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AIM: To investigate the mechanism of pyrrolidine dithiocarbamate(PDTC)on transforming growth factor-beta 2(TGF-β2)-induced epithelial-mesenchymal transition(EMT)in human lens epithelial cells(LECs).METHODS: LECs were treated with various doses of PDTC chemicals following TGF-β2 caused EMT on these cells. Cell proliferation and lateral migration were discovered using the CCK-8 and cell scratch test. The markers of EMT, including E-cadherin, α-SMA and nuclear factor-κB(NF-κB)signaling pathway-related expression, were tested by Western Blot as well as the changes in the expression of the apoptosis-related proteins BAX, BCL-2, Caspase-3, and Cyclin D1.RESULTS: The proliferation and migration viability of cells in the TGF-β2 treated group was increased compared to the group without TGF-β2, and the expression of α-SMA increased whereas the E-cadherin expression decreased. With the effect of TGF-β2, NF-κB p65 and phosphorylated NF-κB p65 expression increased, the concentration of TGF-β2 that had the greatest capacity for proliferation and migration was 10 ng/mL(P<0.05). Mechanism study of PDTC-induced EMT reversal and apoptosis showed that cell viability and migratory capability were both significantly reduced after PDTC intervention; PDTC prevents IκB phosphorylation, thus inhibiting NF-κB nuclear translocation. Protein associated to the NF-κB signaling pathway, and protein expression of NF-κB/IκBα/p-IκBα/Iκκ-α/p-Iκκ-α was decreased(P<0.05), PDTC increased the expression of the pro-apoptotic protein BAX/Caspase-3, expression of the inhibitor of apoptosis protein BCL-2 and the cell cycle protein Cyclin D1 was reduced. The expression of NF-κB/IκB mRNA was reduced, expression of the apoptosis-related mRNA BAX increased, while BCL-2 reduced.CONCLUSION: The EMT in LECs cells induced by TGF-β2 can be significantly reversed by PDTC, which may be related to the decreased expression of NF-κB p65/IκB/Iκκ-α and activation of apoptosis-related protein. PDTC can reverse EMT by inhibiting NF-κB signaling pathway and induce apoptosis of abnormally proliferated cells, which will provide new potential therapeutic agents for posterior capsular opacification(PCO)treatment.
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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.
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To evaluate the antioxidant, immunomodulatory and anti-inflammatory activities of pyrrolidine dithiocarbamate and saxagliptin in rats with thioacetamide-induced ulcerative colitis. Methods: Animals were orally administered with a vehicle, sulfasalazine (500 mg/kg), pyrrolidine dithiocarbamate (100 mg/kg), and saxagliptin (10 mg/kg) for two weeks. Ulcerative colitis was induced by a single intrarectal instillation of thioacetamide on day 8. Colon samples were collected to assess mitogen-activated protein kinase (MAPK), phosphorylated extracellular signal-regulated kinase (ERK), cAMP response element-binding protein (CREB), interleukin-12 (IL-12), caspase-3, β-defensin, inducible nitric oxide synthase (iNOS) and glucagon like peptide-1 (GLP-1). Moreover, histopathological examination was performed. Results: Rats treated with thioacetamide caused increases in colonic MAPK, phosphorylated ERK, CREB, caspase-3, IL-12, β-defensin, iNOS, as well as decreases in body weight and GLP-1. In addition, distortion of colonic structure was found by histopathological examination. Pyrrolidine dithiocarbamate and saxagliptin mitigated colitis severity by improving body weight decrease and GLP-1, and reducing colonic MAPK, phosphorylated ERK, CREB, caspase-3, IL-12, β-defensin and iNOS.Conclusions: Pyrrolidine dithiocarbamate and saxagliptin are efficient against thioacetamide induced colitis through improving inflammatory and oxidative changes.
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Objective: To evaluate the antioxidant, immunomodulatory and anti-inflammatory activities of pyrrolidine dithiocarbamate and saxagliptin in rats with thioacetamide-induced ulcerative colitis. Methods: Animals were orally administered with a vehicle, sulfasalazine (500 mg/kg), pyrrolidine dithiocarbamate (100 mg/kg), and saxagliptin (10 mg/kg) for two weeks. Ulcerative colitis was induced by a single intrarectal instillation of thioacetamide on day 8. Colon samples were collected to assess mitogen-activated protein kinase (MAPK), phosphorylated extracellular signal-regulated kinase (ERK), cAMP response element-binding protein (CREB), interleukin-12 (IL-12), caspase-3, β-defensin, inducible nitric oxide synthase (iNOS) and glucagon like peptide-1 (GLP-1). Moreover, histopathological examination was performed. Results: Rats treated with thioacetamide caused increases in colonic MAPK, phosphorylated ERK, CREB, caspase-3, IL-12, β-defensin, iNOS, as well as decreases in body weight and GLP-1. In addition, distortion of colonic structure was found by histopathological examination. Pyrrolidine dithiocarbamate and saxagliptin mitigated colitis severity by improving body weight decrease and GLP-1, and reducing colonic MAPK, phosphorylated ERK, CREB, caspase-3, IL-12, β-defensin and iNOS. Conclusions: Pyrrolidine dithiocarbamate and saxagliptin are efficient against thioacetamide induced colitis through improving inflammatory and oxidative changes.
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Objective To investigate whether pyrrolidine dithiocarbamate (PDTC) can attenuate the acute radiation-induced heart damage (RIHD) by inhibiting the activation of NF-κB and its downstream signaling pathways in rat models. Methods Twenty-one male adult Sprague-Dawley (SD) rats were randomly divided into the blank control, irradiation and PDTC plus irradiation groups (n=7 for each group). In the irradiation and PDTC+ irradiation groups,the rats received 6 MV X-ray at a single fraction of 20.0 Gy. In the PDTC+ irradiation group, intraperitonal injection of PDTC was administered at a dose of 120 mg/kg body weight,30 minutes prior to radiation, once daily for 1-14 days. On the 14thday,pathological changes of myocardial tissue were observed. Masson's trichrome staining was performed to calculate the collagen volume fraction (CVF) of myocardial cells. The expression levels of NF-κB family members including p50, p65,HIF-1α,connective tissue growth factor (CTGF) and collagen type 1(COL-1) proteins and mRNA were quantitatively measured by Western blot and quantitative real-time PCR (qPCR). Statistical analysis was conducted by using t-test. Results HE staining demonstrated that compared with the irradiation group, the severity of myocardial edema was alleviated,the infiltration of inflammatory cells was mitigated and the quantity of fibroblasts was reduced in the PDTC+irradiation group. Masson's trichrome staining revealed that PDCT intervention could decrease the deposition of collagen fiber in the interstitial tissues. Semi-quantitative analysis demonstrated that the CVF value in the PDTC+irradiation group was (9.99± 0.32)%, significantly lower compared with (22.05±0.21)% in the irradiation group (P<0.05). Western blot and qRT-PCR demonstrated that the expression levels of p50,p65,and HIF-1αproteins and mRNA in the PDTC+ irradiation group were significantly down-regulated compared with those in the irradiation group (all P<0.05). Compared with the irradiation group,the expression levels of CTGF protein and mRNA tended to decline (all P>0.05),and the expression levels of COL-1 protein and mRNA were equally inclined to decrease (P<0.05 and P>0.05). Conclusion PDTC can alleviate the acute RIHD by suppressing the activation of NF-κB and its downstream HIF-1α transcription.
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Objective@#To investigate the mechanism of lung injury caused by radiation-induced lung injury by observing the change of nuclear factor (NF-κB) and intercellular adhesion moceule-1 (ICAM-1) in rats and the effects of pyrrolidine dithiocarbamate (PDTC) .@*Methods@#80 SD female rats were randomly classified into 4 groups: control group, radiation group, PDTC treatment group and PDTC group.The radiation induced pulmonary injury model was preformed by using 6 MV X-rays to deliver 8 Gy per day for 5 consecutive days with 40 Gy in total to the thorax of each animal.PDTC was given from 3 d before radiation to 30 d after the first radiation. Rats in control group and PDTC group received the same dose of saline. Animals were sacrificed at 8 week and 24 week after radiation, respectively. The lungs were removed and processed for HE and Masson staining, hydroxyproline content measurement, and real-time quantitative reverse transcription-polymerase chain (RT-PCR) ICAM-1mRNA and NF-κB p65mRNA were detected, Statistical analysis were carried out.@*Results@#Compared with radiation group, there was an obvious amelioration in pathological injury of lung tissue in PDTC treatment group. The lung coefficient and the content of Hyp in PDTC treatment group were significantly lower than those in radiation group (t=3.651, 5.293, 2.348 and 4.126, respectively, allP<0.05) , while slightly higher than those in control group. The levels of ICAM-1mRNA and NF-κB p65mRNA were significantly higher in radiation group than that in PDTC treatment group (allP<0.05) , There were no significant differences in these indicators between control group and PDTC group (P>0.05) .@*Conclusion@#The expression of ICAM-1mRNA and NF-κB p65mRNA is increased in rats with radiation-induced lung injury. Suggest that ICAM-1 and NF-κB are a key factor lead to radiation-induced lung injury. PDTC may inhibits NF-κB activity and further significantly deceases expression of ICAM-1, leading to significantly attenuated pulmonary inflammation and fibrosis, which provides a new therapeutic target for the prevention and treatment of radiation-induced lung injury.
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Objective To study the effect of pyrrolidine dithiocarbamate (PDTC) on the anti-tuberculosis drug-induced liver injury and the molecular mechanism. Methods Clean male SD rats were selected as experimental animals and randomly divided into normal group, model group, PDTC group and AG490 group. Animal model of anti-tuberculosis drug-induced liver injury was established by intragastric administration isoniazid + rifampicin. PDTC group received intraperitoneal injection of PDTC, and AG490 group received intraperitoneal injection of AG490. Twenty-eight days after intervention, the rats were executed, and the liver injury indexes, inflammation indexes and oxidative stress indexes in serum as well as JAK2/STAT3 expression, liver injury indexes, inflammation indexes and oxidative stress indexes in liver tissue were determined. Results p-JAK2, p-STAT3, TNF-α, IL-1β, IL-6, ROS, 8-OHdG and MDA expression in liver tissue as well as TBIL, ALT, AST, γ-GT, TNF-α, IL-1β, IL-6, 8-OHdG and MDA levels in serum of model group were significantly higher than those of normal group while p-JAK2, p-STAT3, TNF-α, IL-1β, IL-6, ROS, 8-OHdG and MDA expression in liver tissue as well as TBIL, ALT, AST, γ-GT, TNF-α, IL-1β, IL-6, 8-OHdG and MDA levels in serum of PDTC group and AG490 group were significantly lower than those of model group. Conclusions PDTC can inhibit the inflammation and oxidative stress mediated by JAK2/STAT3 signaling pathway to alleviate the anti-tuberculosis drug-induced liver injury.
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OBJECTIVE@#To study the effect of pyrrolidine dithiocarbamate (PDTC) on the anti-tuberculosis drug-induced liver injury and the molecular mechanism.@*METHODS@#Clean male SD rats were selected as experimental animals and randomly divided into normal group, model group, PDTC group and AG490 group. Animal model of anti-tuberculosis drug-induced liver injury was established by intragastric administration isoniazid + rifampicin. PDTC group received intraperitoneal injection of PDTC, and AG490 group received intraperitoneal injection of AG490. Twenty-eight days after intervention, the rats were executed, and the liver injury indexes, inflammation indexes and oxidative stress indexes in serum as well as JAK2/STAT3 expression, liver injury indexes, inflammation indexes and oxidative stress indexes in liver tissue were determined.@*RESULTS@#p-JAK2, p-STAT3, TNF-α, IL-1β, IL-6, ROS, 8-OHdG and MDA expression in liver tissue as well as TBIL, ALT, AST, γ-GT, TNF-α, IL-1β, IL-6, 8-OHdG and MDA levels in serum of model group were significantly higher than those of normal group while p-JAK2, p-STAT3, TNF-α, IL-1β, IL-6, ROS, 8-OHdG and MDA expression in liver tissue as well as TBIL, ALT, AST, γ-GT, TNF-α, IL-1β, IL-6, 8-OHdG and MDA levels in serum of PDTC group and AG490 group were significantly lower than those of model group.@*CONCLUSIONS@#PDTC can inhibit the inflammation and oxidative stress mediated by JAK2/STAT3 signaling pathway to alleviate the anti-tuberculosis drug-induced liver injury.
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Objective To study the protective effect of pyrrolidine dithiocarbamate (PDTC) on acute irradiated mice.Methods The 6-8 weeks old male ICR mice were randomly divided into five groups:irradiation alone group (IR),positive control group (amifostine WR-2721 250 mg/kg) and PDTC of 30,60 and 90 mg/kg dose groups.Each group had 10 mice and the drug was given at 0.5 h before whole body irradiation.At 30 d post-irradiation of 7.5 Gy 137 Cs γrays,the mice survival were observed.At 8 d post-irradiation of 5.0 Gy 137 Cs γ-rays,the peripheral blood,hematopoietic system and organ indexes were observed to evaluate the radiation protective effect of PDTC.Results PDTC increased the 30-day survival rates and 60 mg/kg dose had the most obvious effect by increase the survival to 60％ (6/10).The survivals of irradiation alone group and the amifostine positive control group was 10％ (1/10) and 70％ (7/10),respectively.Compared with the irradiation alone group,60 mg/kg PDTC group had the significant difference in spleen index,WBC,HGB,PLT,bone marrow nucleated cells and colony forming unit of spleen (t =2.354,4.793,2.342,6.542,2.649,3.982,P ＜ 0.05).Conclusions PDTC is effective in radiation protection with an optimum dose of 60 mg/kg.
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Objective To evaluate the inhibitory effects of NF-κB inhibitor pyrrolidine dithiocar-bamate (PDTC) on NF-kappa B activity and the serum inflammatory mediators in hypertensive-ventricular hypertrophy-congestive heart fail?ure rats. Methods The rat model of hypertension-cardiac hypertrophy-heart failure was made from 42 male Dahl salt sen?sitive rats. Rats were randomly divided into seven groups including group A (normal diet group), group B (high salt diet group), group C (NF-κB inhibition in early stage), group D (NF-κB inhibition in hypertensive stage), group E (NF-κB inhibi?tion in cardiac hypertrophy stage of week 12) and group G (NF-κB inhibition in heart failure stage). There were six rats for each group. Rats were administrated 8%high salt diet and injected PDTC 100 mg/(kg·d)intraperitoneally according to the prescribed time. Changes of blood pressure, left ventricular end diastolic interventricular septal thickness (IVSD), left ven?tricular end diastolic posterior wall thickness (LVPWD), left ventricular end diastolic diameter (LVEDD), systolic left ventric?ular end diastolic diameter (LVESD), left ventricular ejection fraction (LVEF), heart, lung weight/ body weight ratio, NF-kappa B activity, tumor necrosis factor (TNF)-α, interleukin (IL)-6, monocyte chemotactic protein (MCP)-1 and C-reactive protein (CRP) were observed in different treatment time points of PDCT. Results Levels of NF-κB and proinflammatory cy?tokines were reduced after early administration of PDTC, and the cardiac function was also decreased. The longer the treat?ment time, the greater the protective effect on heart. PDTC can effectively control blood pressure, and block left ventricular hypertrophy and left ventricular failure in a certain extent. The effects of PDTC were limited after persistent hypertension, and myocardial hypertrophy formation accompanied by heart failure. Conclusion PDTC plays a role in prevention and treatment of hypertension, left ventricular hypertrophy and congestive heart failure in model rats. Early application of PDTC could obviously maintain the normal cardiac function in rats with heart disease.
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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.
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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.
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Objective Utilizing the characters of pyrrolidine dithiocarbamate known as a specific inhibitor of NF‐κB and imidapril known as a specific inhibitor of angiotensin converting enzyme ,this study was to initially investigate the relationship between NF‐κBp65 and AngⅡ during the process of myocardial ischemia reperfusion injury .Methods Rat models of myocardial ischemia reper‐fusion injury were successfully established .The observation point is 60 min of reperfusion after 30 min(I30 min R60 min ) ischemia ,the DNA activate of NF‐κBp65 was detected by ELISA ;the content of AngⅡ in blood plasma and cardiac were determined by radioim‐munoassay .We pretreated with pyrrolidine dithiocarbamate ,imidapril ,and both of them to observe changes of the same indexs .Re‐sults Compared with I30 min R60 min ,Pyrrolidine dithiocarbamate ,Imidapril and pyrrolidine dithiocarbamate add imidapril could all ob‐viously inhibit NF‐κBp65 activation and depress the content of Ang Ⅱ in blood plasm and cardiac ,the pyrrolidine dithiocarbamate add imidapril was most obviously .Conclusion During the process of myocardial ischemia‐reperfusion injury ,NF‐κBp65 had been activated ,and the content of Ang Ⅱ in blood plasm and cardiac had increased .NF‐κBp65 and AngⅡ could influence and promote each other during myocardial ischemia reperfusion injury .
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Objective To investigate the effect of pyrrolidine dithiocarbamate(PDTC) on monocyte chemotactic protein1 (MCP-1) in rejection of cardiac allograft and its mechanisms.Methods Heterotopic cervical heart transplantation was performed by cuff-technique.The SD rat recipients were randomly divided into 3 groups:AR group (Acute rejection,n =12),both the recipients and donors were without any treatment.CsA group(n =12),the recipients were treated with 10 mg/kg cyclosporine A after transplantation.PDTC group(n =12),the recipients were treated with 100 mg/kg PDTC after transplantation.All the cardiac allografts were harvested at different time post transplantation according to requirements.We studied allograft myocardial fibrosis wih the help of Masson stain,immuno-histo-chemistry and western blot also were used to detect the expression of MCP-1.Results The survival time of the cardiac allografts was significantly longer in PDTC group than in acute rejection group and CsA group(P ＜ 0.01),and myocardial fibrosis of cardiac allografts in PDTC group was significantly decreased (P ＜ 0.01).The IOD in PDTC group was markedly lower than in CsA group (P ＜ 0.01).Conclusion As the inhibitor of NF-κB,PDTC can significantly relieve rejection of cardiac allograft by inhibiting the expression of MCp-1.
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Testicular torsion results with the damage of the testis and it is a surgical emergency. Pyrrolidine dithiocarbamate (PDTC) is a low-molecular-weight antioxidant and potent inhibitor of nuclear factor kappa B (NF-kappaB) activation. In this study, we aimed to investigate the effects of PDTC to testicular torsion-detorsion (T/D) injury. Forty adult male Sprague-Dawley rats were separated into four groups. A sham operation was performed in group I. In group II, torsion is performed 2 hours by 720 degree extravaginally testis. In group III, 4 h reperfusion of the testis was performed after 2 h of testicular torsion. In group IV, after performing the same surgical procedures as in group III, PDTC (100 mg/kg, intravenous's) was administered before 30 min of detorsion. The testes tissue malondialdehyde (MDA), superoxide dismutase (SOD) catalase (CAT) level was evaluated. Histological evaluations were performed after hematoxylin and eosin staining. Testicular tissue MDA levels were the highest in the T/D groups compared with treatment group. Administration of PDTC prevented a further increase in MDA levels. Significant decrease occurred in CAT and SOD levels in treatment group compared with the control group. The rats in the treatment group had normal testicular architecture. The results suggest that PDTC can be a potential protective agent for preventing the biochemical and histological changes related to oxidative stress in testicular injury caused by testis torsion.
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Adult , Animals , Cats , Humans , Male , Rats , Catalase , Emergencies , Eosine Yellowish-(YS) , Hematoxylin , Malondialdehyde , NF-kappa B , Oxidative Stress , Rats, Sprague-Dawley , Reperfusion , Spermatic Cord Torsion , Superoxide Dismutase , TestisABSTRACT
Objective To observe the effect of signal transduction pathway of NF-κB on tubular cell apoptosis in ischemia-reperfusion induced acute kidney injury (AKI) in mice.Methods Eighteen C57B/6 mice were randomly (random number) divided into three groups,namely control group,AKI group,and pyrrolidine dithiocarbamate (PDTC) group.AKI model of mouse was made by occlusion of bilateral renal pedicles with microvascular clamps for 45 minutes,and intraperitoneal injection of PDTC (50 mg/kg) was given immediately after modeling in mice of PDTC group.Forty-eight hours after modeling,kidney pathological changes,serum creatinine (SCr) and blood urea nitrogen (BUN) were examined,and renal tissue NF-κB,TNFR,Bcl-2 and caspase-3 levels were detected by using immunohistochemistry,and tubular cell apoptosis was observed by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL).Results (1) The pathological Pallers score of renal damage,blood urea nitrogen and serum creatinine levels in PDTC group were significantly lower than those in AKI group [(2.83 ± 0.41)vs.(4.50± 0.55),P=0.000; (61.65 ±3.06) mmol/L vs.(77.78 ±5.82)mmol/L,P=0.000and (74.33 ± 9.83) μmol/L vs.(152.00 ± 16.55) μmol/L,P =0.000,respectively].(2) The level of NF-κB in renal tissue homogenates in PDTC group was significantly lower than that in AKI group [(20.33± 2.34) ％ vs.(35.83 ± 3.06) ％,P =0.000].(3) The apoptotic index of renal tubular cells in PDTC group was significantly lower than that in AKI group [(16.67 ± 1.15) ％ vs.(28.00 ±2.01) ％,P =0.001].(4) The levels of caspase-3 and TNFR1 in renal tissue homogenates in PDTC group were significantly lower than those in AKI group [(7.00 ± 1.26) vs.(11.00 ± 1.26),P =0.000 and (5.55 ± 0.82) vs.(9.75 ± 0.76),P =0.000],and Bcl-2 level in PDTC group was significantly higher than that in AKI group [(10.50± 1.38)vs.(1.83 ±0.98),P=0.000].Conclusions NF-κB activates renal tubular cell apoptosis in acute kidney injury induced in mice after ischemia-reperfusion.Blockade of NF-κB signal transduction pathway may lessen the apoptosis of renal tubular cells,leading to renal function less compromised.
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Objective To investigate the effects of fluoxetine and pyrrolidine dithidarbamate (PDTC) on the lipopolysaccharide(LPS)-induced interleukine-6(IL-6) release in cultured rat astrocytes.Methods The purified astrocytes were cultured in 48-well tissue culture plate and classified into control group,LPS group,fiuoxetine group and PDTC group.Control group and LPS group were cultured as usual,and fluoxetine group and PDTC group were cultured with fluoxetine or PDTC at different concentrations for 48 hours,and then LPS group,Fluoxetine group and PDTC group were incubated with 1 ug/ml LPS for 24 hours.Finally,the levels of IL-6 in the cell supernatant were detected by enzymatic linked immunosorbent assay (ELISA) method.Results The level of IL-6 in LPS group ((1975.46 ± 171.54) pg/ml) was significantly higher than that in control group((10633 ± 782.15)pg/ml) (P ＜ 0.01).The levels of IL-6 were (6198.6 ± 379.4) pg/ml,(4973.6 ± 132.5) pg/ml and (4747.9 ±473.9) pg/ml respectively when the concentrations of fluoxetine were 10 μM,20 μM and 40 μM,and (4821.6 ±180.8) pg/ml,(4735.7 ±620.0)pg/ml and (3525.9 ± 240.0)pg/ml respectively when the concentrations of PDTC were 100 μM,150 μM and 200 μM.There was significant difference in the levels of IL-6 between LPS group and fluoxetine group (P ＜ 0.05),as well as between LPS group and fluoxetine group (P ＜ 0.05).Conclusion LPS can induce IL-6 release from astrocytes,while fluoxetine or PDTC at some concentrations can suppress LPS-induced IL-6 release.
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Objective To explore the effects of pyrrolidine dithiocarbamate (PDTC) on the acute lung injury and the activation of Nrf2 pathway after Paraquat (PQ) induced lung injury.Methods Fortyeight adult male SD rats with lung injury induced by PQ were randomly (random number) divided into control group and PDTC group.Three animals were sacrificed at every 1-week interval,7d,14d and 21 days after PQ intoxication,and the lungs of rats were removed for acute lung injury score after HE staining,and for lung fibrosis assessment after Masson staining,and the levels of reduced glutathione (GSH) and malondialdehyde (MDA) in the lung tissue homogenate were assayed and the phosphorylation of Nrf2 (nuclear-E2-related factor 2) was detected by Weston blot.The mean values of detected variables between two groups were compared by t test,and survival curve was tested by Wilcoxon (Gehan) test.Results The intoxication symptoms of rats were obvious,and 4 rats in control group and 9 rats in PDTC group survived until 21days.The survival time of animals in PDTC group was longer than that in control group (Wilcoxon (Gehan) =10.17023,P =0.001).The levels of MDA in control group were higher than those in PDTC group,while the levels of GSH in control group were lower than those in PDTC group.The levels of phosphorylation of Nrf2 in PDTC group were higher than those in control group at 1-week intervals,1-week:(0.32±0.04) vs.(0.23±0.05),P=0.003; 2-week:(0.62±0.06) vs.(0.33±0.03),P＜0.001; 3-week:(0.61 ±0.04) vs.(0.33±0.05),P＜0.001.The acute lung injury (ALI) scores in PDTC group were lower than those in control group,1-week:(5 ± 0.95) vs.(8 ± 1.23),P =0.002 ; 2-week:(9±1.18) vs.(11±1.02),P=0.019; 3-week:(11±1.33) vs.(12±1.42),P=0.002.The percentages of lung fibrosis at 1-week intervals after PQ intoxication were (40.87 ± 7.25) ％,(43.38 ±5.71)％ and (45.91 ± 3.97)％ in control group,and they were higher than those in PDTC group (32.92±2.34)％,(33.45 ±3.04)％ and (35.27 ±3.81)％ in PDTC group,P=0.017,0.001 and 0.001 respectively.Conclusions Attenuation of acute lung injury and lung fibrosis,and prolongation of survival time of SD rats by PDTC were associated with activation of Nrf2 pathway.
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Objective To observe the levels of Ang - 1 and NF-κB in lung tissue and to aseess the severity of ALI induced by phosgene in order to clarify the mechanism of the protective effect of Ang - 1 on phosgene induced ALI.Method Rats were randomly divided into phosgene group and air group.Another rats were randomly (random number) divided into phosgene group,phosgene + PDTC group and air group.Lung tissue was collected to weigh and calculate the wet / dry weight ratio,measure BALF,white blood cell count,total protein and Ang-1 at given time after exposure to phosgene/air and PDTC.The Ang - 1 and NF-κB levels in lung tissue were measured with Western blot and immunohistochemistry.Data were analyzed by using SPSS 16.0 statistical package and comparisons between groups were carried out byusing One-Way ANOVA analysis and LSD -t test,α ＜ 0.05.Results Serum angiopoietin -1 level became lesser within 48 hours after exposure to Phosgene.The severity of ALI became worser with time elapsing.Ccompare with air group,the severity of ALI in phosgene group was worser with time elapsing ( P ＜ 0.05).Compared with phosgene + PDTC group,the serum angiopoietin -1 and arterial oxygen partial pressure in phosgene group were lower ( P ＜ 0.05).The severity of ALI of rats in phosgene group were worser than that in phosgene + PDTC group ( P ＜ 0.05).Serum angiopoietin -1 and partial pressure of oxygen of rats in phosgene group were higher than those in phosgene + PDTC group ( P ＜ 0.05).Immunohistochemistry test showed that the expression of Ang-1 in lung tissue in air group were normal,and Ang-1 in phosgene group were significantly reduced,and Ang-1 in PDTC intervention group was higher than that in phosgene group and lower than that in air group.The above results were confirmed by Western blot test which was consistent with the results of immunohistochemistry test.Similarly,the levels of NF-κB in lung tissue determined by using both Western - blot and immunohistochemistry were consistant,and results of both methods showed that the expression of NF - κB in air group was normal,and it increased in phosgene group,and the expression of NF-κB in phosgene + PDTC group was lower than that in phosgene group.Conclusions The serum level of Ang-1 was decreasing within 48 hours after ALI.Ang-1 was negatively correlated with the sevfity of phosgene induced ALI.Ang-1 likely had an effect on NF-κB signaling pathway,ameliorating the inflammation mediated by cytokines,reducing lung endothelial permeability and in turn lessening the severity of ALI.
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Objective To investigate the time course of nuclear factor-κB(NF-κB) and the effects of pyrrolidine dithiocarbamate (PDTC) on the expressions of NF-κB,tumor necrosis factor-α (TNF-α) and interleukin1β(IL-1β) in hippocampus after seizures.Methods Epilepsy were induced by [PTZ] through Intraperitoneal injection.Western blotting was used to detect NF-κB p65 expression in nucleus at various experiment groups in different time points( 14d,21 d,28d,35d).Moreover,mRNA and protein expressions of TNF-α and IL-1β in different experiment groups in different time points by Real-time quantitative PCR and ELISA analysis.Results The expression of NF-κB p65 began to increase in the nuclear fraction in 14d,kept rising in 28d and returned to 14d level in 35d after epilepsy seizures,At 14d,21d,28d and 35d,the expressions of NF-κB in PDTC groups ( (0.54 ±0.07),(0.65 ± 0.08 ),(0.78 ± 0.10),(0.78 ± 0.10) ) was significantly lower than those in PTZ groups ((1.20 ±0.11),(1.42 ±0.14),(1.88 ±0.16),(1.25 ±0.10)) (P＜0.01).After epilepsy seizures,the expression of TNF-α 、IL-1β mRNA was increased in PTZ groups( ( 1.34 ±0.13,0.81 ± O.17 ),( 1.64 ±0.17,1.56±0.20),(2.03 ±0.16,1.65 ±0.18),(1.40 ±0.10,1.30 ±0.13) ) than those in NS groups(P＜0.01 ) ;and compared with PTZ groups PDTC significantly decrease the mRNA expressions of TNF-α and IL-1 β in PDTC groups( (0.96 ±0.1,0.57 ±0.07),(1.36 ±0.15,1.09 ±0.18),(1.47 ±0.14,1.25 ±0.16),(1.12 ±0.12,O.85 ± 0.12) ) (P ＜ 0.05 ).The expressions of TN F-α,IL-1β protein were similar in hippocampal by ELISA.Conclusion Seizures induces NF-κB nucleus translocation and promotes the expressions of TNF-ot and IL-1 β in hippocampus and pyrrolidine dithiocarbamate suppresses NF-κB associated inflammatory pathway in epileptic rat hippocampus.