The study on inflammatory mechanism of cognitive dysfunction induced by 1-bromopropane in rats / 中华行为医学与脑科学杂志

Objective:To observe the role of neuroinflammation in cognitive dysfunction induced by 1-bromopropane (1-BP) in rats.Methods:Male Wistar rats were randomly divided into control group, 1-BP group, pyrrolidine dithiocarbamate(PDTC)+ 1-BP group and PDTC group, with 15 rats in each group. Rats in 1-BP group and PDTC+ 1-BP group were given 800 mg / kg 1-BP by gavage, and rats in control group and PDTC group were given equal volume corn oil once a day for 12 days; rats in PDTC group and PDTC+ 1-BP group were intraperitoneally injected with 100 mg / kg PDTC 30 minutes after gavage, while rats in control group and 1-BP group were injected with equal volume of normal saline once a day for 12 days.From the 7th to 12th day of the experiment, ten rats in each group were randomly selected and subjected to Morris water maze test for detect the cognitive function. In the positioning navigation test, the learning ability of rats was evaluated by the escape latency and total swimming distance respectively. In the space exploration experiment, the memory ability of experimental animals was evaluated by the number of times crossing the target platform. After the experiment, ten rats were sacrificed, the cerebral prefrontal cortex was harvested. The cytosolic and nuclear NF-κB expression and phosphorylation were detected by Western blot, the mRNA levels of TNF-α and IL-1β were detected by qRT-PCR. After cardiac perfusion fixation, the brains of 5 rats were taken to make frozen sections for immunohistochemical staining and Nissl staining. SPSS 20.0 software was used for statistical analysis, repetitive measurement deviation analysis was used for the analysis of the swimming distance and the escape latency in positioning navigation test, One-way ANOVA was used for the analysis of the number of times crossed the target platform in spatial probe test and other data. Tukey's test was used for Post hoc comparison.Results:The results of Morris water maze showed that there was significant interaction between group and training time in the total swimming distance of rats in the four groups ( F=3.762, P<0.05). Simple effect analysis showed that the total swimming distance of 1-BP group in 1-4 days were longer than those of control group (all P<0.05), while the total swimming distance of PDTC+ 1-BP group in 1-4 days were shorter than those of 1-BP group (all P<0.05). There was significant interaction between group and training time in the escape latency among the four groups ( F=6.541, P<0.01). The escape latencies of 1-BP group in 1-4 days were longer than those of control group (all P<0.05), while the escape latencies of PDTC+ 1-BP group in 1-4 days were shorter than those of 1-BP group (all P<0.05). The results of space exploration experiment showed that there was significant difference in the number of crossing the platform among the four groups ( F=75.333, P<0.01). The number of crossing the platform (1.08±0.29) in 1-BP group was lower than that in the control (3.35±0.05) ( P<0.01). The number of crossing the platform (1.95±0.26) in PDTC+ 1-BP group was higher than that in 1-BP group ( P<0.01). It had significant difference both in the cytoplasm and in the nucleus of the NF-κB protein level in prefrontal cortex among rats of the four groups ( F=20.865, 23.877, both P<0.01). The levels of NF-κB in cytoplasm and nucleus of rats in 1-BP group were both higher than those in control group (cytoplasm: (177.3±32.1)%, (100.0±8.4)%, P<0.01; nucleus: ( 173.2±27.1)%, (100.0±8.4)%, P<0.01). While the levels of NF-κB in cytoplasm and nucleus of 1-BP+ PDTC group were both lower than those of 1-BP group (cytoplasm: (148.7±22.0)%, (177.3±32.1)%, P<0.01; nucleus: (149.7±18.8)%, (173.2±27.1)%, P<0.01). The results of qRT-PCR showed that there were significant differences in the mRNA levels of TNF-α and IL-1β in the prefrontal cortex among the four groups ( F=17.464, 17.382, both P<0.01). The levels of TNF-α and IL-1β mRNA in 1-BP group were higher than those in control group (both P<0.05), and the levels of TNF-α and IL-1β mRNA in PDTC+ 1-BP group were both lower than those in 1-BP group (both P<0.05). The results of immunohistochemistry showed that compared with the control group, the number of microglia and astrocytes in the 1-BP group increased (microglia: (158.30±9.68), (110.20±16.30), P<0.05; astrocytes: (122.76±4.35), (80.24±6.96), P<0.05), and the morphology was also activated, with light staining and reduced number of Nissl bodies in neurons.The number of microglia and astrocytes in PDTC + 1-BP group was lower than that in 1-BP group (microglia: (131.70±14.67), (158.30±9.68), P<0.05; astrocytes: (101.54±4.55), (122.76±4.35), P<0.05), and the Nissl body staining of neurons was significantly deepened. Conclusion:NF-κB signaling pathway might be the key mechanism of 1-BP neurotoxicity. PDTC intervention could significantly improve the neuroinflammatory response and behavioral disorders of experimental animals intoxicated with 1-BP.

The effect of NF-κB on NLRP3 of lung tissue in rats with acute lung injury induced by phosgene / 中华急诊医学杂志

Objective To observe the effect of signal transduction pathway of nuclear factor-kappa B (NF-κB) on Nod-like receptor family, pyrin domain-containing 3 (NLRP3) inflammasome and pyroptosis in rats with acute lung injury induced (ALI) by phosgene. Methods The rats were randomly(random number) divided into 3 groups: air exposure control group, phosgene exposure group and PDTC group with phosgene exposure after 100 mg/kg pyrrolidine dithiocarbamate (PDTC) administration. The specimens of serum, bronchoalveolar lavage fluid (BALF) and lung tissue were collected 6 h after exposure. Morphological changes were observed by HE staining. The expression of NLRP3 in the lung of three groups was detected by immunohistochemistry. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expressions of NF-κB p65, NLRP3, ASC and caspase-1 mRNA in the lung tissue. NF-κB p65,NLRP3, ASC and caspase-1 protein levels in the lung tissue were quantified by Western blot. The concentrations of IL-1β, IL-18 and IL-33 in the serum and BALF were measured by enzyme-linked immunosorbent assay (ELISA). Pyroptosis was observed by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL). Results The model of phosgene-induced ALI was successfully established in rats. Morphological changes with inflammatory cell infiltration were observed in the lung tissues of phosgene group, in which NLRP3 positive cells also could be observed by immunohistochemical staining. The mRNA expressions and protein levels of NF-κB p65, NLRP3 and caspase-1 in lung tissues were significantly increased (P＜0.05) in phosgene group, compared with air control group. The mRNA expressions and protein levels of NF-κB p65,NLRP3 and caspase-1 in lung tissues were significantly decreased (P＜0.05) in PDTC group, compared with phosgene group. The IL-1β,IL-18 and IL-33 protein levels in serum and BALF were significantly increased (P＜0.05) in phosgene group, compared with air control group. The IL-1β,IL-18 and IL-33 protein levels in serum and BALF were significantly decreased (P＜0.05) in PDTC group, compared with phosgene group. TUNEL results showed that pyroptosis in the lung tissue obviously increased in phosgene group, while decreased in PDTC group. Conclusions NLRP3 inflammasome and lung cell pyroptosis were induced through NF-kB signal transduction pathway in rats with acute lung injury caused by phosgene inhalation. Blockade of NF-κB can alleviate acute lung injury by down-regulating the expression of NLRP3 inflammasome to inhibit pyroptosis.

Abnormality of coagulation and fibrinolysis functions of endothelial cells induced by activated complement alternative pathway and intervention / 中国药理学通报

Aim To investigate the change of coagulation and fibrinolysis functions of human microvascular endothelial cells (HMEC) induced by activated complement alternative pathway and effect of pyrrolidine-dithiocarbamate (PDTC) and resveratrol (Res) on intervention.Methods Normal human serum was activated by cobra venom factor (CVF).After exposure of HMEC to activated complement for various time points, supernatant was removed and assayed for activities of hydrolysing chromogenic substrate and affecting activated partial thromboplastin time (APTT) and prothrombin time (PT).The cells exposed to activated complement were collected and washed, and then the cell suspension was assayed for activity of affecting coagulation function of normal plasma.Lastly, the coagulation and fibrinolysis functions of HMEC pretreated with PDTC and Res were also investigated after HMEC was exposed to activated complement alternative pathway.Results The hydrolysis activity of chromogenic substrate of supernatant was up-regulated significantly after HMEC exposed to activated complement alternative pathway.The supernatant induced APTT decreased significantly, and also shortened PT.The cell suspension of various time points induced APTT decreased significantly, and also shortened PT by suspension of 6 h time point.PDTC and Res failed to inhibit the up-regulation of the chromogenic hydrolysis activity, but Res showed significant intervention on decrease of APTT, and PDTC had better effect on inhibiting the decrease of PT than that of Res.Conclusion Activated complement alternative pathway can induce abnormality of coagulation and fibrinolysis functions of HMEC, and PDTC and Res can affect this change.

Effects of PDTC and curcumin on expression of tissue factor induced by anti-β2GPI in mice / 中国免疫学杂志

Objective:To investigate whether PDTC or curcumin had effect on anti-β2GPI-induced tissue factor (TF) expression in mice.Methods:BALB/c mice were pretreated with PDTC (100 mg/kg,once a day) by intraperitoneal injection (i.p.) or/and curcumin (50 mg/kg,once a day) by oral gavage for 3 consecutive days at 2 h before 500 μg of anti-β2GPI injections in subsequent experiments.Mouse peritoneal macrophages and aorta were collected,homogenized by sonication.The total RNA and protein were collected from each animal,TF expression was detected by Real-time quatitative PCR and TF activity kit.The phosphorylation of NF-κB p65 and c-Jun/AP-1 was determined by Western blot.Results:Anti-β2GPI cloud significantly upregulate TF expression and phosphorylation of NF-κB p65 and c-Jun/AP-1 in mouse peritoneal macrophages and aorta,compared with NR-IgG treated mice (P＜ 0.05).PDTC or/and curcumin could markedly attenuate anti-β2 GPI-induced TF expression,also inhibit activation of NF-κB p65 and cJun/AP-1 in the aorta and peritoneal macrophages respectively (P＜0.05),but combination of two inhibitors had no synergistic effect.Conclusion:Both PDTC and curcumin could affect the expression of TF induced by anti-β2GPI in mice,indicatiug that PDTC and curcumin has the potential to prevent thrombosis in APS.

Effect of NF-κB inhibitor pyrrolidine dithiocarbamate on proliferation and apoptosis of human multiple myeloma U266 cells / 中国病理生理杂志

AIM: To explore the effect of pyrrolidine dithiocarbamate (PDTC), an NF-κB inhibitor, on the proliferation and apoptosis of human multiple myeloma U266 cells and its mechanisms.METHODS: The U266 cells were treated with PDTC at different concentrations (0, 25, 50, 100 and 200 μmol/L) in vitro.The growth inhibitory rate of the U266 cells was detected by CCK-8 assay and cell counting.The cell cycle of the U266 cells was determined by flow cyto-metry, and the apoptosis was examined by flow cytometry with Annexin V-FITC/PI staining.The effect of PDTC on the expression of DNA methyltransferase 1 (DNMT1) at mRNA and protein levels was measured by RT-qPCR and Western blot, respectively.The effects of PDTC on the protein levels of NF-κB (P65), DNMT1, Bcl-2, cyclin D1, cleaved caspase-3 and cleaved caspase-8 were determined by Western blot.RESULTS: The protein level of NF-κB (P65) was decreased after treatment with PDTC for 48 h or 72 h.PDTC inhibited the proliferation of U266 cells in both dose-and time-dependent manners.After treatment with PDTC for 48 h, the percentage of U266 cells in G2 phase increased compared with control group (P<0.05).PDTC induced the apoptosis of U266 cells in a dose-dependent manner.The expression of DNMT1 at mRNA and protein levels decreased (P<0.05).The results of Western blot showed that the expression of Bcl-2 in PDTC groups decreased, while the protein levels of cyclin D1, cleaved caspase-3 and cleaved caspase-8 were higher than those in control group (P<0.05).CONCLUSION: The NF-κB inhibitor PDTC inhibits the proliferation of U266 cells by inducing cell apoptosis.It may be related to the down-regulated expression of DNMT1, cell cycle arrest and activation of the apoptotic pathways.

Effect of NF- κ B inhibitor PDTC on VEGF and endostatin expression of mice with Lewis lung cancer

OBJECTIVE@#To investigate the effects of NF- κ B inhibitor pyrrolidine dithiocarbamate hydrochloride (PDTC) on vascular endothelial growth factor (VEGF) and endostatin expression in mice with Lewis lung cance; and its mechanism.@*METHODS@#Mice survival rate and anti-tumor effects were observed in different concentrations of NF- κ B inhibitor PDTC after the Lewis lung cancer mice model was established. VEGF and endostatin expressions were detected by immunohistochemical assay.@*RESULTS@#Lewis lung cancer was be inhibited by 0.5 mg/kg, 1.5 mg/kg and 3.0 mg/kg of NF- κ B inhibitor PDTC (P<0.05). Microvessel density (MVD) in 0.5 mg/kg, 1.5 mg/kg and 3.0 mg/kg NF- κ B inhibitor PDTC groups were significantly lower than the control group (P<0.05). Immunohistochemical assay results showed that VEGF and endostatin expressions in the 0.5 mg/kg, 1.5 mg/kg and 3.0 mg/kg NF-κ B inhibitor PDTC groups were significantly lower than the control group (P<0.05). Western blot results also showed that NF- κ B inhibitor PDTC could inhibit VEGF and endostatin expressions in tumor tissues.@*CONCLUSIONS@#NF- κ B inhibitor PDTC can inhibit tumor formation and reduce tumor angiogenesis in mice with Lewis lung cancer; and its mechanism maybe associated to VEGF and endostatin down-regulation.

Effect of blunting NF kappa B activation on CYP2 E1 in immunological liver injury rats / 中国药理学通报

Aim To determine the function of nuclear factor-κB ( NF-κB ) in immunological liver injury of rat model and its effect on CYP2E1 expression, content and metabolic activity. Methods The immunological liver injury rat model was prepared by injection of Ba-cillus Calmette Guérin ( BCG,125 mg · kg-1 ) for 14 days. The hepatic tissue injury was revealed by hema-toxylin and eosin ( HE ) method and serum concentra-tion of alanine aminotransferase( ALT) , aspartate ami-notransferase ( AST ) respectively. CYP450 total con-tent in hepatic homogenate was determined by spectro-photography. The expression of CYP2E1 protein was detected by Western blot analysis. The enzyme kinetics of CYP2 E1 probe drug chlorzoxazone was evaluated by high-performance liquid chromatography ( HPLC ) as-say. Results The results showed that BCG-pretreat-ment ( 125 mg · kg-1 ) significantly increased the weight of liver and spleen, serum levels of ALT and AST(P<0. 01) , and decreased CYP2E1 expression, content and metabolic activity ( P <0. 05 ) . Adminis-tration of ammonium pyrrolidine dithiocarbamate (PDTC) (50, 100, 200 mg·kg-1) reversed the a-bove hepatic injury stimulated by BCG in vivo. Moreo-ver, PDTC dose-dependently inhibited the down regu-lation of CYP2 E1 ( P<0. 05 ) . Conclusion Passiva-tion of NF-κB can inhibit the down regulation of CYP2 E1 in liver tissue of immunological liver injury rats;NF-κB may be involved in CYP2 E1 down-regula-tion.

Expression of coagulation-and fibrinolysis-related molecules of endothelial cells induced by activated complement alternative pathway and intervention / 中国药理学通报

Aim To investigate the change of molecu-lar expression related to coagulation and fibrinolysis in human microvascular endothelial cells ( HMEC ) in-duced by activated complement alternative pathway and effect of pyrrolidine dithiocarbamate ( PDTC ) and res-veratrol on intervention. Methods Normal human se-rum was activated by cobra venom factor ( CVF) . After exposure of HMEC to activated complement for various times, supernatant was removed and assayed for ex-pressions of P-selectin, VWF, t-PA, PAI-1, TF, TM, and NO by using reagent kits. The expressions of the above molecules in HMEC pretreated with PDTC and resveratrol were also investigated. Results P-selectin and VWF were rapidly released by endothelial cells and the expression reached the peak at the time point of 15 min. The expressions of t-PA, PAI-1, and TF were continuously upregulated, whereas NO and TM were decreased. PDTC and resveratrol inhibited the upregulation of P-selectin, VWF, t-PA, PAI-1 and TF, and intervened the downregulation of NO. Res-veratrol further downregulated the expression of TM. Conclusion Activated complement alternative path-way can influence the expression of molecules related to coagulation and fibrinolysis in HMEC, and PDTC and resveratrol can affect this change.

Effect of antioxidants on pancreatic fibrosis of rats with chronic pancreatitis / 中华胰腺病杂志

Objective To investigate the effect of antioxidants including PDTC on pancreatic fibrosis of rats with chronic pancreatitis.Methods The rats were randomly divided into 5 groups including control group, CP group, PDTC treatment group, vitamin E treatment group and vitamin C treatment group.The CP model was in ducad by using intraperitoneal injection of DETC (750 mg/kg), twice a week.The control group received no treatment.After DETC injection, the treatment groups received an intraperitoneal injection of PDTC (100 mg/kg), vitamin E (15 mg/kg), vitamin C (15 mg/kg), respectively.Rats were sacrificed at 90 min, 24 h, 48 h, 72 h, 2 w, 3 w, 4 w, 6 w after first injection of DETC.Pancreatic tissue was taken for routine pathological examination.The activity of SOD, GSH-PX and MDA content were detected by spectrophotometric ratio method.α-SMA, desmin collagen Ⅰ, Ⅲ, TGF-β1, FN were detected by immunohistochemical assay.The expression of TGF-β1, FN mRNA was measured by RT-PCR.Results At 6w, the fibrosis and the parameters for damage of the pancreas in the three treatment groups were significantly better than that in CP group (P ＜0.01), the vacuolar degeneration index in vitamin E group and vitamin C group was also better than that in CP group (P ＜0.01).From the 2nd week, the activity of SOD, GSH PX in PDTC group, Vit C group and Vit E group was higher than that in CP group, while the MDA activity was lower than that in CP group, and the difference was statistically significant (P ＜ 0.01 or P ＜ 0.05).No significant difference was found among the three treatment groups.The mRNA levels of TGF-β1 and FN of the treatment groups were lower than those of CP group (P ＜0.05 or P ＜0.01), but higher than those of the control group (P ＜ 0.05).There was no significant difference among the three treatment groups (P ＞ 0.05).Conclusions PDTC and the other antioxidants can reduce oxygen free radicals by increasing the activity of SOD,suppressing the activation of PSCs, reducing the secretion of TGF-β1, Collagen Ⅰ , Ⅲ, FN and eventually inhibit the progress of pancreatic fibrosis.

Effect of NF- κ B inhibitor PDTC on VEGF and endostatin expression of mice with Lewis lung cancer

Objective: To investigate the effects of NF- κ B inhibitor pyrrolidine dithiocarbamate hydrochloride (PDTC) on vascular endothelial growth factor (VEGF) and endostatin expression in mice with Lewis lung cance; and its mechanism. Methods: Mice survival rate and anti-tumor effects were observed in different concentrations of NF- κ B inhibitor PDTC after the Lewis lung cancer mice model was established. VEGF and endostatin expressions were detected by immunohistochemical assay. Results: Lewis lung cancer was be inhibited by 0.5 mg/kg, 1.5 mg/kg and 3.0 mg/kg of NF- κ B inhibitor PDTC (. P<0.05). Microvessel density (MVD) in 0.5 mg/kg, 1.5 mg/kg and 3.0 mg/kg NF- κ B inhibitor PDTC groups were significantly lower than the control group (. P<0.05). Immunohistochemical assay results showed that VEGF and endostatin expressions in the 0.5 mg/kg, 1.5 mg/kg and 3.0 mg/kg NF-. κ B inhibitor PDTC groups were significantly lower than the control group (. P<0.05). Western blot results also showed that NF- κ B inhibitor PDTC could inhibit VEGF and endostatin expressions in tumor tissues. Conclusions: NF- κ B inhibitor PDTC can inhibit tumor formation and reduce tumor angiogenesis in mice with Lewis lung cancer; and its mechanism maybe associated to VEGF and endostatin down-regulation.

COX2-PGI2/TXA2 signal pathway involved in protective mechanism of PDTC pretreatment against global cerebral ischemia reperfusion rat hippocampus injury / 中国药理学通报

Aim To investigate the effects and mecha-nism of nuclear factor-κ B inhibitor, PDTC, on global cerebral ischemia reperfusion ( GCIR ) rat hippocam-pus. Methods Forty-eight adult male Sprague-Daw-ley rats were randomly divided into one control group receiving sham operation and three experimental groups all receiving global cerebral ischemia for 20 min. In PDTC 100 mg·kg-1 group ( P100 ) and PDTC 200 mg ·kg-1 group ( P200 ) , PDTC 100 mg · kg-1 or PDTC 200 mg·kg-1 was injected ip one hour before ischemi-a respectively. Spatial learning and memory function of rats were tested using Morris water maze. HE staining was employed to observe pathological changes of hipp-ocampal neurons. Expression of COX2 was measured by Western blot, and the content of PGI2 and TXA2 in rat hippocampus was detected by enzyme-linked immu-nosorbent assay. Results A significant increase of es-cape latency was observed in GCIR group compared to the sham operation group(P<0.05). PDTC 100 mg· kg-1 and PDTC 200 mg · kg-1 significantly reduced escape latency ( P <0.05 ) and histopathological injury in CA1 region of hippocampus. PDTC 100 mg · kg-1 and PDTC 200 mg · kg-1 also reduced COX2 expres-sion, PGI2 content, TXA2 content and PGI2/TXA2 . Conclusion Pretreatment with PDTC can protect hip-pocampus from GCIR injury through inhibition of COX2 expression and PGI2/TXA2 .

Effects of pyrrolidine dithiocarbamate on nuclear factor-κB/interleukin-6 signaling pathway in rats with chronic mixed reflux esophagitis / 中华消化杂志

Objective To observe the protective effects of nuclear factor (NF) κB inhibitor pyrrolidine dithiocarbamate (PDTC) on chronic mixed reflux esophagitis in rats and its influence on NF-κB/interleukin (IL)-6 signaling pathway.Method A total of 40 healthy male Sprague-Dawley (SD) rats were divided into healthy control group,sham operation group,model control group,omeprazole group and PDTC group with eight rats in each group.Except rats in healthy control group and sham operation group,mixed reflux esophagitis model were established in all the other groups.The rats of healthy control group,sham operation group and model control group were all intraperitoneally injected with 2 mL 0.9％ NaCl,rats of omeprazole group were intraperitoneally injected with omeprazole 20 mg/kg,and rats of PDTC group were intraperitoneally injected with PDTC 100 mg/kg every day.After six weeks,the rats were sacrificed,the morphological changes of esophageal tissues were observed and scored by visual inspection and under light microscope.The serum levels of NF-κB p65 and IL-6 in rats of each group were assessed by enzyme linked immunoassay (ELISA).t test was performed for mean comparison among groups.Results The scores of esophageal mucosa judged by visual inspection of healthy control group,sham operation group,model control group,omeprazole group and PDTC group were 0.000 20.000,0.000±0.000,2.250± 0.707,1.125 ± 0.835 and 1.429± 0.535,respectively.The pathological scores were 0.00020.000,0.000±0.000,2.625±0.518,1.500±0.535,1.429±0.535,respectively.Compared with those of model control group,the scores judged by visual inspection and the pathological scores of healthy control group,sham operation group,omeprazole group and PDTC group were lower,and the differences were statistically significant (t=7.603,7.603,2.909,2.506; t=9.674,9.674,4.277,4.399,all P＜0.05).The serum levels of NF-κB p65 protein of healthy control group,sham operation group,omeprazole group and PDTC group were (68.618±18.450) pg/mL,(77.824±22.228) pg/mL,(106.693±45.312) pg/mL and (103.781± 42.502)pg/mL,respectively; compared with that of model group ((184.882±49.165) pg/mL),which were significantly lower and the differences were statistically significant (t=6.262,5.612,3.308 and 3.427,all P＜0.05).The serum levels of IL-6 protein were (24.826±4.008) pg/mL,(23.599±4.351) pg/mL,(32.370± 11.657) pg/mL and (33.694±10.394) pg/mL,respectively,which significantly decreased when compared with that of model group ((51.378±9.697) pg/mL,t=7.157,7.393,3.546 and 3.392,all P＜0.05).There was no significant difference between PDTC group and omeprazole group in the score judged by visual inspection,pathological scores,the serum levels of NF-κB p65 and IL-6 protein (all P＞0.05).Conclusion NF-κB inhibitor PDTC could reduce the injury severity of esophageal mucosal in reflux esophagitis rat,which mechanism might be related with the down-regulation of NF-κB/1L-6 signaling pathway.

Effects of pyrrolidine dithiocarbamate on expression of NF-κB,MMP-2 and collagen remodeling after myocardial infarction in rats / 重庆医学

Objective Toinvestigatetheeffectsofpyrrolidinedithiocarbamate(PDTC)onexpressionofNF-κB,MMP-2andleft ventricular collagen remodeling following acute myocardial infarction in rats .Methods The myocardial infarction model in rat was induced by ligation of left anterior descending coronary artery .12 adult Sprague-Dawley rats survived 24 for h after acute myocardial infarction were randomly divided into the myocardial infarction (MI) group and the PDTC-treated(PD) group .Six rats were desig-nated as sham-operated group(SH group) .The PD group was intraperitoneally injected with PDTC (80 mg · kg -1 · d-1 ) for 28 d , the MI group and SH group were given normal saline as control .On 28 d ,the cardiac function of left ventricle was measured by ech-ocardiography .The infarct size was evaluated .The total collgen ,typeⅠcollgen ,typeⅢcollgen ,and Ⅰ /Ⅲ collgen ratio were quanti-fied histomorphometry .The mRNA and protein levels of NF-kappaBp65 and MMP-2 were determined by reverse transcription-poly-merase chain reaction(RT-PCR) and by Western blot ,respectively .Results Compared with the SH group ,the values of the total collgen ,typeⅠcollgen ,typeⅢcollgen ,and Ⅰ /Ⅲ collgen ratio in the MI group and the PD group were significantly increased ,the differen had statistical significance (P<0 .01) .The values of the total collgen ,typeⅠcollgen ,typeⅢcollgen ,and Ⅰ /Ⅲ collgen ratio in the PD group were notably decreased than those in the MI group(P<0 .01) .Moreover ,the mRNA and protein levels of NF-kap-paBp65 and MMP-2 in the PD group were lower than those in the MI group ,the difference had statistical significance(P<0 .01) . Conclusion Left ventricular collagen remodeling following acute myocardial infarction could be improved by PDTC to some extent , which mechanism could be related with inhibiting NF-kappaB activation and down -regulating the expression of MMP-2 in rats .

REGγ is a strong candidate for the regulation of cell cycle, proliferation and the invasion by poorly differentiated thyroid carcinoma cells

REGγ is a proteasome activator that facilitates the degradation of small peptides. Abnormally high expression of REGγ has been observed in thyroid carcinomas. The purpose of the present study was to explore the role of REGγ in poorly differentiated thyroid carcinoma (PDTC). For this purpose, small interfering RNA (siRNA) was introduced to down-regulate the level of REGγ in the PDTC cell line SW579. Down-regulation of REGγ at the mRNA and protein levels was confirmed by RT-PCR and Western blot analyses. FACS analysis revealed cell cycle arrest at the G1/S transition, the MTT assay showed inhibition of cell proliferation, and the Transwell assay showed restricted cell invasion. Furthermore, the expression of the p21 protein was increased, the expression of proliferating cell nuclear antigen (PCNA) protein decreased, and the expression of the p27 protein was unchanged as shown by Western blot analyses. REGγ plays a critical role in the cell cycle, proliferation and invasion of SW579 cells. The alteration of p21 and PCNA proteins related to the down-regulation of REGγ suggests that p21 and PCNA participate in the process of REGγ regulation of cell cycle progression and cell proliferation. Thus, targeting REGγ has a therapeutic potential in the management of PDTC patients.

Effect of PDTC on the NF-κB activation and the expression of inflammatory cytokines in THP-1 macrophages induced by rPVL / 中华微生物学和免疫学杂志

Objective To investigate effect of PDTC on the NF-κB activation and the expression of inflammatory cytokines in THP-1 macrophages induced by rPVL.Methods The study was divided into three groups:PBS-treated control group,rPVL-treated group and PDTC group which was given 100 μmol/L PDTC at 60 min before rPVL exposure.Immunohistochemistry method was used to test the translocation of NF-κB protein; the expression of NF-κB and IκB protein was analyzed by Western blot; RT-PCR and ELISA was performed to test expression of IL-8 and L-6 in THP-1 macrophages.Results Compared with rPVL-treated group,the activation of NF-κB and the expression of IL-8 and L-6 in PDTC group was significantly decreased.The protein secretions of IL-8 and IL-6 were reduced to 6.78 ng/ml,3.88 ng/ml,receptively(P ＜0.05).Conclusion The inhibitor of NF-κB,PDTC,could significantly decrease the secretion of pro-inflammatory in THP-1 macrophages by rPVL,and it suggested that PDTC played an important role in protecting tissues from damage induced by rPVL.

Study on the relationship among the NF-?B activation,uPA expression and apoptosis induced by 5- Fu in HCT116 human carcinoma cell line / 肿瘤研究与临床

Objective To study the relationship among apoptosis,NF-KB activation and uPA expres- sion in human colon carcinoma cell line HCTll6 induced by 5-fluorouracil,and to observe the effect of in- hibiting activity of NF-KB by PDTC on apoptosis as well as expression of uPA.Methods Cell apoptosis was analysed by Annexin V-FITC.Fluctuation of NF-KB and uPA was detected by semi-quantitative immuno- histochemistry.Results 5-fluorouracil could induce apoptosis and activate NF-KB.PDTC could significantly increase the apoptosis and suppress the activation of NF-KB induced by 5-fluorouracil.There was a positive correlation between the changes of uPA and NF-KB.Conclusion 5-fluorouracil could induce apoptosis,ac- tivate NF-KB and up-regulate expression of uPA of HCT116 cells.The mechanism of enhanced apoptosis by PDTC may be related to suppressing activation of NF-?B and down-regulating expression of uPA.

The Influence of LPS and PDTC on IL -1? Concentration in Endometriai Cell Culture Supernatant / 医学研究杂志

Objective To learn the influence of density of LPS and LPS + PDTC on the concentration of IL -1? of endometrial cells. Method We used LPS and LPS + PDTC to intervene normal and endometriosis patients, and obtain cultivate supernatant,then detected the concentration density of IL - 1?with ELISA. Result The of IL - 1? had no statistical signifiance in normal endometrial cells, but there was notable change in eutopic and ectopic endometrial cells. Conclusion Activating and refrainning NF -?B through LPS and PDTC, then decteting the change of the ancentration of IL - 1?, understanding the relation between NF -?B and endometriosis are all important in pathogenesis.

Effect on radiation-induced apoptosis of HeLa cells by inhibiting expression of NF-?B / 中国癌症杂志

Background and purpose: NF-?B is activated by tumor necrosis factor,some chemotherapeutic agents,and ionizing radiation.The activation of NF-?B could result in inhibition of apoptosis.NF-?B,regulated by PDTC(pyrrolidine dithiocarbamate),is a specific inhibitor of NF-?B.The purpose of our study was to explore the impact of inhibiting expression of NF-?B on radiation-induced apoptosis of uterine cervix cancer HeLa cells.Methods:Inhibition of NF-?B in HeLa cells was performed by treatment with 100 ?mol/L PDTC for 2 hours.Cells were irradiated at 0,2,4 and 6 Gy with or without PDTC.The expression of NF-?B in nuclear was determined by western blot,and apoptosis was evaluated by using the TUNEL assay.Cell proliferation was evaluated by using MTT assay.Results:NF-?B activation was induced by radiation and inhibited by PDTC.Inhibition of radiation-induced NF-?B activation resulted in inhibiting cell proliferation(P

PDTC reduces E-selectin expression on hypoxia/reoxygenation-stimulated endothelial cells / 中华胸心血管外科杂志

Objective: To study the mechanisms and treatment of ischemia/reperfusion injury. Expression of E selectin was measured and the effect on suppression of E selectin by the Pyrrolidine Dithiocarbamate was investigated. Methods: Endothelial cells were exposed to hypoxia, then returned to reoxygenation condition. ELISA methods were used to detect expression of E selectin. Results: E selectin expression on hypoxia/reoxygenation stimulated endothelial cells increased, PDTC could suppress translation of E selectin effectively. Conclusion: It seems that E selectin may act as a critical factor. PDTC may prove beneficial in the treatment of ischemia/reperfusion injury.

The Protective Effect of Naoyian on Rat Cerebral Microvascular Endothelial Cells Lmpairment Induced by Hydrogen Peroxide / 中国医师杂志

Objective To establish a hydrogen peroxide-impaired model of rat cerebral microvascular endothelial cells in vitro, and observe the protective effect of serum containing Naoyian. Method Rat cerebral microvascular endothelial cells were treated with six concentrations of hydrogen peroxide at four time points. The optimum injury condition of hydrogen peroxide was determined by MTT chromatometry. Then the cultured cells pretreated with serum containing Naoyian and pyrrolidine dithiocarbamate(PDTC) were interfered with hydrogen peroxide in suitable concentration and time, and OD of the treated cells was measured by MTT chromatometry. Results The cultured cells were injured obviously by hydrogen peroxide in 0 125mmol/L for 30min. The OD of the cells pretreated with 5% serum containing Naoyian and 50?mol/L PDTC were significantly higher than that of the cells without pretreatment (P