR(RT-qPCR)and western blotting assays.The present study demonstrated that SCU prevented CFA-induced RA,and inhibited the expression of inflammation factors,IgG,IgE,TNF-α,IL-1β,and IL-6.While SCU also reduced the RANKL level,it increased OPG expression in RA mice.The Th1/Th2 ratio was significantly lower in mice treated with SCU.Additionally,HO-1 expression was reduced while the expression of Keap1 and Nrf2 was elevated following SCU treatment.Results provide preliminary evidence to employ SCU in arthritis treatment which might be related to the regulation of Th1/Th2 balance and the Keap1/Nrf2/HO-1 pathway.

Potential immunomodulatory role of sesamin in combating immune dysregulation associated with COVID-19

The ongoing outbreak of novel coronavirus disease 2019(COVID-19),caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),has caused an unprecedented global health crisis.Development of a cure for this devastating disease is currently at full speed,with several vaccines against COVID-19 already authorized and administered.Currently,demand for these vaccines far exceeds supply.As such,boosting immunity represents a viable route to halt the rapid spread of SARS-CoV-2 and limit fatalities until vaccines become more readily and widely available.The use of phytochemicals appears to be a promising panacea.Sesamin,a lignan isolated from Sesamum indicum seeds,is known for its potent pharmacological properties,and is therefore hypothesized as a potential candidate in the therapeutic regimen against COVID-19.Herein,we highlight the confirmed therapeutic anti-inflammatory and immune-modulatory potential of sesamin against myriads of respiratory disorders,and tentatively suggest that sesamin may exert similar potent effects against COVID-19.Precisely,we speculate that sesamin may help alleviate COVID-19 via restoring Th1/Th2 balance and preventing inflammation and cytokine storm development.Additionally,we further support the promising role of sesamin against COVID-19 by underscoring the direct evidence,which suggests that sesamin may demonstrate promising inhibitory potential against three important SARS-CoV-2 targets,namely main protease,spike protein,and angiotensin-converting enzyme 2 receptor.Although preliminary,there is ample evidence to propose sesamin as a potential phytotherapeutic and prophylactic candidate against COVID-19.Further in vitro,in vivo,and preclinical studies are required to further substantiate the role of sesamin in the prevention and/or treatment of COVID-19.

Study on prevention effect of Zishen Yutai pill combined with progesterone for threatened abortion in rats

Objective:To observe preventive and therapeutic effects and the mechanism of actions on Zishen Yutai pill combined with progesterone on threatened abortion in rats.Methods:After pregnancy,50 SPF female SD rats were selected and divided into control group,model group,progesterone group,Zishen Yutai pill group,and progesterone plus Zishen Yutai pill group(combination group),with 10 rats in each group.The rats of control group and abortion model group were lavaged with 2 mL/kg normal saline on pregnancy day 1 for continuous 10 days.Rats in Zishen Yutai pill group were given 1.575 g/kg/d of Zishen Yutai pill for intragastric administration for continuous 10 days.Rats in progesterone group were given intramuscular injection treatment of 0.1 mL/d progestin,continuous for 10 days.Rats in combination group were given injection therapy of aqueous solution of Zishen Yutai pill for continuous 10 days,and other treatments were the same as previous two groups.Abortion model were established then and live births,numbers of abortion and average rate of abortion were compared between the five groups.Peripheral blood was collected to detect the estradiol(E2) and progestational hormone(P),and obtain ratio of Th1/Th2 cytokines(IL-2,INF-γ,IL-4,IL-10).Results:Significant more live births of rats were found in the control group compared with other four groups(P<0.05).The numbers of live births of the rats in abortion model group were significantly less than that of progesterone group,Zishen Yutai pill group and joint group(P<0.05).The numbers of live births of rats in joint group were significant more than that of progesterone group and Zishen Yutai pill group.The serum E2 level of P of rats in the control group,progesterone group,Zishen Yutai pill group and joint group were significant higher than that of abortion model group(P<0.05).Serum levels of E2 and P of rats in the control group,progesterone group,Zishen Yutai pill group and joint group were not significant different(P>0.05) but these levels in the control group and Zishen Yutai pill group were significant lower than that of progesterone group and joint group(P<0.05).The maternal-fetal interface IL-4/IL-2,IL-10 / IL-2 of model group were balanced deviating to Th1,while the IL-4/IL-2,IL-10/IL-2 of progesterone group,Zishen Yutai pill group and joint group were balanced deviating to Th2,and joint group' deviation was better than progesterone group and Zishen Yutai pill group(P<0.05).Conclusions:Zishen Yutai pill combined with progesterone has a significant control effect for threatened abortion,which can obviously increase contents of maternal serums E2 and P,and regulate the Thl/Th2 balance with a remarkable effect.

Hydrolyzed seawater pearl tablet modulates the immunity via attenuating Th1/Th2 imbalance in an immunosuppressed mouse model

OBJECTIVE: To investigate whether Hydrolyzed Seawater Pearl tablet(HSPT) could modulate the Th1/Th2 imbalance in an immunosuppressed mouse model with Th1 to Th2 shift induced by Cyclosporine A(Cs A) which can be used in the clinical treatment of Th2 to Th1 shift diseases, and explore the possible mechanism for the adjuvant therapeutic efficacy of HSPT on recurrent respiratory infections(RRI) and acquired immune deficiency syndrome(AIDS).METHODS: The mice were randomly divided into six groups of five animals each, namely normal group, model group, lentinan polysaccharide tablet(LPT) group and three HPST treated groups. HPST treated groups were administered with HPST(0.51,1.02, 2.04 g/kg) via intragastric gavage(i.g) for 30 consecutive days. LPT used as reference drug for positive control, LPT group was administered with LPT(8.2 mg/kg) for 30 consecutive days. Normal group and model group were received distilled water. The animals in model group, LPT group and HPST treated groups were injected intraperitoneally with Cs A(50 mg/kg) to establish the immunosuppressed mice model with Th1 to Th2 shift on the20 th, 22 nd and 24 th day, one hour after the administration of the respective treatment. Animals were sacrificed one hour after the last administration to collect blood and splenic tissue. The proportion of T cells including CD8+ and CD4+ T cells, Th1 and Th2 in peripheral blood of experimental mice were measured by flow cytometric. The protein level in serum and m RNA level in splenic tissue of experimental mice for interleukin(IL)-2, IL-12, interferon-γ(IFN-γ), IL-4, IL-6, IL-10 and IL-13 were measured by enzyme linked immunosorbent assay and fluorescence quantitative polymerase chain reaction respectively.RESULTS: HSPT elevated the proportion of T cells including both CD8+ and CD4+ T cells, in which the proportion of Th1 and Th2 cells increased, while the ratio of Th1/Th2 cells decreased in peripheral blood of the immunosuppressed mouse model with Th1 to Th2 shift induced by Cs A. Furthermore,HSPT elevated both protein and m RNA level of Th1-type cytokines IL-2 and IFN-γ, while had no significant effect on protein and m RNA level of Th1-type cytokine IL-12 and Th2-type cytokines IL-4, IL-6, IL-10, IL-13 in mouse model.CONCLUSION: Our findings suggest that HSPT can increase proportion of T cells including both CD8+and CD4+T cells and induce Th2 to Th1 shift in both cells and cytokines, which probably was the mechanism to account for the adjuvant therapeutic efficacy of HSPT on RRI and AIDS.

Wang-Bi tablet, a patented Chinese medicine, maintains the balance of Th1/Th2 in mice with collagen-induced arthritis

OBJECTIVE: To investigate the pharmacological mechanism of Wang-Bi tablets(WBTs), a Chinese patented medicine, in rheumatoid arthritis(RA) using mice with collagen-induced arthritis(CIA).METHODS: A mouse model of CIA was induced using bovine type Ⅱ collagen. WBT treatment was administered and efficacy was evaluated. The levels of interferon-γ(IFN-γ), interleukin-2(IL-2), and interleukin-4(IL-4) were examined using an enzyme-linked immunosorbent assay, and the proportions of Th1 and Th2 were detected using flow cytometry. T-bet and GATA-binding protein 3(GATA3)expression were demonstrated using Western blot analysis.RESULTS: Paw swelling and the arthritis index decreased significantly following WBT treatment. Histopathological analysis revealed markedly alleviated damage to synovium tissue in the WBT and methotrexate treatment groups. WBT regulated the production of IFN-γ, IL-2, and IL-4 and modulated Th1 and Th2 cell populations, which might have been induced by the attenuation of Th1 and Th2 cell differentiation through a decrease in the expression of T-bet and an increase in the expression of GATA3 in the synovial tissue in CIA mice.CONCLUSION: These results indicate that WBT may produce a therapeutic effect on CIA through maintaining the balance of Th1/Th2 cells, which could result in a decrease in the autoinflammatory disorder observed in RA.

Common and small molecules as the ultimate regulatory and effector mediators of antigen-specific transplantation reactions

In spite of intensive research, the molecular basis of allograft and xenograft rejection still remains not fully understood. The acute rejection of an allograft is associated with the intragraft Th1 cytokine response, while tolerance of an allograft or xenograft rejection is accompanied by a higher production of the Th2 cytokines interleukin(IL)-4 and IL-10. Nevertheless, these cytokines are not the final regulatory and effector molecules mediating transplantation reactions. Data indicate that the functioning of common molecules with enzymatic activities, such are inducible nitric oxide synthase(i NOS), arginase, heme oxygenase-1(HO-1) or indoleamine-2,3-dioxygenase(IDO), the bioavailability of their substrates(L-arginine, tryptophan, heme) and the cytotoxic and regulatory actions of their small gaseous products(NO, CO) can be the ultimate mechanisms responsible for effector or regulatory reactions. Using models of transplantation immunity and tolerance we show that T cell receptor-mediated recognition of allogeneic or xenogeneic antigens as well as the balance between immunity/tolerance induces distinct cytokine production profiles. The ratio between Th1 and Th2 cytokines efficiently regulates the expression of genes for common enzymes, such as i NOS, arginase, HO-1 and IDO. These enzymes may compete for substrates, such as L-arginine or tryptophan, and the final product of their activity are small molecules(NO, CO) displaying effector or regulatory functions of the immune system. Thus, it is suggested that in spite of the high immunological specificity of transplatation reaction, the ultimate players in regulatory and effector functions could be small and common molecules.

Effect of acitretin on the T helper cells of psoriasis vulgaris

Objective To investigate the effects of acitretin on T helper cell(Th)1/Th2 balance and Th17 cells in psoriasis vulgaris(PV)patients.Methods A total of 13 men and 17 women with PV were investigated.10 mg of acitretin was administered twice a day for 8 weeks for intervention therapy.Serum levels of interferon-gamma(IFN-γ),interleukin(IL)-4 and IL-17 were measured by enzyme-linked immunosorbent assay.T,Th1,Th2 and Th17 cells in skin biopsies were counted with double-labeled immunofluorescence.Psoriasis Area and Severity Index(PASI)score was calculated before and 8 weeks after treatment.Results Before treatment PV patients had higher serum levels of IFN-γ and IL-17,and increased T,Th1 and Th17 cells in skin biopsies.After treatment,both serum levels of IFN-γ and IL-17,and T,Th1 and Th17 cells infiltrating in PV skin decreased significantly.Th1/Th2 balance was restored to normal.However,their IL-4 and Th2 cells showed no significant change throughout the therapy.Conclusion Acitretin exerts influence on dermal Th1/Th2 balance and Th17 cell infiltration,so does it on production of systematic inflammatory cytokines IFN-γ and IL-17 in PV patients.However,Th2 cells and its derivative cytokine—IL-4 are not affected.

针灸对溃疡性结肠炎大鼠Th1/Th2免疫平衡的影响

目的:探讨针灸对溃疡性结肠炎(UC)大鼠Th 1/Th 2免疫平衡的影响。方法:SD大鼠随机分为模型组、隔药灸组、电针组,每组10只,并另选10只正常大鼠作为对照组。采用免疫学方法并加局部刺激建立大鼠UC模型。隔药灸组、电针组选取"天枢"(双)和"气海"进行隔药灸或电针治疗,连续治疗14d。采用定量PCR法检测结肠组织中干扰素(INF)-γ、白介素(IL)-12、IL-4和IL-10mRNA表达水平;采用酶联免疫吸附法检测结肠组织中INF-γ、IL-12、IL-4和IL-10的含量;采用流式细胞术分析结肠组织细胞中CD4^+INF-γ^+/CD 4^+IL-4^+的比值。结果:与对照组比较,模型组大鼠结肠组织中INF-γ、IL-12mRNA和含量显著升高(P<0.05),IL-4、IL-10mRNA和含量显著下降(P<0.05);隔药灸组和电针组大鼠结肠组织INF-γ、IL-12mRNA和含量显著低于模型组(P<0.05),IL-4、IL-10mRNA和含量显著高于模型组(P<0.05)。模型组结肠组织中CD 4^+INF-γ^+/CD 4^+IL-4^+的比值显著高于对照组(P<0.05);与模型组比较,隔药灸组和电针组CD4^+INF-γ^+/CD 4^+IL-4^+比值显著下降(P<0.05)。隔药灸组和电针组的各项指标比较差异均无统计学意义(P>0.05)。结论:隔药灸和电针均能显著改善UC模型大鼠炎性反应,通过下调结肠组织中INF-γ和IL-12水平,上调IL-4和IL-10水平从而保持Th 1/Th 2细胞间平衡,进而改善免疫功能。

Protective effects of catalpol and rhein in murine experimental autoimmune encephalomyelitis via regulation of T helper (Th)1, Th2,Th17, and regulatory T cell responses

OBJECTIVE:To examine the effects of catalpol and rhein on pro-and anti-inflammatory responses in C57 BL/6 mice with experimental autoimmune encephalomyelitis(EAE),a model of multiple sclerosis.METHODS:Female C57 BL/6 mice were randomly divided into four groups(n=30):(a)normal salinecontrol,(b)EAE control,(c)EAE+prednisone acetate(PA,6 mg/kg),and(d)EAE+catalpol(40 mg/kg)and rhein(5 mg/kg).EAE was induced by injection of myelin oligodendrocyte glycoprotein 35-55 plus pertussis toxin.Treatments were orally administered daily for 40 d.Disease progression and neurological function were assessed using a semi-quantitative scale of tail and limb paralysis.Brains and spinal cords were collected on Days 6,20,and 40 and assessed for histopathological changes by hematoxylin and eosin staining.Production of interleukin(IL)-2,IL-4,IL-10,and IL-17 A protein was measured by enzyme-linked immunosorbent assay.Expression of the T helper(Th)1-,Th2-,Th17-,and regulatory T cell(Treg)-specific transcription factors T-bet,GATA3,ROR-γt,and Foxp3,respectively,were analyzed by quantitative reverse-transcription polymerase chain reaction and western blot analysis.RESULTS:Combination treatment with catalpol and rhein significantly alleviated the clinical disability and neurological dysfunction of mice with EAE.Catalpol and rhein treatment also reduced the infiltration of pro-inflammatory T cells into pathological lesions;significantly increased the expression of the anti-inflammatory factors GATA3,Foxp3,IL-4,and IL-10;and significantly decreased the expression of the pro-inflammatory factors T-bet,ROR-γt,IL-2,and IL-17 A.CONCLUSION:Catalpol and rhein reduced the neurological disabilities of mice with EAE,at least in part by rebalancing the pro-and anti-inflammatory environment in the brains and spinal cords.

Notch signaling pathway mediates the immunomodulatory mechanism of Yangfei Huoxue decoction alleviating bleomycin-induced pulmonary fibrosis in rats

OBJECTIVE: To investigate the immunomodulatory mechanism by which Yangfei Huoxue decoction(YHD) alleviates bleomycin(BLM)-induced pulmonary fibrosis(PF) in rats.METHODS: Rats were randomly divided into two time-point groups(day 14 and 28), and each timepoint group comprised the following six subgroups: control, BLM, dexamethasone(DXM), YHD high dose(YHD-H), YHD middle dose(YHD-M), and YHD low dose(YHD-L). Haematoxylin and eosin and Masson staining, flow cytometry, enzymelinked immunosorbent assay, Western blotting and UPLC-QT of analyses were examined.RESULTS: The results showed that YHD reduced the degree of alveolar inflammation and fibrosis;downregulated the expression of CD28, CD80,CD86, Delta-like 1, Notch2, and Notch3;and upregulated the proportions of Th1/Th2 and Tc1/Tc2. The seven components of YHD were detected.CONCLUSION: The current study indicates that YHD mainly functions by regulating the immune system and that the molecular mechanism may be related to the regulation of the Notch signaling pathway.

Effect of Xinfeng capsule on pulmonary function in a adjuvant arthritis rat model

OBjECTIVE:To observe the relationship between reduced pulmonary function and regulatory T cells(Tregs)and helper T cells(Th)1/Th2 drift in a rat model of adjuvant arthritis(AA),and to study the impact of Xinfeng capsule(XFC)on pulmonary function and investigate the mechanism of action.METHODS:Forty rats were randomly divided into normal control group(NC),model control group(MC),Tripterygium glycosides tablet group(TPT),and XFC group,with 10 in each.Except for the NCgroup,AA was induced in all rats by intracutaneous injection of 0.1 mL Freund's complete adjuvant in the right paw.On the 19th day after modeling,the NC and MC groups were given physiological saline(0.9%),while the TPT and XFC groups were given TPT(10 mg/kg)and XFC(2.4 g/kg),once daily,respectively.Thirty days after administration,changes in paw swelling,arthritis index(AI),pulmonary function,levels of serumγ-interferon(IFN-γ)and interleukin(IL)-4,Tregs in peripheral blood,and IFN-γ,IL-4,Forkhead box transcription factor 3(FoxP3)in lung tissue were observed by enzyme-linked immunosorbent assay,flow cytometry,polymerase chain reaction,and western blot.RESULTS:Compared with the NC group,paw swelling,AI,IFN-γ,and Th1/Th2 were increased,and pulmonary function parameters,IL-4,FoxP3 were decreased significantly in the MC group(P<0.05 or P<0.01).Pulmonary function parameters,Treg,IL-4,FoxP3(and mRNA)were higher,and paw swelling,AI,and IFN-γ(and mRNA)were lower in the XFC group than those in the MC group.The XFC group was also much better than theTPT group in improving pulmonary function,FoxP3 mRNA,IFN-γ,IL-4,Th1/Th2,and IL-10(P<0.05 or P<0.01).CONCLUSION:Xinfeng capsule can improve pulmonary function by regulating the levels of Tregs,inhibiting the activation of Th1 to Th2 cells,inducing drift,maintaining cell immune suppression,correcting the imbalance of Th1/Th2,and reducing inflammatory mediators.

Effect of chiropractic manipulation on disrupted epithelium barrier and its mechanism of specialized pro-resolving mediators in a spleen-deficiency murine model

OBJECTIVE: To investigate the influence of spleen deficiency on the epithelial barrier of jejunum and lungs in a rat model of spleen-deficiency and the effect and potential specialized pro-resolving mediators (SPMs) mechanism of chiropractic manipulation. METHODS: Three-week-old male Sprague-Dawley rats were divided randomly into normal control group (n = 6), spleen-deficiency group (n = 5) and chiropractic group (n = 6). Spleen-deficiency model was induced in spleen-deficiency group and chiropractic group. Moreover, chiropractic manipulation was performed in chiropractic group. Four weeks later, systemic Th1/Th2 balance was evaluated by the ratio of plasma interferon (IFN)-γ/interleukin (IL)-4 levels by enzyme-linked immunosorbent assay (ELISA). Epithelial barrier integrity were assessed by the observation of morphological changes by hematoxylin-eosin staining and zonula occludens (ZO)-1 gene expressions by quantitative real time polymerase chain reaction in jejunum and lungs. Plasma resolvin D1 (RvD1) and lipoxin A4 (LXA4) levels were measures by ELISA for endogenous SPMs production. The levels of docosahexaenoic acid (DHA) and arachidonic acid (AA) in jejunum and lungs were also measured by HPLC-MS/ MS. RESULTS: Comparing with normal control group, spleen-deficiency group showed disrupted mucosa in jejunum, inflammatory condition in lungs, significantly decreased ratio of plasma IFN-γ/IL-4 levels and lower expressions of ZO-1 mRNA in both jejunum and lung tissues. Comparing with spleen-deficiency group, chiropractic group had less disrupted mucosa in jejunum and inflammatory condition in lungs, significantly increased systemic ratio of IFN-γ/IL-4 and expressions of ZO-1 mRNA in both jejunum and lung tissues. Chiropractic group had significantly enhanced plasma levels of RvD1 and LXA4, but had no significantly higher levels of DHA and AA in jejunum and lungs when comparing with spleen-deficiency group. CONCLUSION: Spleen deficiency caused systemic Th1/Th2 imbalance towards Th2 polarization and epithelial barrier disruption in jejunum and lungs.Chiropractic manipulation helped enhance endogenous SPMs production, which might be one of the action mechanism of chiropractic manipulation on the improvement of epithelial barrier disruption.

Effects of appendectomy and oral tolerance on dextran sulfate sodium colitis

AIM： To evaluate the concomitant effects of appendec- tomy and oral tolerance on colitis.METHODS： Delayed-type hypersensitivity （DTH） was investigated at a 7-d interval after ovalbumin （OVA） ad- ministration and immunization under normal and colitis conditions in appendectomized or sham-operated mice. Pathological scores for the colon were graded after in- gestion of colon-extracted protein （CEP） and induction of dextran sulfate sodium （DSS） colitis in appendecto- mized or sham-operated mice. Thereafter, Thl and Th2 in Peyer＇s patches and spleen lymphocytes were de- tected in CEP-treated and bovine serum albumin （BSA）-treated control mice.RESULTS： In appendectomized mice, DTH was not inhibited at day 7 after OVA administration and at the initial phase of DSS colitis, whereas it was inhibited at day 14 and day 21. However, in sham-operated mice, it was inhibited during the whole procedure and the onset of DSS colitis. The protective role of CEP against DSScolitis was present in sham-operated mice, with pre- dominant improvement of colonic pathological changes, while vanished in the appendectomized mice. A shift from Thl to Th2 in Peyer＇s patches resulted from a de- crease of Thl cells with the ingestion of CEP. Compared with BSA in the sham-operated group, no predominant changes were observed in the appendectomized mice.CONCLUSION： Appendectomy interferes with the pro- tective role of CEP in DSS colitis via a shift from Th2 to Thl during oral tolerance induction.