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Objective To identify the relationship between nephritis activity, autophagy and inflammation in patients with SLE. Methods Western blot analysis was used to detect the expression of microtubule-associated protein 1 light chain 3 (LC3) and P62 in peripheral blood mononuclear cells (PBMCs) of SLE patients with lupus nephritis and non-lupus nephritis patients. Tumor necrosis factor α (TNF-α) and interferon γ (IFN-γ) in the serum of SLE patients were determined by ELISA. The correlation between LC3II/LC3I ratio and SLE disease activity score (SLEDAI), urinary protein, TNF-α and IFN-γ levels was analyzed by Pearson method. Results The expression of LC3 was increased and P62 was decreased in SLE patients. TNF-α and IFN-γ were increased in the serum of SLE patients. LC3II/LC3I ratio was positively correlated with SLEDAI (r=0.4560), 24 hour urine protein (r=0.3753), IFN-γ (r=0.5685), but had no correlation with TNF-α (r=0.04 683). Conclusion Autophagy is found in PBMCs of SLE, and the autophagy is correlated with renal damage and inflammation in patients with lupus nephritis.
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Humans , Tumor Necrosis Factor-alpha/metabolism , Leukocytes, Mononuclear/metabolism , Autophagy-Related Proteins/metabolism , Lupus Nephritis/urine , Kidney , Interferon-gamma/metabolism , Inflammation/metabolism , Lupus Erythematosus, Systemic/metabolismABSTRACT
Objective:To investigate the effect of irisin on pyroptosis in rats with ventilator-induced lung injury.Methods:Thirty-six healthy clean-grade male Sprague-Dawley rats, weighing 200-250 g, aged 6-8 weeks, were divided into 3 groups ( n=12 each) using a random number table method: control group (group C), ventilator-induced lung injury group (group V) and ventilator-induced lung injury plus irisin group (group V+ I). In group V+ I, irisin 1 μg/kg was injected via the tail vein before mechanical ventilation.The animals were mechanically ventilated (tidal volume of 40 ml/kg, respiratory rate 60 breaths/min, inspiratory/expiratory ratio 1∶2, positive end expiratory pressure 0 and inspired oxygen fraction ratio 21%.Blood samples were then taken from the femoral artery for blood gas analysis, and PaO 2 was recorded.Bronchoalveolar lavage fluid (BALF) was collected, the total protein concentrations in BALF were measured, and the concentrations of BALF and serum interleukin-1β (IL-1β) and IL-18 were measure by enzyme-linked immunosorbent assay.The lung tissues were obtained for determination of the pathological changes after HE staining which were scored, wet to dry weight (W/D) ratio, expression of pyroptosis-related proteins N-terminal gasdermin D (GSDMD-N) and caspase-1 protein and mRNA (by Western blot or using real-time polymerase chain reaction). Results:Compared with group C, the lung injury score and W/D ratio were significantly increased, PaO 2 and OI were decreased, the total protein concentrations in BALF, concentrations of IL-1β and IL-18 in BALF and serum were increased, and the expression of caspase-1 and GSDMD-N protein and mRNA was up-regulated in group V ( P<0.01). Compared with group V, the lung injury score and W/D ratio were significantly decreased, PaO 2 and OI were increased, the total protein concentrations in BALF, concentrations of serum IL-1β and IL-18 in BALF and serum were decreased, and the expression of caspase-1 and GSDMD-N protein and mRNA was down-regulated in group V+ I ( P<0.01). Conclusion:The mechanism by which irisin reduces ventilator-induced lung injury is probably related to inhibiting pyroptosis in rats.
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Objective:To investigate the value of α-synuclein (α-syn) concentration in cerebrospinal fluid (CSF) in predicting postoperative delirium (POD).Methods:One thousand patients underwent elective surgery with combined epidural-spinal anesthesia in our hospital from January 2018 to September 2020 were selected.The epidural puncture was performed at L 3, 4 interspace, and 2 ml of CSF was collected after the needle reaching the subarachnoid space.The concentrations of α-syn, β-amyloid (Aβ)40, Aβ42, total tau protein (T-tau), and phosphorylated tau protein (P-tau) in CSF were determined by enzyme-linked immunosorbent assay.The concentrations of α-syn in CSF and occurrence of POD in patients of different ages were recorded.Patients were divided into POD group and non-POD group according to whether POD occurred, and frequency matching (1∶1) was performed based on five matching variables of age, ASA physical status, education level, duration of operation, and intraoperative blood loss. Results:Eight hundred and forty-one patients were finally included in the study, and the incidence of POD was 15.0%. There were 126 cases in POD group and 126 cases in non-POD group after matching. The concentrations of α-syn in CSF and incidence of POD were gradually increased with age ( P<0.05). Compared with non-POD group, the concentrations of α-syn, T-tau and P-tau in CSF were significantly increased, the concentrations of Aβ40 and Aβ42 were decreased, Aβ40/P-tau, Aβ42/P-tau, Aβ42/Aβ40 and P-tau/T-tau were decreased in POD group ( P<0.05). After confounding factors were corrected by logistic regression analysis, increased concentrations of α-syn, p-tau, and T-tau in CSF were risk factors for POD ( P<0.05). Increased concentrations of Aβ40 and Aβ42 in CSF and increased Aβ40/P-tau and Aβ42/P-tau were protective factors for POD ( P<0.05). Multiple linear regression analysis showed that the concentration of α-syn in CSF was negatively correlated with Aβ40 and Aβ42 concentrations and positively correlated with P-tau and T-tau concentrations ( P<0.05). The area under the receiver operating characteristic curve of concentrations of α-syn in CSF predicting POD was 0.895, Youden index was 0.664, sensitivity was 80.00%, and specificity was 86.36% ( P<0.001). Conclusion:The concentration of α-syn in CSF is related to the occurrence of POD, and it provides higher accuracy in predicting POD.
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Objective:To evaluate the effect of electroacupuncture (EA) preconditioning on expression of cortical Ubc9 during cerebral ischemia-reperfusion (I/R) in rats.Methods:A total of 80 healthy clean-grade male Sprague-Dawley rats, aged 8-12 weeks, weighing 200-250 g, were divided into 4 groups ( n=20 each) using a random number table method: sham operation group (group S), group I/R, EA preconditioning group (group E) and sham EA group (group SE). Blood vessels were only exposed, without occlusion in group S. In the other three groups, the cerebral I/R model was established by middle cerebral artery occlusion using suture-occluded method, and the suture was removed after 2-h occlusion to restore the perfusion in anesthetized rats.EA was performed at 5 days before establishing the model in group E and group SE.Baihui acupoints were stimulated with an electric stimulator (2/12 Hz disperse-dense waves, intensity 1 mA) for 30 min once a day for 5 consecutive days, and the model was established at 24 h after the last stimulation.EA was performed at the points 1 cm lateral to the acupoints of Baihui, and the other operating parameters were the same as those previously described in group E. Neurological deficit scores (NDSs) were evaluated at 24 and 48 h of reperfusion.Then the rats were sacrificed, and tissues in the ischemic penumbra of cerebral cortex were obtained for determination of cell apoptosis (by TUNEL) and expression of Ubc9 and conjugated small ubiquitin-like modifier (SUMO) 2/3 (by Western blot). The apoptosis rate was calculated. Results:Compared with group S, NDSs at 24 and 48 h of reperfusion and apoptosis rate were significantly increased, and the expression of Ubc9 and conjugated SUMO2/3 was up-regulated in the other three groups ( P<0.05). Compared with group I/R and group SE, NDSs at 24 and 48 h of reperfusion and apoptosis rate were significantly decreased, and the expression of Ubc9 and conjugated SUMO2/3 was up-regulated in group E( P<0.05). Conclusion:The mechanism by which EA preconditioning reduces cerebral I/R injury is related to up-regulating Ubc9 expression and thus enhancing SUMO2/3ylation in rats.
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Objective To evaluate the relationship between the triggering receptor expressed on myeloid cells (TREM) and postoperative cognitive dysfunction (POCD) in elderly patients.Methods Eighty American Society of Anesthesiologists physical status Ⅰ-Ⅲ patients,aged 65-85 yr,weighing 50-80 kg,scheduled for elective total knee replacement under spinal-epidural anesthesia were enrolled in this study.Cerebrospinal fluid (CSF) was extracted after a catheter was successfully inserted into subarachnoid space.Blood samples from the cubital vein was collected before anesthesia induction (T0) and at 24 and 72 h after surgery (T1,2).The concentrations of TREM1 and TREM2 in CSF and plasma and tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in plasma were measured by enzyme-linked immunosorbent assay.The expression of TREM1,TREM2,IL-6 and TNF-α protein and mRNA in mononuclear ceils in peripheral blood was detected using real-time polymerase chain reaction.Neuropsychological test was performed in the the same time period at 1 day before surgery and 7 days after surgery,and the Z score was used to diagnose the development of POCD.The patients were divided into POCD group (P group) and non-POCD group (NP group) according to whether or not POCD happened after surgery.Results The incidence of POCD was 22％.Compared with group NP,the plasma TREM1 concentrations at T1,2 and plasma IL-6 and TNF-α concentrations at T2 were significantly increased,and the expression of TREM1 mRNA and TNF-α mRNA at T1,2 and IL-6 mRNA at T2 was up-regulated in group P (P＜0.05).There was no significant difference in plasma TREM2 concentrations at each time point between and within groups (P＞0.05).There was a higher consistency between plasma and CSF TREM1 concentrations (Cronbach's Alpha=0.784,P＜ 0.01) and a high consistency between plasma and CSF TREM2 (Cronbach's Alpha =0.935,P＜0.01).Conclusion Up-regulated expression of central and peripheral TREM1 is related to the development of POCD in elderly patients.
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Objective To investigate the effects of hydrogen-rich saline on hippocampal mammalian target of rapamycin (mTOR)/tau pathway after operation in aged rats.Methods One hundred fifty healthy male Sprague Dawley rats, aged 18 months, weighing 400-540 g, were randomly divided into 5 groups (n=30 each) using a random number table: control group (group C);sham operation group (group S);operation group (group O);hydrogen-rich saline treatment group (group HS);normal saline group (group NS).Splenectomy was performed in group O.Hydrogen-rich saline 1 ml/100 g was injected intraperitoneally at 5 min before splenectomy in group HS.Normal saline 1 ml/100g was injected intraperitoneally at 5 min before splenectomy in group NS.Morris water maze test was performed at 1, 3 and 7 days after surgery, and the escape latency and swimming distance were recorded.After the end of the test, the rats were sacrificed, and the brains were removed for examination of the pathological changes in the hippocampal CA3 region (under light microscope), and for determination of the expression of mTOR mRNA and tau mRNA (using reverse transcriptase polymerase chain reaction) and mTOR and pS396 tau (by Western blot).Results Compared with C and S groups, the escape latency and swimming distance were significantly prolonged, and the expression of mTOR protein and mRNA, tau mRNA and pS396 tau was up-regulated in O, HS and NS groups (P＜0.05).Compared with O and NS groups, the escape latency and swimming distance were significantly shortened, and the expression of mTOR protein and mRNA, tau mRNA and pS396 tau was down-regulated in group HS (P＜0.05).The pathological changes of the hippocampal tissues were mitigated in group HS when compared with group O.Conclusion The mechanism by which hydrogen-rich saline mitigates postoperative cognitive dysfunction may be associated with inhibited activation of hippocampal mTOR/tau pathway in aged rats.
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Objective:To stimulate the PBMCs of systemic lupus erythematosus patients with recombinant human high mobility group box1 (HMGB1) protein,observe the effect of HMGB1 on the expression of LC3Ⅱ/Ⅰ protein in active lupus nephritis and inactive lupus nephritis patients.Evaluate the effect of recombinant human high mobility group box 1 ( HMGB1 ) on the proliferation of PBMCs in patients with SLE.Methods:Western blot was used to detect the expression of LC 3II/I protein in PBMCs of active lupus ne-phritis and inactive lupus nephritis patients after stimulated by 1 μg/ml HMGB1 for 0,6,24 and 48 h.CCK-8 assay was used to detect the effects of PBMCs proliferation in patients with SLE after 1 μg/ml HMGB1 stimulation 72 hours.The statistical software SPSS17.0 was used to analyzed the results.Results: Western bolt showed an increasing expression of LC 3Ⅱ/Ⅰ protein in SLE patients after stimulated by HMGB1 ( P<0.05 ) , and this effect was time dependent.Compared with inactive lupus nephritis group , the increasing level of autophagy in active lupus nephritis group was more obviously.1 μg/ml HMGB1 could inhibit the proliferation of PBMCs in patients of SLE significantly ( P<0.001 ).Conclusion: HMGB1 may promote autophagy in SLE especially patients with active lupus nephritis and involved in the pathogenesis of SLE and lupus nephritis.Monoclonal antibodies targeting to HMGB 1 or modulators of mam-malian autophagy may provide new way for the treatment of SLE especially LN.
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Objective Role of melanocortin receptor 4 (MCAR) in excitatory amino acid release from rat astrocytes in spinal cord. Methods Astrocytes were isolated from the spinal cord of newborn pathogen-free Wistar rats ( 1-3 days after birth) and cultured in serum-free Neurobasal/B27 liquid culture medium. After 4 passages the primary cultured astrocytes were randomly divided into 3 groups (6 wells each): group Ⅰ control (group C); group Ⅱ the astrocytes were exposed to TNF-α 10 μg/L (group T) and group Ⅲ the astrocytes were exposed to TNF-α 10 μg/L and HS014 (selective MC4R antagonist) 1 μmol/L (group TH). The astrocytes were incubated at 37 ℃ for 3 h. The supernatant was collected for determination of glutamic acid (Glu) and aspartic acid (Asp)concentrations by HPLC-MS/MS. Results TNF-α significantly increased Glu and Asp release from astrocytes in group T as compared with group C. The Glu and Asp concentrations were significantly lower in group TH than in group T. Conclusion MG4R is involved in the excitatory amino acid release from astrocytes in the spinal cord.
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Background and purpose:In recent studies, the Notch-1 gene has been found to play an important role in the development of human glioblastoma.Short interfering RNA(siRNA) was used to silence the Notch-1 gene and block its expression.The objective was to determine whether siRNA targeting Notch-1 would inhibit the formation and growth of tumors in nude mice that modeled human glioblastoma.Methods:The human glioblastoma cell line TJ905 were first cultured and transfected with Notch-1 siRNA or nonsense siRNA by OligofectamineTM.The TJ905 cells were divided into 3 groups:the Notch-1 siRNA transfected group, nonsense siRNA transfected group and the control group.Each group's cells were subcutaneously injected into 5 nude mice.The nude mice(males, 3-4 weeks old) were given subcutaneous injections of either 0.2 mL of transfected siRNA or with normal TJ905 cells suspended at a 1?107 cells/mL concentration in a DMEM medium without serum.One week later, when the tumors were palpable, they were directly injected with the Notch-1 siRNA complex, nonsense siRNA complex, or PBS every 4 days for 20 days.Tumor sizes were measured every 3 days and calculated by the formula:volume(mm3) =1/2 length?(width)2.After a 20-day follow-up period, the mice were exterminated.Immunohistochemistry was used to determine the expression of Notch-1 gene.Results:The final tumor volume was less in nude mice injected with Notch-1 siRNA(1 203?206) mm3 compared mice injected with the nonsense siRNA injection(2 241?401) mm3, P