OBJECTIVES: To investigate the therapeutic effects of cimifugin on Crohn's disease (CD)-like colitis in mice and its possible mechanism. METHODS: Thirty adult male C57BL/6 mice were randomized equally into control group, 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced CD-like colitis model group, and cimifugin treatment (daily gavage at 12.5 mg/kg) group. The therapeutic effect of cimifugin was evaluated by observing changes in body weight, disease activity index (DAI) scores, colon length, histopathological inflammation scores, and inflammatory cytokine levels in the colonic mucosa. Intestinal barrier integrity in the mice was assessed using immunofluorescence assay and Western blotting for claudin-1 and ZO-1; T-helper (Th) cell subset ratios in the mesenteric lymph nodes were analyzed with flow cytometry. Network pharmacology, KEGG enrichment analysis and molecular docking were used to predict the targets of cimifugin and analyze the key pathways and cimifugin-MAPK protein interactions, which were validated by Western blotting in the mouse models. RESULTS: In mice with TNBS-induced colitis, cimifugin treatment significantly attenuated body weight loss and colon shortening, lowered DAI and histopathological scores, decreased IFN-γ and IL-17 levels, and increased IL-4 and IL-10 levels in the colonic mucosa. Cimifugin treatment also significantly improved TNBS-induced claudin-1 dislocation and reduction of goblet cells, upregulated claudin-1 and ZO-1 expressions, reduced Th1 and Th17 cell percentages, and increased Th2 and Treg cell percentages in the colonic mucosa of the mice. KEGG analysis suggested a possible connection between the effect of cimifugin and MAPK signaling, and molecular docking showed strong binding affinity between cimifugin and MAPK core proteins. Western blotting demonstrated significantly decreased phosphorylation levels of JNK, ERK, and p38 in the colonic mucosa of cimifugin-treated mouse models. CONCLUSIONS Cimifugin alleviates TNBS-induced CD-like colitis by repairing intestinal barrier damage and restoring Th1/Th2 and Th17/Treg balance via suppressing MAPK pathway activation.
Animals ; Mice, Inbred C57BL ; Male ; Mice ; Crohn Disease/immunology* ; Colitis/immunology* ; MAP Kinase Signaling System/drug effects* ; Trinitrobenzenesulfonic Acid ; T-Lymphocytes, Helper-Inducer/drug effects* ; Intestinal Mucosa ; Disease Models, Animal

OBJECTIVETo observe the change of Th immunological gene in renal transplant recipients after the treatment of cyclosporine (CsA) and tacrolimus (FK506).

METHODSThe peripheral blood lymphacytes just before and 24 hours after CsA and FK506 treatment were isolated. The total RNA of them were reverse-transcripted and examined by real-time quantity PCR array. The results were analyzed by bioinformatic methods.

RESULTSThe TLR4, CEBPB, IL4R, IL1R1,IL18R1,and IL1R2 genes were remarkably upregulated, whereas IL-2, CCL5, CD27, CCR5, CCR4, CD4, RPL13A, TGFB3, CD86, CCR3, STAT1, NFATC2IP, IL23A, IL15, IRF4, and TFCP2 were downregulated 24 hours after CsA treatment. The IL18, IL7, PTPRC, TNFSF4, SPP1, GFI1, TLR4, IL13RA1, TNF, INHBA, LAG3, IL13, IL1R1, SOCS5, IL10, YY1, TBX21, FASLG, IL18R1, and IL1R2 genes were remarkably upregulated, whereas IL-2, IL-3, IL-4, IL-6,CCR5, CD4, CD27, CD40LG, IL15, CCR3, CD86, CCR4, and IRF4 were obviously downregulated 24 hours after FK506 treatment.

CONCLUSIONCsA and FK506 exert their therapeutic effectiveness by regulating the expressions of a series of target genes.

Cyclosporine ; pharmacology ; Cytokines ; genetics ; metabolism ; Gene Expression Regulation ; Humans ; Kidney ; drug effects ; metabolism ; Kidney Transplantation ; Oligonucleotide Array Sequence Analysis ; T-Lymphocytes, Helper-Inducer ; drug effects ; metabolism ; Tacrolimus ; pharmacology

BACKGROUNDSurgical stress causes a helper T-cell type 2 (Th2)-dominant status and disturbs the Th1/Th2 cytokine balance. Anesthesia can suppress the stress response to surgery, therefore it may inhibit the imbalance in the Th1/Th2 ratio. In this study, we assessed if propofol anesthesia and sevoflurane anesthesia influence the Th1/Th2 cytokine balance, and which anesthesia method better attenuates this ratio.

METHODSTwenty-eight patients with an American Society of Anesthesiologists (ASA) physical status of I undergoing laparoscopic cholecystectomy were selected. They were randomly allocated into two groups of 14. Group 1 received propofol anesthesia by a target-controlled-infusion (TCI) pump and group 2 received sevoflurane anesthesia. Non-invasive blood pressure, heart rate, and end-expiration CO2 partial pressure were monitored during anesthesia. The depth of anesthesia was measured using the bispectral index (BIS), and maintained between 50 and 60. During surgery we adjusted the doses of propofol and sevoflurane according to the BIS. Samples of peripheral blood were taken before the induction of anesthesia (T1), after the induction of anesthesia (T2), at the beginning of surgery (T3), at the end of surgery (T4) and on the first day after surgery (D1). Blood samples were analyzed to give the Th1/Th2 ratio and plasma level of cortisol.

RESULTSNon-invasive blood pressure, heart rate and end-expiration CO2 partial pressure were not notably different in the two groups. At T4, the percentage of T1 cells was higher in group 1 and had statistical significance (P < 0.05). The percentage of T2 cells was not significantly different in the two groups. At T4, the difference in the Th1/Th2 ratio was significantly different. At T3, T4, and D1, the plasma level of cortisol was lower in group 1 (P < 0.05).

CONCLUSIONCompared with sevoflurane, propofol can preferably promote Th cells to differentiate into Th1 cells and inhibit surgical stress. Propofol may therefore be immunoprotective for such patients.

Adult ; Cell Differentiation ; drug effects ; Female ; Flow Cytometry ; Humans ; Hydrocortisone ; blood ; Male ; Methyl Ethers ; pharmacology ; therapeutic use ; Middle Aged ; Propofol ; pharmacology ; therapeutic use ; T-Lymphocytes, Helper-Inducer ; cytology ; drug effects ; Th1 Cells ; cytology ; drug effects ; Th2 Cells ; cytology ; drug effects

OBJECTIVETo study the effects of huai qi huang, a traditional Chinese medicine, on cytokines Th1, Th2 and Th17 levels and alveolar macrophage phagocytosis in asthmatic rats sensitized by ovalbumin (OVA).

METHODSForty male Sprague-Dawley rats were randomly divided into five groups: normal control, untreated asthma, budesonide-treated, huai qi huang-treated and budesonide+huai qi huang-treated asthma (n=8 each). Asthma was induced by OVA sensitization and challenge. The levels of IL-4, IFN-γ and IL-17 in plasma and bronchoalveolar lavage fluid (BALF) were measured using ELISA. The phagocytosis of alveolar macrophages which were isolated and purified from BALF was evaluated by the colorimetric assay.

RESULTSThe levels of IL-4 and IL-17 increased, in contrast, the IFN-γ level decreased in plasma and BALF in the untreated asthma group compared with those in the normal control group. The IFN-γ level in the huai qi huang-treated asthma group was higher than that in the untreated asthma group. The IFN-γ level increased and the IL-17 level decreased more significantly in the budesonide+huai qi huang-treated asthma group when compared with the budesonide and huai qi huang alone treatment groups. The phagocytosis of alveolar macrophages in the untreated asthma group was lower than that in the normal control group. Huai qi huang alone or combined with budesonide increased the phagocytosis of alveolar macrophages compared with the normal control, untreated asthma and budesonid-treated asthma groups. The levels of IFN-γ in plasma and BALF were positively correlated with the phagocytosis of alveolar macrophages.

CONCLUSIONSThe levels of IL-4 and IL-17 increase and the IFN-γ level decreases in plasma and BALF, and the phagocytosis of alveolar macrophages decreases in asthmatic rats. Huai qi huang treatment may increase the IFN-γ expression in plasma and BALF and the phagocytosis of alveolar macrophages in asthmatic rats. There is a synergistic effect between huai qi huang and glucocorticoids.

Animals ; Asthma ; drug therapy ; immunology ; Cytokines ; biosynthesis ; Macrophages, Alveolar ; drug effects ; immunology ; Male ; Medicine, Chinese Traditional ; Phagocytosis ; drug effects ; Rats ; Rats, Sprague-Dawley ; T-Lymphocytes, Helper-Inducer ; immunology ; Th1 Cells ; immunology ; Th17 Cells ; immunology ; Th2 Cells ; immunology

OBJECTIVETo investigate the enhancing effect of Chinese medicine-Xuebijing injection on lipopolysaccharide (LPS) -induced apoptosis of CD4+ CD25+ regulatory T cells (Tregs) and polarization of helper T cells (Th).

METHODSCD4+ CD25+ Tregs collected from rat spleen in vitro by immunomagnetic beads assay were divided into the control group, anti-CD3/CD28 group, anti-CD3/CD28 + LPS group, anti-CD3/CD28 + "Xuebijing injection" group and anti-CD3/CD28 + LPS + "Xuebijing injection" group. Tregs apoptosis rate and expression of winged helix transcription factor (Foxp3) in Tregs were detected by flow cytometry on 3rd post culture day. CD4+ CD25- T cells were co-cultured with CD4+ CD25- Tregs (1:1) for 68 hours with canavalin A stimulation. Interferon gamma (gamma-IFN), interleukin (IL)-4 and IL-17 in supernatants, which respectively was secreted by Th1, Th2 and Th17, were measured by ELISA.

RESULTSTregs apoptosis rate of anti-CD3/CD28 + LPS + "Xuebijing injection" group (45.1 +/- 2.7%) was significantly higher than that of anti-CD3/CD28 + LPS group (29.4 +/- 1.6%, P < 0.01). Meanwhile, Foxp3 expressions in Tregs in above 2 groups were 95 +/- 9 and 140 +/- 18 respectively, showing statistically significant difference between them (P < 0.01). Gamma-IFN levels secreted in anti-CD3/CD28 + LPS + "Xuebijing injection" group were significantly higher than those in anti-CD3/CD28 + LPS group (P < 0.01), while IL-4 levels had an opposite tendency compared with gamma-IFN (P < 0.05), resulting in a marked increase in the ra- tio of gamma-IFN/IL-4 in anti-CD3/CD28 + LPS + "Xuebijing injection" group (P < 0.01). In anti-CD3/ CD28 + "Xuebijing injection" group, IL-17 secretion levels were significantly decreased compared with anti-CD3/CD28 group (P < 0.05).

CONCLUSIONSActivation of CD4+ CD25+ Tregs induced by LPS may mediate Th1 shift to Th2 response. "Xuebijing injection" can effectively regulate immune function of T cells, increase the LPS-induced apoptosis of CD4+ CD25+ Tregs as well as enhance the polarization of Th2 to Th1, thereby abating the suppressive state of cell-mediated immunity.

Animals ; Apoptosis ; Drugs, Chinese Herbal ; pharmacology ; Endotoxins ; Lipopolysaccharides ; Male ; Rats ; Rats, Wistar ; T-Lymphocytes, Helper-Inducer ; drug effects ; immunology ; T-Lymphocytes, Regulatory ; drug effects ; immunology

OBJECTIVETo investigate the effects of L-arginine in different doses on the serum levels of helper T lymphocyte1 (Th1)/Th2 cytokines in severely burned patients.

METHODSTwenty-nine severely burned patients, with total burn surface area from 50% to 80%TBSA, hospitalized within 20 hours after burn, were randomly divided into control group (10 cases, fed with 5% glucose saline 500 mL), L-arginine 200 mg group (10 cases, fed with 5% glucose saline 500 mL + 200 mg/kg L-arginine), L-arginine 400 mg group (9 cases, fed with 5% glucose saline 500 mL + 400 mg/kg L-arginine). All patients received enteral feeding through nasointestinal tube, started within 22 hours after burn. Fasting venous blood of all patients was harvested on post burn day (PBD) 1 (before enteral feeding), 3, 5, and 7 to determine serum contents of TNF-alpha, IL-1beta, TGF-beta(1) and IL-4 by radio-immunity method and enzyme-linked immunosorbent assay.

RESULTSSerum contents of TNF-alpha and IL-1beta of patients in all groups increased rapidly after burn, and although contents of TNF-alpha (318 +/- 57) ng/mL and IL-1beta (218 +/- 47) pg/mL of patients in L-arginine 200 mg group peaked on PBD 5, they were still significantly lower than those of patients in control group [(389 +/- 34) ng/mL, (272 +/- 40) pg/mL, P < 0.05], but they decreased on PBD 7. Serum contents of TNF-alpha and IL-1beta in L-arginine 400 mg group were close to those of control group (P > 0.05). Serum contents of TGF-beta(1) and IL-4 of patients in each group increased slowly after burn, and content of TGF-beta(1) (110 +/- 16) pg/mL of patients in L-arginine 200 mg group was significantly higher than that of patients in control group [(83 +/- 20) pg/mL, P < 0.05] on PBD 5. There was no statistical significant difference between L-arginine 400 mg group and control group in respect of serum content of TGF-beta(1) (P > 0.05).

CONCLUSIONSCompared with the dosage of 400 mg/kg L-arginine, the 200 mg/kg dose is more effective in reducing the release of Th1 cytokines and increasing Th2 cytokines production, hence maintaining Th1/Th2 cytokine ratio to produce better immune opsonization during the infection phase of severe burn.

Adolescent ; Adult ; Arginine ; administration & dosage ; therapeutic use ; Burns ; blood ; drug therapy ; Female ; Humans ; Interleukin-1beta ; blood ; Male ; Middle Aged ; T-Lymphocytes, Helper-Inducer ; metabolism ; Th1 Cells ; drug effects ; metabolism ; Th2 Cells ; drug effects ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; Young Adult

OBJECTIVETo study the principle of clearing Fei (), cooling blood, and detoxification as well as nourishing yin and moisening Fei (abbr. as CCD-NM) in regulating the levels of peripheral T-lymphocyte subsets Th and Tc cells to explore its mechanism for lowering the incidence of infection in patients with systemic lupus erythematosus (SLE).

METHODSSixty SLE patients without complicated infection were assigned to the treatment group and the control group, 30 in each group. The control group was treated with Western medicine alone, while the treatment group was treated with the same program of Western medicine, but additionally administered with either Langchuang No.1 (I) or 2 (II), serial concerted Chinese recipes, applied respectively in patients in the active stage or in the resting stage. The total time of treatment for both groups was 1 year. Further, a healthy control group was set up with 20 healthy subjects. The expressions of Th1, Th2, and Tc1 and Tc2 cells in peripheral blood were detected and compared with those in the healthy control group.

RESULTS(1) As compared with the healthy control group, ratios of Th1/Th2 and Tc1/Tc2 in SLE patients, whether complicated with infection or not, were significantly lower (P<0.05 or P<0.01). (2) Comparison between patients with complications and those uncomplicated with infection showed that the two ratios and Th1 expression were lower and Tc2 was higher in the former than those in the latter (all P<0.05). (3) Ratios of Th1/Th2 and Tc1/Tc2 increased after treatment in patients of both the treatment group and the control group (P<0.05 and P<0.01), but the changes in the treatment group were more significant (P<0.05).

CONCLUSIONThe principle of CCD-NM could regulate the Th and Tc subsets toward equilibrium in SLE patients, which might be one of the mechanisms of action for alleviating complicated infection.

Adolescent ; Adult ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Humans ; Lupus Erythematosus, Systemic ; drug therapy ; immunology ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Phytotherapy ; T-Lymphocyte Subsets ; drug effects ; T-Lymphocytes, Helper-Inducer ; drug effects

OBJECTIVETo investigate the therapeutic mechanism of three-step sequential method (TSSM) on patients with corticosteroid-dependent asthma (SDA).

METHODSForty patients with SDA were randomly assigned according to the randomizing number table to two groups equally, the treated group treated with three-step sequential recipes plus inhalation of Pulmicort Turbuhaler 200 microg, twice a day, and the control group treated with Pulmicort Turbuhaler alone. The therapeutic course for both groups was 12 - 14 weeks. Changes of the symptom score of asthma, the corticosteroid dosage used and the lung function were observed and the positive expression rate of IFN-gamma and IL-4 in peripheral CD4+ T cells were determined by flow cytometry before and after treatment.

RESULTSThere was significant difference in the asthma symptom score, the oral corticosteroid dosage and the lung function between the treated group and the control group after treatment (P < 0.01). The expression rate of Th2 reduced, the ratio of Th1/Th2 increased significantly after treatment in both groups (P < 0.01, P < 0.05), but the changes were more remarkable in the treated group than those in the control group, showing significant difference between them (P < 0.01), while the expression rate of Th1 had no obvious change after treatment with no significant difference shown between the two groups (P > 0.05).

CONCLUSIONTSSM can regulate imbalance of Th1/Th2, inhibit generation of inflammatory cytokines, decrease airway hyper-response, and therefore improve the pulmonary function, alleviate the asthmatic symptoms and reduce the patients' dependence on corticosteroid.

Adult ; Asthma ; drug therapy ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Female ; Glucocorticoids ; adverse effects ; therapeutic use ; Humans ; Male ; Middle Aged ; Phytotherapy ; methods ; Substance-Related Disorders ; drug therapy ; etiology ; T-Lymphocyte Subsets ; drug effects ; T-Lymphocytes, Helper-Inducer ; drug effects

OBJECTIVETo study the pathogenesis of IgA nephropathy (IgAN) and the effect of Yiqi Zishen Granule (YZG) on the helper T-lymphocyte subsets Th1/Th2 in IgAN patients.

METHODSIntracellular cytokines secreted by Th1/Th2 were measured using flow cytometry in 24 healthy subjects and 31 IgAN patients treated by YZG before and after treatment.

RESULTSLevel of interferon-gamma (IFN-gamma) was lower (P < 0.05) and levels of interleukin 4 (IL-4) and interleukin 10 (IL-10) were higher (P < 0.01) in the IgAN patients than those in the healthy controls, showing imbalance of Th1/Th2 in the IgAN patients. After YZG treatment, the level of IFN-gamma increased and IL-10 level decreased (P < 0.01).

CONCLUSIONImbalance of Th1/Th2 exists in IgAN patients. YZG has a regulatory effect on imbalance of Th1/Th2 in IgAN patients.

Adult ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glomerulonephritis, IGA ; drug therapy ; immunology ; Humans ; Interferon-gamma ; blood ; Interleukin-10 ; blood ; Male ; Phytotherapy ; Powders ; T-Lymphocytes, Helper-Inducer ; drug effects

Adjuvants, Immunologic ; pharmacology ; Asthma ; blood ; Cells, Cultured ; Cytokines ; metabolism ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; GATA3 Transcription Factor ; genetics ; metabolism ; Humans ; Interferon-gamma ; metabolism ; Interleukin-13 ; metabolism ; Interleukin-4 ; metabolism ; Interleukin-5 ; metabolism ; Leukocytes, Mononuclear ; cytology ; drug effects ; metabolism ; Male ; Oligodeoxyribonucleotides ; pharmacology ; Pulmonary Disease, Chronic Obstructive ; blood ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Statistics as Topic ; T-Box Domain Proteins ; T-Lymphocytes, Helper-Inducer ; cytology ; drug effects ; metabolism ; Th1 Cells ; cytology ; drug effects ; metabolism ; Th2 Cells ; cytology ; drug effects ; metabolism ; Transcription Factors ; genetics ; metabolism