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Objective:To explore the effect of leptin on B cells in patients with systemic lupus erythematosus (SLE).Methods:Peripheral blood mononuclear cells (PBMCs) were isolated from SLE patients, and then CD19 + B cells were purified with magnetic bead sorting method. PBMCs or purified B cells were cultured with recombinant leptin at 0, 100, 250 ng/ml for 3 or 5 days. The frequencies of plasma cells, follicular helper T (Tfh) cells and peripheral helper T (Tph) cells, as well as activation markers (CD80, CD86) and leptin receptor and the proliferation of B cells were determined with flow cytometry. The concentrations of antibodies and cytokines were examined with enzyme-linked immunosorbnent assay (ELISA). Data were analyzed with t test and analysis of variance (ANOVA). Results:Increased levels of leptin were positively correlated with systematic lupus erythematosus disease activity index (SLEDAI) and the frequency of plasma cells in SLE patients. Leptin receptor could be detected on SLE B cells, and recombinant leptin elevated the levels of its receptor on CD19 + B cells [(7.8±1.3)% vs (6.1±0.9)%, t=3.36, P=0.006]. Leptin enhanced the expression of CD80 [(21±4)% vs (19±4)%, t=2.84, P=0.004] and CD86 [(22±4)% vs (19±4)%, t=4.92, P=0.004] on SLE B cells in vitro. It also promoted B cells to differentiate into plasma cells [(7.6±1.5)% vs (5.2±1.3)%, t=6.42, P=0.025]. There was no statistical significant difference of the effect of leptin on B cell proliferation. Leptin also increased the levels of antibodies [IgG: (62±3) ng/ml vs (45±4) ng/ml, t=7.75, P<0.001; IgM: (112±24) ng/ml vs (56±18) ng/ml, t=5.38, P<0.001] and inflammatory cytokines [IL-6: (24±5) pg/ml vs (20±5) pg/ml, t=4.09, P=0.002; TNF-α: (19.1±3.8) pg/ml vs (14.1±2.9) pg/ml, t=3.38, P=0.006; IL-10: (24±5) pg/ml vs (20±5) pg/ml, t=4.09, P=0.002] secreted by B cells. In addition, leptin significantly upregulated the frequencies of Tfh cells[(2.82±0.49)% vs (1.28±0.20)%, t=4.56, P=0.001] and Tph cells [(4.5±0.5)% vs (3.4±0.4)%, t=3.88, P=0.003]. Conclusion:Leptin could directly promote the activation, differentiation and secretory capacity of B cells by binding to its receptor, and also modulate B cell responses indirectly via enhancement of Tfh and Tph cells, which may be involved in the pathogenesis of SLE.
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Objective To investigate the effects of protein expressions and the urea transport activity of aldosterone on urea transporter A1 (UT-A1) and urea transporter A3 (UT-A3) in HEK293 cells and Xenopus laevis oocytes.Methods (1) Western Blot was used to investigate the protein expressions of UT-A1 and UT-A3.(2) Cell surface biotinylation was used to investigate the protein expressions of UT-A1 and UT-A3 on the cell surface of Xenopus laevis oocytes.(3) 14C-urea transport experiment was conducted to investigate the transport activity of UT-A1 and UT-A3 in Xenopus laevis oocytes.Results (1) Compared with UT-A1 or UT-A3 high expression groups,the total protein levels of UT-A 1 and UT-A3 were all significantly reduced in aldosterone treatment groups (all P ＜ 0.01).(2) Compared with UT-A1 or UT-A3 high expression groups,the levels of protein expression on cell surface were all significantly reduced in aldosterone groups (all P ＜ 0.01).(3) Compared with UT-A1 or UT-A3 high expression groups,14C-urea transport experiment results showed that aldosterone treatment groups had significantly reduced the urea transporter activity of UT-A1 (1 min:94.32±9.044vs 40.68±4.274,P＜0.01,n=6;3 min:165.0±4.7 vs 80.3±0.6,P＜0.01,n=6),and UT-A3 (1 min:204.6± 3.1 vs 176.7± 9.1,P＜0.05,n=6;3 min:371.4 ± 14.9 vs 318.8 ± 12.0,P＜0.05,n=6).Conclusion Aldosterone can directly down-regulate the protein expressions of UT-A1 and UT-A3 in both total protein and cell surface level,which reduces their urea transport activity.
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The pathogenic and therapeutic effect of gut microbiome is a hot topic in recent years .Many researchers gradually focused on the relationship between the eating disorder and the gut microbiome .The gradual in-depth studies show that gut microbiome can intervene in the host′s metabolic state by multiple approaches to regulate appetite and bring new inspiration for the treatment of dis -eases.In this article, we review all important findings in this field and discuss the mechanism how gut microbiome effects appetite regu -lation.
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Objective To evaluate the effect of methylene blue(MB)preconditioning on ischemi-a-reperfusion(I∕R)injury in isolated rat lungs. Methods Eighteen pathogen-free healthy male Sprague-Dawley rats, aged 3 months, weighing 240-320 g, were divided into 3 groups(n=6 each)using a ran-dom number table: sham operation group(group Sham), lung I∕R group(group I∕R)and methylene blue preconditioning group(group MB). A model of isolated lung I∕R injury was established in pentobarbi-tal sodium-anesthetized rats. MB 2 mg∕kg was intraperitoneally injected at 2 h before stopping perfusion in group MB. Isolated lungs were perfused for 20 min, followed by 45-min ischemia, and then reperfused for 60 min in I∕R and MB groups. At 60 min of reperfusion, the activity of lactic dehydrogenase(LDH)in the perfusate was detected, wet weight(W)and dry weight(D)was determined, W∕D ratio was calcu-lated, and the levels of malondialdehyde(MDA), ATP, reactive oxygen species(ROS)and superoxide dismutase(SOD)were measured in lung tissues. Mitochondria and cytoplasm were isolated from lung tis-sues for determination of mitochondrial membrane potential(MMP), degree of mitochondrial swelling and content of cytochrome C(Cyt c)in cytoplasm. Apoptotic cells in lung tissues were detected using TUNEL, and apoptotic index was calculated. Results Compared with group Sham, the activity of LDH in perfu-sate, W∕D ratio, levels of ROS, MDA and Cyt c in cytoplasm and apoptosis index were significantly in-creased, the degree of mitochondrial swelling was aggravated, and the content of ATP and MMP were de-creased in I∕R and MB groups, and the SOD activity was significantly decreased in group I∕R(P<005). Compared with group I∕R, the activity of LDH in perfusate, W∕D ratio, levels of ROS, MDA and Cyt c in cytoplasm and apoptosis index were significantly decreased, the degree of mitochondrial swelling was attenu-ated, and the activity of SOD, content of ATP and MMP were increased in group MB(P<005). Con-clusion Methylene blue preconditioning can reduce I∕R injury in isolated rat lungs, and the mechanism may be related to improving mitochondrial function and inhibiting cell apoptosis.
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Objective To observe the influence of Qingwen Baidu decoction (QBD) on serum procalcitonin (PCT) and C-reactive protein (CRP) levels in septic rats and study the mechanism of heat-clearing and detoxifying method for treatment of sepsis. Methods Fifty male Sprague-Dawley (SD) rats were randomly divided into control group (n=5), model group (n=25), and heat-clearing and detoxifying experimental group (experimental group, n=20). The septic model was reproduced by intra-peritoneal injection of lipopolysaccharide (LPS, 5 mg/kg). In control group, an equal volume of normal saline was given. After modeling for 2 hours, the heat-clearing and detoxifying experimental group received QBD the first time (composition of the decoction: Gypsum Fibrosum Recens 30 g, Rehmanniae Radix 10 g, Bubali Cornu 15 g, Coptidis Rhizoma 4 g, Gardeniae Fructus 5 g, Platycodonis Radix 5 g, Scutellariae Radix 5 g, Anemarrhenae Rhizoma 5 g, Paeoniae Radix Rubra 5 g, Scrophulariae Radix 5 g, Forsythiae Fructus 5 g, Glycyrrhizae Radix 5 g, Moutan Cortex 5 g, Lophatheri Herba 5 g) by gavage (0.01 mL/g); the rest administration time was 08:00 to 09:00, once a day. The rats in model group were given an equal volume of warm water by gavage. At different time points after modeling, the blood of 5 rats in control group, model group, and experimental group was collected from the abdominal aorta. The serum PCT and CRP levels were tested by the enzyme linked immunosorbent assay (ELISA), and the pathological changes in lung and intestinal tissue were observed under a light microscope. Results Compared with the control group, the PCT level of the model group after modeling for 2 hours was significantly increased (ng/L:332.32±22.85 vs. 70.46±3.18, P 0.05). The level of CRP in model group was lower than that of control group at 24 hours and 72 hours after modeling (μg/L:281.34±32.81, 237.84±41.42 vs. 350.09±56.67, P 0.05). The PCT level of experimental group was significantly lower than that of model group beginning from 48 hours after modeling (ng/L: 321.57±28.00 vs. 358.12±10.14, P < 0.05), and this situation continued until 72 hours after modeling (ng/L: 269.50±49.10 vs. 347.69±26.90, P <0.05). The CRP level of experimental group was significantly lower than that of model group beginning from 8 hours after modeling (μg/L:232.73±13.29 vs. 335.35±53.78, P<0.05), this statistical significant difference between the two groups persisted until 72 hours after modeling (μg/L:177.31±6.70 vs. 237.84±41.42, P<0.05). Compared to those in the model group, the lung tissue inflammatory cell infiltration, the intestinal mucosal inflammation and interstitial edema were milder in the experimental group. Conclusion Heat-clearing and detoxifying therapy can effectively reduce the serum PCT and CRP levels of septic rats induced by LPS, and it can alleviate the infiltration of inflammatory cells in lung tissues so as to play a role in protection of tissue organ.
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Objective To study the relationship between the serum selenium(Se) concentra- tion and glutathione peroxidase(GSH-PX) levels in red blood cell in patients with pregnancy induced hypertension(PIH).Methods The serum Se concentration was measured by atomic absorbability assay and the GSH-PX levels were measured by cellular chemiluminescence assay.49 cases of PIH and 34 normal pregnant women were measured for serum Se concentration and GSH-PX levels in red blood cell from April,1994 to December,1994.Results The serum Se concentration and GSH-PX levels in red blood cell is different betwen PIH and control group.Compared serum Se concentration and GSH-PX levels were significant lower in the PIH patients than in the control subjects (P