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Objective To investigate the regulating function of human gingival mesenchymal stem cell (GMSC) on the proliferation and differentiation of B cells and its underlying molecular mechanism. Methods GMSC were isolated and B cells were isolated from peripheral blood. GMSC or fibroblasts were co-cultured with B cells in vitro and assigned into the GMSC group and fibroblast group. The proliferation of B cells was detected in two groups. The expression of IgG1 and IgM in the cell supernatants was measured between two groups. The secretion of interleukin (IL)-6, Perforin, interferon (IFN)-γ and tumor necrosis factor (TNF)-α was compared between two groups. The expression levels ofIL-10 and transforming growth factor (TGF)-β in B cells were detected between two groups. The expression of PC-1 in B cells was measured in two groups. The signaling pathway involved with the regulating effect of GMSC on B cell function was investigated. The regulating effect of GMSC on the role of B cells in activating T cell function was assessed. Results Compared with the fibroblast group, the proliferation of B cells was significantly weakened in the GMSC group (P < 0.05). Co-culture of GMSC and B cells significantly inhibited the secretion of IgG1 and IgM from B cells and the secretion ofIL-6, Perforin, IFN-γ and TNF-α (all P < 0.05). Compared with the fibroblast group, the secretion of IL-10 and TGF-βwas significantly higher in the GMSC group (both P < 0.05). The expression level of PC-1 in the GMSC group was significantly down-regulated (P < 0.05). After adding ALK5, an inhibitor of TGF-β receptor, the inhibitory effect of GMSC upon B cells was significantly weakened (P < 0.05). Compared with the fibroblast group, the ability of B cells to activate and proliferate T cells was significantly attenuated in the GMSC group (P < 0.05). Conclusions GMSC can inhibit B cells and their mediated immune responses. The activation of B cells and other related functions can be suppressed through the TGF-β signaling pathway.
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Objective To investigate the effects of Epigallocatechin gallate (EGCG) on cardiac protection in diabetic rats and the expression of TGF-1/Smad3 signaling pathway. Methods The influence of clean level 40 SD rats, weight 200-220 g, divided into four random groups:control (Sham) group, diabetic cardiomyopathy model (DC) group, EGCG group, and metformin positive control group (Met).Post 8 weeks of high-fat-diet administration, the rats were injected intraperitoneally with STZ to establish the diabetes cardiomyopathy model. Upon successful model establishment, the EGCG group was intraperitoneally injected with EGCG and the cardiac function of the rats was measured after 28 days of drug administration. Then, the pathological results of the myocardial tissue were analyzed. Triglyceride (TG), total cholesterol (TC), glycosylated hemoglobin (HbA1 c), and blood glucose (FBG) concentrations were also measured. Further, the concentrations of superoxide dismutases (SOD), malondialdehyde (MDA), catalase (CAT), and glutathione peroxidase (GPX) in serum were measured by ELISA. The expression of TGF-β1 and Smad3 in kidney tissues of the rats was measured by Western blotting analysis. Results EGCG could reduce the glucose, lipid, and MDA levels in the blood of the diabetic rats, enhance cardiac systolic and diastolic functions, inhibit TGF-β1 and Smad3 protein expression, enhance the activity of SOD, CAT and GPX, and reduce myocardial tissue fibrosis. Conclusion EGCG can protect diabetic rat hearts by improving metabolic disorder, and its mechanism may be related to the oxidative-stress mediated by the TGF-β1/Smad3 signaling pathway.
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Objective To evaluate the protective effect of epigallocatechin gallate (EGCG) on lipopolysaccharide (LPS) -induced acute kidney injury (AKI) in rats and its underlying mechanisms. Methods Sprague-Dawley rats were randomly divided into the Sham group, AKI group, EGCG group and TLR4 group (n = 10 each). To establish the rat model of endotoxemia, serum creatinine (Cr) and urea nitrogen (BUN) levels were detected by biochemical assays; serum interlukin (IL) -6, IL-1β, IL-10, and TNF-α levels were detected by ELISA; kidney histopathology was examined by hematoxylin and eosin (HE) staining method; and expression of TLR4, Myd88 and nuclear factor-kappa B (NF-κB) in rat kidneys at both protein and mRNA levels was detected by Western blotting and qRT-PCR, respectively.Results Kidney injury increased significantly in AKI group compared to the sham group. Serum Cr, BUN, IL-6, IL-1β, and TNF-α levels significantly increased whereas IL-10 levels significantly decreased in AKI group compared to the sham group. Expression levels of TLR4, Myd88, and NF-κB also significantly increased at both protein and mRNA levels in AKI group compared to the sham group. Treatment with EGCG prior to induction of LPS-mediated AKI conferred protection against AKI by significantly reducing the expression of inflammatory markers such as, TLR4, Myd88, and NF-κB. Given TLR4 inhibitor based on this, the protective effect of EGCG on AKI was via inhibition of the TLR4/Myd88/NF-κB pathway. Conclusion EGCG exhibited a protective effect against LPS-induced AKI by inhibiting the activation of TLR4/Myd88/NF-κB pathway.
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Objective To investigate the mechanism via which artesunate regulates the invasion and metastasis of colon cancer cells and the expression of members of the TGF-β1/Smad4 signaling pathway. Methods The cell counting kit 8 (CCK8), nude mouse xenograft model, Transwell invasion assay, and flow cytometry were used to investigate the effect of artesunate on the invasion and metastasis of colon cancer cells. Western blotting and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) were used to detect the expression of TGF-β1 and Smad4 proteins and mRNA, respectively. Results Artesunate inhibited the growth of transplanted tumor, cell proliferation, and invasion and promoted apoptosis. It inhibited TGF-β1 expression and promoted Smad4 expression. TGF-β1 inhibitors reversed the inhibitory effect of artesunate. Conclusion Artesunate can inhibit the growth of xenograft tumor in nude mice and its mode of action may be related to the TGF-β1/Smad4 signaling pathway.
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Objective To investigate the effect of hydrogen-rich saline on apoptosis in hippocampal neurons induced by cerebral ischemia-reperfusion in rats and PI3K/Akt/FoxO1 signaling pathway.Methods The rat model of focal cerebral ischemia-reperfusion was established by thread-occlusion of the middle cerebral artery in rats.SD rats were randomly divided into sham operation group (Sham group), ischemia-reperfusion group (I/R group) and hydrogen-rich saline treatment group (HRS group), 10 rats in each group.At 24 h after reperfusion, the serum levels of IL-6, TNF-a and IL-1β were detected by ELISA.Histological changes of the hippocampus were observed by pathology using HE staining.Apoptosis in brain tissues was observed by TUNEL staining.The expression changes of p-PI3K, Akt, caspase-3 and FoxO1 proteins were detected by Western blot assay.Results Compared with the sham group, pyramidal cells were arranged loosely in the I/R group and a large number of pyramidal cells were necrotized, and the amount of apoptotic hippocampal cells was increased.The levels of IL-1β, IL-6 and TNF-α were significantly increased (P< 0.05), as well as the expression of p-PI3K, Akt and caspase-3 in the brain tissue.However, the expression of FoxO1 protein was decreased.There were significant differences between the two groups (P< 0.05).Compared with the I/R group, the inflammatory factors were significantly decreased in the HRS group.The expressions of p-PI3K, Akt and caspase-3 were also significantly decreased, while the expression of FoxO1 protein was increased (P< 0.05).Conclusions Hydrogen-rich saline can reduce brain injury caused by ischemia-reperfusion, and its mechanism may be related to PI3K/Akt/FoxO1 signaling pathway.
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Objective To evaluate the effect of hemorrhagic shock factor on the pharmacokinetics of rocuronium in pigs.Methods Sixteen pathogen-free Bama miniature pigs of both sexes,aged 3-5 months,weighing 22-25 kg,were divided into 2 groups (n=8 each) using a random number table:control group (group C) and hemorrhagic shock group (group HS).In group C,rocuronium 3.78 mg/kg was injected via the auricular vein.In group HS,the animals were subjected to volume-controlled hemorrhage,about 40％ of blood volume was withdrawn from the left femoral artery over 15 min (30 ml/kg),and rocuronium 3.78 mg/kg was injected via the auricular vein after the model was successfully established.At 0,2,4,7,10,15,20,30,60,120,180,240,300,360 and 420 min after rocuronium injection,blood samples were collected from the internal jugular vein for determination of the plasma concentration of rocuronium by high-performance liquid chromatography-tandem mass spectrometry.The pharmacokinetic parameters of rocuronium were calculated.Results Compared with group C,the plasma concentration of rocuronium was significantly increased at 20 and 60-420 min after rocuronium injection,the elimination half-life and mean residence time were prolonged,and the plasma effect-site equilibration rate constant was decreased in group HS (P＜0.05).There was no significant difference in the maximal concentration and area under the concentration-time curve between the two groups (P＞ 0.05).Conclusion The elimination of rocuronium is slower in a pig model of hemorrhagic shock.
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Objective To evaluate the effect of sevoflurane on activation of nuclear factor kappa B (NF-κB) during brain injury induced by hemorrhagic shock (HS) in pigs.Methods Thirty-two adult male Bama miniature pigs,aged 6 months,weighing 22-25 kg,were divided into 4 groups (n=8 each) using a random number table:sham operation group (group Sham),HS group,sevoflurane preconditioning group (group Sev-Pre) and sevoflurane postconditioning group (group Sev-Post).The animals were anesthetized,and tracheostomized and mechanically ventilated.In group Sham,the bilateral femoral arteries and internal jugular veins were only cannulated.HS was induced by removing 40％ of blood volume within 15 min (30 ml/kg) via the right femoral artery and maintaining at this level for 1 h before resuscitation in HS,Sev-Pre and Sev-Post groups.In group Sev-Pre,2％ sevoflurane was inhaled for 30 min,and then HS was induced.In group Sev-Post,2％ sevoflurane was inhaled for 30 min starting from the time point immediately after HS was induced.Immediately before establishment of the model and at 30,60,90,120,180 and 240 min of HS (T1-6),blood samples from the jugular vein were collected for determination of serum interleukin-1beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) concentrations by enzymelinked immunosorbent assay.At 4 h of HS,the rats were sacrificed,and brains were removed for microscopic examination of hippocampal CA1 region (using haematoxylin and eosin staining) and for determination of the expression of NF-κB in nucleoprotein (by Western blot).Results Compared with group Sham,the concentrations of serum IL-1β and TNF-α were significantly increased at T2-6,and the expression of NF-κB in nucleoprotein in hippocampal CA1 region was up-regulated in HS,Sev-Pre and Sev-Post groups (P＜0.05).Compared with group HS,the concentrations of serum IL-1β and TNF-α were significantly decreased at T3-6,and the expression of NF-κB in nucleoprotein in hippocampal CA1 region was down-regulated in Sev-Pre and Sev-Post groups (P＜0.05).There were no significant differences between group SevPre and group Sev-Post in concentrations of serum IL-1β and TNF-α and expression of NF-κB in nucleoprotein in hippocampal CA1 region (P＞0.05).The pathologic changes were significantly attenuated in SevPre and Sev-Post groups as compared with group HS.Conclusion The mechanism by which sevoflurane attenuates brain injury induced by HS may be related to inhibition of NF-κB activation and reduction of inflammatory responses in pigs.
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Objective To evaluate the effect of sevoflurane preconditioning on brain injury induced by cardiopulmonary bypass (CPB) in rats.Methods Forty adult male Sprague-Dawley rats,aged 6-8 months,weighing 350-450 g,were randomly divided into 5 groups (n=8 each) using a random number table:sham operation group (S group),CPB group,and preconditioning with different concentrations of sevoflurane groups (SP1,SP2 and SP3 groups).In SP1,SP2 and SP3 groups,sevoflurane with the final concentrations of 1.2％,2.4％ and 3.6％,respectively,was inhaled for 1 h,and then CPB was started.After sevoflurane preconditioning and before CPB (T0),at 30 min of CPB (T1),at the end of CPB (T2),and at 1,2 and 3 h after termination of CPB (T3-5),venous blood samples were collected from the right internal jugular vein for determination of serum S100-β protein concentrations by enzyme-linked immunosorbent assay.Rats were sacrificcd at T5,and hippocampi were isolated for determination of neuronal apoptosis (by TUNEL) and NF-κB p65 expression (by immunohistochemistry).Results Compared with group S,the concentration of serum S100-β protein was significantly increased at T1-5,the number of apoptotic neurons was significantly increased,and the expression of NF-κB p65 was significantly up-regulated in CPB,SP1,SP2 and SP3 groups (P＜0.05).Compared with group CPB,the serum S100-β protein concentration was significantly decreased at T1-5,the number of apoptotic neurons was significantly decreased,and the expression of NF-κB p65 was significantly down-regulated in SP1,SP2 and SP3 groups (P＜ 0.05).Compared with group SP1,the serum S100-β protein concentration was significantly decreased at T1-5,the number of apoptotic neurons was significantly decreased,and the expression of NF-κB p65 was significantly down-regulated in SP2 and SP3 groups (P＜0.05).Compared with group SP2,the serum S100-β protein concentration was significantly decreased at T1-5,the number of apoptotic neurons was significantly decreased,and the expression of NF-κB p65 was significantly downregulated in group SP3 (P＜0.05).Conclusion Sevoflurane preconditioning can attenuate CPB-induced brain injury probably by inhibiting activation of NF-κB in hippocampal neurons of rats.
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Objective To evaluate the effect of sevoflurane on brain injury in pigs with hemorrhagic shock (HS).Methods Twenty-four adult male Bama miniature pigs, aged 6 months, weighing 22-25 kg, were equally and randomly divided into 3 groups using a random number table: sham operation group (group Sham) , group HS, and sevoflurane group (S group).In group Sham, the bilateral femoral arteries and internal jugular vein were only punctured.The animals were anesthetized with iv propofol 3.0 mg/kg, tracheostomized and mechanically ventilated.The right femoral artery was cannulated for blood-letting.HS was induced by blood-letting (40％ blood volume within 15 min), and it was then maintained for 1 h after the end of blood-letting to induce brain injury.In group S, 2％ sevoflurane was inhaled for 30 min after successful establishment of the model.Immediately before establishment of the model (T0) , and at 30, 60,90, 120, 180 and 240 min after HS (T1-6) , blood samples were collected from the internal jugular vein for determination of interleukin-1beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) concentrations in serum (by enzyme-linked immunosorbent assay), and neuron-specific enolase (NSE) and S-100β protein concentrations in serum (using double antibody sandwich method).Results Compared with group Sham, the serum IL-1β, TNF-α, NSE and S-100β protein concentrations were significantly increased at T2-6 in HS and S groups (P＜0.05).Compared with group HS, the serum IL-1β, TNF-α, NSE and S-100β protein concentrations were significantly decreased at T3-6 in group S (P＜ 0.05).Conclusion Sevoflurane can mitigate brain injury in pigs with HS, and the mechanism is associated with inhibition of inflammatory responses.
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Objective To evaluate the changes in the expression of aquaporin-8 (AQP8) in intestinal mucosa in pigs with hemorrhagic shock.Methods Sixteen Bama miniature pigs,weighing 22-25 kg,were equally and randomly divided into sham operation group (group S) and hemorrhagic shock group (group HS).The animals were fasted for 8 h before operation.The animals were anesthetized with propofol 3 mg/kg injected via the auricular vein,and tracheostomized and mechanically ventilated.In group S,the femoral artery and internal jugular vein were only cannulated.In group HS,the femoral artery and internal jugular vein were cannulated for blood pressure and mean arterial pressure monitoring and blood sampling.Hemorrhagic shock was then induced by removing 40 percent of blood volume over 15 min.Before anesthesia (T0),and at 30 min and 1.0,1.5,2.0,3.0 and 4.0 h after the end of blood-letting (T1.6),blood samples were collected for determination of serum D-lactate and intestinal fatty acid binding protein (I-FABP) concentrations.After blood sampling at T6,the pigs were sacrificed,and intestinal specimens were obtained for microscopic examination and for determination of AQP8 cotent in intestinal mucosa (by ELISA).The water content of intestines was calculated by wet/dry weight ratio.Results Compared with group S,the serum D-lactate concentrations at T2-6,I-FABP concentrations at T1-6,and water content of intestines were significantly increased,and the cotent of AQP8 was up-regulated at T6 in group HS.No changes were found in the intestinal mucosa in group S.In group HS,severe damage to the intestinal mucosa was found,and bleeding,inflammatory cell infiltration,and epithelial cell necrosis were observed.Conclusion The mechanism of hemorrhagic shock-caused damage to intestines is related to up-regulated expression of AQP8 in intestinal mucosa in pigs.
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Objective To evaluate the effect of sevoflurane on myocardial injury induced by hemorrhagic shock and resuscitation (HS/R) in pigs.Methods Twenty-four Bama miniature pigs (12 males, 12 females) , weighing 20-25 kg, aged 3-5 months, were randomly divided into 3 groups (n=8 each) using a random number table: sham operation group (group S) , HS/R group and sevoflurane group (group Sev).The left and right femoral arteries and right femoral vein were cannulated for blood pressure monitoring, blood-letting, blood sampling and fluid infusion.HS/R was induced by blood-letting maintaining for 1 h, followed by resuscitation with autologous blood reinfusion and infusion of lactated Ringer's solution 2 times the volume of the blood withdrawn.The pigs in group Sev were exposed to 2％ sevoflurane for 30 min before resuscitation.After cannulation, at 30 min after hemorrhagic shock, before resuscitation, and at 30 min, and 1.5, 2.5 and 3.5 h after resuscitation, blood samples were collected from the femoral artery for determination of creatine kinase-MB (CK-MB) activity and cardiac troponin Ⅰ (cTnI) concentration in serum using an automatic biochemical analyzer.Myocardial specimens were obtained at 3.5 h after resuscitation for detection of tumor necrosis factor-alpha (TNF-ot) and interleukin-6 (IL-6) contents (by ELISA) , and phosphor-signal transducer and activator of transcription 1 (p-STAT1) expression (by Western blot), and for examination of the pathological changes (with light microscope).Results Compared with S group , the CK-MB activity and cTnI concentration in serum and contents of TNF-α and IL-6 were significantly increased, and the expression of p-STAT1 was up-regulated in HS/R and Sev groups (P＜0.05).Compared with HS/R group, the CK-MB activity and cTnI concentration in serum and contents of TNF-α and IL-6 were significantly decreased, and the expression of p-STAT1 was downregulated (P＜0.05) , and the pathological changes of myocardia were alleviated in Sev group.Conclusion Sevoflurane can alleviate HS/R-induced damage to myocardia of pigs, and inhibited STAT1 activity and attenuated inflammatory responses in the myocardium are involved in the mechanism.
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Objective To investigate the effect of dexmedetomidine on acute kidney injury in endotoxemic rats.Methods Thirty adult male Sprague-Dawley rats,aged 4-6 months,weighing 180-220 g,were randomly divided into 3 groups (n =10 each) using a random number table:control group (group C),lipopolysaccharide group (group L),and dexmedetomidine (group D).Lipopolysaccharide (LPS) 5 mg/kg was injected slowly into the femoral vein to establish the model of endotoxemic in rats anesthetized with chloral hydrate.In group D,after LPS injection,a loading dose of dexmedetomidine 7 μg/kg was injected intravenously,and 15 min later dexmedetomidine was infused for 6 h at 5 μg · kg-1 · h-1,while the equal volume of normal saline was given in L and C groups.At 6 h after the end of LPS administration,blood samples were collected from the femoral vein for determination of serum creatinine (Cr),blood urea nitrogen (BUN),tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) concentrations.At 24 h after the end of LPS administration,the animals were sacrificed and kidneys were removed for microscopic examination and for determination of the expression of tight junction proteins ZO-1 and occludin in renal tissues by Western blot.Results Compared with group C,the serum Cr,BUN,TNF-α and IL-6 concentrations were significantly increased,and the expression of ZO-1 and occluding was down-regulated in L and D groups.Compared with group L,the serum Cr,BUN,TNF-α and IL-6 concentrations were significantly decreased,the expression of ZO-1 and occluding was up-regulated,and the pathological changes of kidneys were mitigated in D group.Conclusion Dexmedetomidine can alleviate acute kidney injury in endotoxemic rats.
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Objective To evaluate the effect of dexmedetomidine on lung injury induced by renal ischemia/reperfusion (l/R) in rats.Methods Healthy male Sprague-Dawley rats,aged 4-5 months,weighing 250-300 g,were randomized into 4 groups (n =10 each) using a random number table:sham operation group (group S); group I/R; dexmedetomidine pretreatment group (group D1) and dexmedetomidine postconditioning group (group D2).Renal I/R was induced by right nephrectomy and occlusion of the left kidney for 45 min followed by reperfusion in animals anesthetized with intraperitoneal chloral hydrate.In group D1,dexmedetomidine was infused intravenously starting from 30 min before ischemia until beginning of ischemia.In group D2,starting from onset of reperfusion until 30 min of reperfusion,dexmedetomidine was infused intravenously for 10 min at a rate of 1 μg· kg-1 · h-1,and then infused for 20 min at 0.5 μg· kg-1 · h 1.Blood samples were collected at 6 h of reperfusion to determine serum creatinine,blood urea nitrogen,interleukin-1β (IL-1β),IL-6 and tumor necrosis factor-α (TNF-α) concentrations,and IL-1β,IL-6 and TNF-α concentrations in broncho-alveolar lavage fluid (BALF).Lungs were removed for microscopic examination and for determination of wet/dry lung weight ratio.Results Compared with group S,wet/dry lung weight ratio,serum creatinine and blood urea nitrogen concentrations,and IL-1β,TNF-α and IL-6 concentrations in serum and BALF were significantly increased in the other three groups (P ＜ 0.05).The parameters mentioned above were significantly lower in D1 and D2 groups than in I/R group (P ＜ 0.05).Microscopic examination showed that the pathological changes were significantly attenuated in D1 and D2 groups as compared with I/R group.Conclusion Both dexmedetomidine pretreatment and postconditioning can attenuate lung injury induced by renal I/R and inhibition of inflammatory responses is involved in the mechanism.