ABSTRACT
Objective To investigate the effects of arsenic trioxide (ATO) on the expression of autoan-tibody and interleukin (IL)-10 IL-12 in MRL/lpr mice. Methods MRL/lpr mice wereseparated into 3 different groups. The 3 groups received arsenic trioxide (ATO, 0.4 mg·kg~(-1)·d~(-1)), cyclophosphamide (CTX,50 mg/kg) and sodium chloride (NS, volume weight-determined) abdominal injection respee-tively. The treatment stopped 2 months later. Afterwards, the rates of CD3~+(T) cells, CD3~+CD4~+(Th) cells and the CD3~+CD4~+cells which produced IL-10 and IL-12 were detected using single-cell measurement of intr-acellular eytokines by flow cytometry after polyclonal stimulation with PMA and ionomycin for 4 hours in 5% CO_(2.)Serum levels of IL 10 and IL-12 were assessed using the Mouse cytokines ELISA Kit. One-way ANOVA LSD test and paires t test were used for statistical analysis.Results ①The level of anti-dsDNA antibody after treatment was 0.92±0.06, while it was 1.14±0.58 before treatment. So the ds-DNA antibody level was significantly decreased in ATO group (P<0.01), while it was dramatically increased in the NS groups (P<0.05) after the treatment;②ATO group had significantly less CD3~+ cells and CD3~+CD4~+ cells[(44±4)% and (20±4)%]compared withNS group [(59±5)%and(30±3)%](P<0.01).③The serum level of IL-12 in the ATO group was (84±12) pg/ml,while it was (103±13)pg/ml in the NS group (P=0.018).④The intracellular levels of IL-10 and IL-12 produced by CD3~+CD4~+ (Th) cells in the ATO group were ( 1.5±0.4)% and (2.43±0.42)%, which was significantly lower than those in the NS group respectively (2.5±0.5)% and (3.24±0.40)%(P<0.01). Conclusion Arsenic trioxide can reduce the production of anti-dsDNA antibody,inhibit the activation and proliferation of both T cells and Th subsets in the MRLApr mice, and hence decrease the serum levels of IL-12 and the levels of IL-10, IL-12 produced by Th cells.
ABSTRACT
Objective To investigate the effects of andrographolide on the expressions of Th1 cytokine [interferon (IFN)-γ] mRNA and Th2 cytokines [interleukin (ID-4, IL-10] mRNA of peripheral blood mononuclear cells (PBMCs) from patients with chronic hepatitis B (CHB) and its anti-hepatitis B virus (HBV) activity. Methods PBMCs from CHB patients were cultured with 10 mg/L andrographolide (experimental group) or 0.1% DMSO (control group). HepG2. 2. 15 cells were stimulated with andrographolide of different concentrations (experimental group) or adefovir (control group). The expressions of IFN-γ mRNA, IL-4 mRNA and IL-10 mRNA of PBMCs and the replication of HBV DNA in HepG2. 2. 15 cell line were detected by fluorescence quantitative PCR. Results The expression of IFN-γ mRNA in PBMCs cultured with 10 mg/L andrographolide for 16 h was higher than that in control group (Z=-2. 78, P=0. 05), and the expressions of IL-4 and IL-10mRNA in experimental group were lower than control group (Z= -3. 82, P<0. 01), while the ratio of IFN-γ/IL-4 mRNA was higher than control group (Z= - 3. 82, P<0. 01). Andrographolide with different concentrations had no effect on the replication of HBV DNA in HepG2. 2.15 cells ((=11. 88, P>0.05). However, adefovir had inhibitory effect on the replication of HBV DNA (t =15. 95,P< 0. 05). Conclusion Andrographolide can regulate the expressions of IFN-γ mRNA, IL-4 mRNA and IL-10 mRNA in PBMCs from CHB patients and improve Thl/Th2 balance, while it has no effect on the replication of HBV DNA.
ABSTRACT
Objective To investigate the effects of alpha-melanocyte-stimulating hormone (alpha-MSH) on the production of tumor necrosis factor-alpha (TNF-alpha) and intedeukin-10 (IL-10) by peri-pheral blood monohuclear cells (PBMCs) from patients with psoriasis vulgaris. Methods Heparinized peri-pheral blood was obtained from 20 patients with psoriasis vulgaris and 10 healthy human controls. PBMCs were isolated, cultured in complete medium, and stimulated with phytohemagglutinin (PHA) alone, the com-bination of PHA and various concentrations of alpha-MSH, or nothing. After another 48-hour culture, ELISA and real-time PCR were performed to measure the secretion levels of TNF-alpha and IL-10 in the super-natants of cultured PBMCs as well as the mRNA expression levels of TNF-alpha and IL-10 in PBMCs. Results The secretion level of TNF-alpha in the supematants of patient-derived PBMCs stimulated by nothing or PHA alone was significantly higher than that from normal control-derived PBMCs (329.87 ± 99.33 ng/L vs 116.95 ± 37.15 ng/L, 1756.01 ± 183.60 ng/L vs 1287.30 ± 152.36 ng/L, both P<0.01). alpha-MSH of all tested concentrations (10-13, 10-11, 10-7,mol/L) could inhibit the secretion of TNF-alpha by PBMCs com-pared with PHA alone (all P < 0.01), and the maximum effective concentration was 10-13 mol/L. On the con-Wary, a significant decrease was observed in the secretion level of IL-10 in the supematants of patient-derived PBMCs stimulated by nothing or PHA alone compared with normal control-derived PBMCs (P <0.05 or 0.01). Moreover, the secretion of IL-10 by PBMCs was promoted by alpha-MSH of all tested con-centrations (P < 0.01 or 0.05), with the maximum effective concentration being 10-13 mol/L (P < 0.01). The alpha-MSH of 10-13 mol/L down-regulated the mRNA expression of TNF-alpha (P < 0.001), but up-regnlated that of IL-10 (P < 0.001) in PHA-stimulated PBMCs from patients. Conclusion alpha-MSH can regulate the production of TNF-alpha and IL-10 by PHA-stimulated PBMCs from patients with psoriasis vulgaris.
ABSTRACT
Objective To investigate the effect of adenoviral-mediated interleukin-10 (IL-10) gene transfection on systemic inflammatory responses in a rat model of ventilator-induced lung injury. Methods Thirty adult male SD rats weighing 200-220 g were randomly divided into 5 groups (n=6 each): group Ⅰ control (group C); group Ⅱ adenovirus without IL-IO gene + 2 h mechanical ventilation (MV) (group B, ); group II1 adenoviruswithout IL-IO gene + 4 h MV (group B_2); group Ⅳ adenovirus with IL-10 gene + 2 h MV (group A_1) and group Ⅴ adenovirus with IL-10 gene + 4 h MV (group A_2). In group Ⅱ-Ⅴ the animals were mechanically ventilated (airway pressure = 25 cm H_2 O) and adenovirus was injected into right ventricle at 48 h before MV. Blood samples were taken from abdominal aorta at the end of 2 h or 4 h MV for determination of serum IL-8 and IL-10 concentrations. The animals were then killed. The pulmonary specimens were obtained for examination of ultrastructure with light and electron microscope. Results Mechanical ventilation significantly increased serum IL-8 and IL-IO concentrations in a duration-dependent manner in group Ⅱ-Ⅴ as compared with group C. Trans-pulmonary administration of adenoviraI-mediated interleukin-10 gene significantly decreased serum IL-8 concentration and increased serum IL-10 concentration and reduced ventilator-induced lung injury alter MV in group A_1 and A_2 as compared with group B_1 and B_2. Conclusion Adenoviml-madiated IL-10 gene transfection can attenuate the systemic inflammatory response and reduce ventilator-induced lung injury.