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Background Di(2-ethylhexyl)phthalate (DEHP) and dibutyl phthalate (DBP) are representative environmental endocrine disruptors of phthalate esters (PAEs). Some studies have shown that PAEs exposure may have an impact on lipid metabolism. Objective To investigate the effects of DEHP and/or DBP on lipid metabolism in rats and their possible mechanisms of action. Methods Thirty-six weaned healthy SD male rats, 3 weeks old, weighing 50-70 g, were divided into four groups, i.e., a corn oil control group, a DEHP (750 mg·kg−1) group, a DBP (500 mg·kg−1) group, and a DEHP+DBP (750 mg·kg−1+500 mg·kg−1) group. The rats were exposed to DEHP and/or DBP by oral gavage for 8 weeks, and weighed once a week. The rats were anesthetized 24 h after the last dose, and blood was taken from the apical part of the heart. Serum high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), total cholesterol (TC), and triglyceride (TG) were detected. Liver tissues and perigenital adipose tissues were collected, weighed, and one portion of the tissues was fixed in 10% neutral formalin for pathomorphological observation, and another portion was used for mRNA detection of lipid metabolism-related genes such as Janus kinase 3 (JAK3), signal transducer and activator of transcription 5b (STAT5b), and peroxisome proliferator-activated receptor γ (PPARγ). Results During the DEHP and/or DBP exposure period, the rats in all groups were free to eat and drink without death or injury observed. Compared with the control group: The body weight gain in the DEHP+DBP group was lower at all time points from the 2nd week onwards (P<0.05); the liver organ coefficients of the DEHP and the DEHP+DBP groups were higher (P<0.05); the serum LDL-C levels in the DEHP and the DBP groups were higher (P<0.05). Compared with the DEHP+DBP group: The body weight gains in the DEHP group at the 2nd, 4th, 5th, and 8th weeks were higher (P<0.05), and the body weight gains in the DBP group were higher at all time points except the 1st week (P<0.05); the liver organ coefficients in the DEHP group and the DBP group were lower (P<0.05); the serum TG level in the DEHP group was higher(P<0.05), and the serum LDL-C levels in the DEHP and the DBP groups were higher (P<0.05). The pathomorphological results of liver tissues showed that the hepatocytes in the DEHP, DBP, and DEHP+DBP groups were disordered with loss of cord-like arrangement, swelling (suggesting change of cell proliferation), and presented bilirubin pigmentation. The pathomorphological results of rat perigenital adipose tissues showed had irregular alignment, sizes, and arrangement of adipocyte in the DEHP, DBP, and DEHP+DBP groups. The results of rat liver lipid metabolism-related gene mRNA levels showed that the liver JAK3, STAT5b, and PPARγ mRNA levels in the DEHP, DBP, and DEHP+DBP groups were lower than those in the control group (P<0.05); the rat liver PPARγ mRNA levels in the DEHP and DBP groups were lower than those in the DEHP+DBP group (P<0.05). Conclusion DEHP and/or DBP can inhibit the increase of body weight to varying degrees, induce inflammatory damage to liver tissues, and cause abnormal lipid metabolism in rats, and the associated mechanism may be related to inhibiting the activation of JAK3/STAT5b/PPARγ signaling pathway in rat liver tissues.
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Objective:To study the anti-inflammatory effects of low， middle， and high doses of Anchang decoction on ulcerative colitis in SD rats， and also explore the possible mechanism of Anchang decoction in the prevention and treatment of ulcerative colitis through the effect of different doses on miRNA-146a/non-receptor tyrosine protein kinase（JAK）/signal transduction and activator of transcription 3（STAT3）/cytokine signaling protein-3（SOCS-3） signal pathway and its downstream proteins. Method:The experimental rats were divided into control group ， model group ， mesalazine group（1 g·kg<sup>-1</sup>） and Anchang decoction low（6 g·kg<sup>-1</sup>）， middle（12 g·kg<sup>-1</sup>）and high dose groups（24 g·kg<sup>-1</sup>）， with 10 rats in each group. Except for the control group， 2，4，6-trinitrobenzenesulfonic acid （TNBS）/ethanol enema was used in all the other groups to establish a rat model of ulcerative colitis for 14 days respectively. The general changes of the mental state， stool traits， hair and other general conditions of the rats were observed， and score was graded with reference to the disease activity index （DAI） table. The pathological changes of colon tissue of rats in each group were observed by hematoxylin-eosin （HE） staining. The levels of serum tumor necrosis factor-<italic>α</italic>（TNF-<italic>α</italic>）， interleukin-10（IL-10）， interleukin-17（IL-17）， interleukin-1<italic>β</italic>（IL-1<italic>β</italic>）and interleukin-6（IL-6）were detected by enzyme linked immunosorbent assay （ELISA）. The expression levels of JAK2， phosphorylation STAT3 （p-STAT3）， STAT3 and inhibitor of SOCS-3 in colon tissue were detected by Western blot. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the expression of JAK2， p-STAT3， STAT3， SOCS-3 mRNA in rat colon and miRNA-146a in rat plasma. Result:Compared with the normal group， the expression of JAK2， p-STAT3， STAT3 protein and the expression of JAK2， p-STAT3 and STAT3 mRNA in the model group increased （<italic>P</italic><0.05）， and the relative expression of miRNA-146a， SOCS-3 mRNA and SOCS-3 protein decreased in the model group （<italic>P</italic><0.05）. Compared with the model group， the mental state， food intake， coat color， etc. of rats in the administration groups were significantly improved， the DAI score was significantly reduced （<italic>P</italic><0.05）， the colonic ulcer tissues of rats in the administration groups were improved significantly， the expression levels of JAK2， p-STAT3， STAT3 protein and the expression of JAK2， p-STAT3 and STAT3 mRNA in the colon tissue of the administration groups were decreased （<italic>P</italic><0.05）， and the relative expression levels of miRNA-146a， SOCS-3 mRNA and SOCS-3 protein were increased in the administration groups （<italic>P</italic><0.05）. Conclusion:Anchang decoction can alleviate ulcerative colitis and reduce the activation of inflammatory factors by affecting the expression of genes and proteins related to miRNA-146a/JAK/STAT/SOCS-3 signal transduction pathway.
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@#To study the protective effects of Roudoukou-8 San extract on hypoxia/reoxygenation injury of cardiac myocytes and its relationship with tyrosine protein kinase 2(JAK2)/signal transducer and activator 3(STAT3)signaling pathway, hypoxia/reoxygenation injury model of H9c2 cells was built by sodium dithionite(Na2S2O4). The vitality of the cells was tested by CCK-8; the contents of lactate dehydrogenase(LDH), creatine phosphate kinase(CK)and aspartate aminotransferase(AST)in cell culture medium were tested by fully automatic biochemical analyzer; the contents of catalase(CAT), superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)in cells were tested by kits; cell apoptosis degree was tested by hoechst staining. And the protein expressions of JAK2, p-JAK2, STAT3, p-STAT3, Bcl-2 and Bax in myocardial cells of each group were tested by Western blot. Hypoxia for 1 hour and reoxygenation for 3 hours of 2 mmol/L Na2S2O4 caused damage of about 50% H9c2 cells. Compared with the model group, the extracts of Roudoukou-8 San with different concentrations could increase the viability of cells. Besides, contents of CK, LDH and AST in cell culture medium were decreased significantly, while contents of CAT, SOD and GSH-Px were increased significantly. At the same time, the expression of p-JAK2, p-STAT3 and Bcl-2 were significantly increased and that of Bax was significantly decreased. The effects of Roudoukou-8 San extract could bereduced by AG490 blocker. Therefore, Roudoukou-8 San extract possesses protective effects on Na2S2O4 induced-hypoxia/reoxygenation injury of cardiac myocytes through JAK2/STAT3 signaling pathway.
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To observe the effect of Shudihuang on behaviors and expression of BDNF/TrkB and NRG-3 in prefrontal cortex and striatum of attention deficit hyperactivity disorder (ADHD) model rats. Thirty 4-week-old spontaneous hypertension rats (SHR) were randomly divided into model group, methylphenidate hydrochloride (MPH, 2 mg·kg⁻¹·d⁻¹) and Shudihuang group (2.4 g·kg⁻¹·d⁻¹). Another 10 Wistar-Kyoto (WKY) rats were selected as normal control group. The 0.5% CMC-Na solution was administered to model group and WKY rats (2 mL·kg⁻¹·d⁻¹). All of the rats were treated for 4 weeks. The open field test was performed at the 14th and 28th days after gavage, in order to evaluate the spontaneous and impulsive behaviors. Subsequently, gene and protein expressions of BDNF/TrkB and NRG-3 were tested by RT-qPCR and Western blot. Compared with model group, MPH and Shudihuang groups showed significant reduction in total distance, mean velocity and central distance in the open field test (<0.05), and Shudihuang group displayed a shorter central distance than MPH group (<0.05). RT-qPCR and Western blot analysis indicated that expressions of BDNF/TrkB and NRG-3 were lower in prefrontal cortex and striatum of SHR compared with WKY rats. Four weeks later after administration, both Shudihuang and MPH significantly elevated mRNA and protein expressions of BDNF/TrkB and NRG-3 (<0.05).In conclusion, neurodevelopmental disorder mediated by BDNF/TrkB and NRG-3 was closely related with SHR rats' behaviors. Shudihuang may ameliorate the spontaneous and impulsive behaviors by up-regulating the expressions of BDNF/TrkB and NRG-3 and improving growth and maturation of neurons in SHR.
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JAK-3, a member of the Janus kinase family, is a protein tyrosine kinase, which plays an important role in the JAK-STAT signaling pathway. Previous studies showed that regulation of JAK-3's activity plays a crucial role in the treatment of diseases such as rheumatoid arthritis. Many reports have been published with a focus on selective JAK-3 inhibitors, some of which showed excellent JAK-3 selectivity and inhibitory activities. Among the JAK-3 inhibitors reported, tofacitinib has satisfactory therapeutic benefits in the clinical trials, and has been approved for treatment of patients with rheumatoid arthritis. However, some JAK-3 inhibitors exhibited moderate to severe side effects, which need to be controlled by drug improvement. In order to pave the way for improvement of current JAK-3 inhibitors and development of new JAK-3 inhibitors, we provide an outline of the structure of JAK-3 and strategies in development of its inhibitors.
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Objective To evaluate the method of controlled cortical impact(CCI) on long term cognitive disorder after traumatic brain injury(TBI) and to investigate the possible pathological mechanism.Methods Sixty male SD rats were randomly assigned into 3 groups:sham surgery group(n =10),control group (n =10) and CCI group(n =40).CCI application was used to make the bilateral frontal lobe controlled cortical impact model (depth:1.5 mm,velocity =3.5 m/s,dwell time =400 ms).Morris water maze test and Nissl's staining was used to assess the cognitive function and pathological changes after 8 weeks of CCI.The expressions of brain derived neurotrophic factor (BDNF) and tyrosine protein kinase B (TrkB) mRNA in frontal lobe and hippocampus tissue was detected by reverse transcription polymerase chain reaction(RT-PCR).Results The mortality in CCI group was only 12.5％.Morris water maze test results showed the escape latency in CCI group was longer than that in sham surgery and control groups(F =51.784,P ＜ 0.05).Percent of time spend in goal quarter during probe trial in CCI group was significantly less than that in sham surgery and control groups(F =13.468,P ＜ 0.05).Nissl's staining showed frontal lobe had obviously defects; Nissl's bodies of frontal cortex and CA1 region in hippocampus reduced.The expressions of BDNF and TrkB mRNA in frontal lobe and hippocampus were significantly less than those in sham surgery and control groups(P ＜ 0.05).Conclusions The CCI model can be applied for study on long term cognitive disorder after TBI with good stability and repeatability.Using the experimental parameters of CCI can damage the long term cognitive function after TBI in rats,and lead the pathology changes of brain tissue clearly.This may have some relationship with the expressions of BDNF and TrkB mRNA.
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Objective To investigate the effect of chronic and unpredictable pre-gestational stress on the serum cortisol level of the offspring,as well as the expressions of brain-derived neurotrophic factor (BDNF) and tyrosine protein kinase B (TrkB) in hippocampus when they were 2 month old.Methods Adult female SD rats were divided randomly into 2 groups:the control group and the chronic unpredictable stress (CUS) model group.All rats were tested in the open field test and sucrose intake test before and after CUS protocol.All offspring rats were sacrificed when 2-month old.Serum cortisol (COR) levels were determined by using a standard radioimmunoassay kit.The expressions of BDNF and TrkB in hippocampus were studied by immunoreactivity quantitative analysis.Results 1.After CUS procedure,CUS group showed decreasing activities in the open field test and decreasing sucrose consumption in sucrose intake test compared to the control group and before CUS procedure.2.The serum COR levels in the female offspring rats of CUS group (128.9 ± 7.3) μg/L were higher than those of the control group (119.9 ± 9.0) μg/L,as well as the male offspring of CUS group(116.5 ± 10.9) μg/L compared with the control group(105.4 ± 10.4) μg/L,but the body weight and brain weight between the offspring of 2 groups were not statistic significance.3.Immunoreactivity quantitative analysis showed that the gray values of BDNF in the female offspring of CUS group (36.1 ± 8.5) decreased compared with the control group(42.4 ± 6.9),as well as the male offspring of CUS(39.6 ± 8.4)compared with the control group (43.7 ± 6.4).4.The gray values of TrkB in the female offspring of CUS group (47.1 ± 2.9) decreased than the control group(50.2 ± 3.9),as well as the male offspring of CUS (46.5 ± 6.7)compared with control group(50.5 ± 5.4).Conclusion Pre-gestational stress reduced the expressions of BDNF and TrkB in hippocampus of the offspring which may relate to hypothalamic pituitary adrenal.
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Objective To study the expression of tyrosine protein kinase in hepatocellular carcinoma(HCC)and observe the correlation between tyrosine protein kinase and clinical features.Methods A total of 30 cases with HCC were enrolled in this study.ERK,P38,C-jun,JAK2,STAT3 and STAT5 were detected by immunohistochemical method using tissue chip technology.Results The expression of ERK(0.220±0.033),P38(0.174±0.024),C-jun(0.183±0.064),JAK2(0.192±0.044),STAT3(0.197±0.078)and sTAT5(0.181±0.066)in HCC was significantly higher than that(0.065±0.028,0.058±0.028,0.042±0.016,0.070±0.030,0.052±0.024,0.052±0.023)in cirrhosis 1iver tissues(P＜0.01).There was significantly positive correlation of the expression between ERK,C-jun,JAK2,STAT3 and STAT5(P＜0.01 or P＜0.05).But the expression of P38 was negatively correlated with ERK in the HCC tissues(r=-0.404,P＜0.05).JAK2 had significant correlation with tumor differentiation.The expression of J AKz in stage Ⅲ was significantly higher than that in stage Ⅰ and Ⅱ cancer tissues.Conclusion There is important significance of the excessive activation of MAPK and JAK-STAT signaI transduction in hepatocellular carcinoma process.The unbalance of signal transduction might be one of the pathogenesis of tumor progress.
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Objective To compare the effects of different intervals of theta burst stimulation(TBS) on the expression of long-term potentiation(LTP) and to explore whether LTP is tyrosine protein kinase(TPK)-dependent.in basolateral amygdala(BLA).Methods Basolateral amygdala slices of rats were prepared.Field excitatory post-synaptic potentials(field potential,fEPSPs) of BLA were recorded by stimulating the external capsule.Two TBS's were applied to induce LTP in BLA.Each TBS included a brief,high-frequency pulse train(5 pulses at 100 Hz) given at the theta-rhythm(5Hz) for 4 seconds.Experiments compare the effects of different intervals of two TBS's on the expression of LTP in BLA.The role of tyrosine protein kinase(TPK) on LTP was then determined using bath application of TPK inhibitor genistein.Results Two TBS's of 10 seconds interval failed to induce LTP in BLA.However,two TBS's increased to 10 min and 30 min intervals,individually,both types of stimulations enhanced f-EPSPs.The enhanced f-EPSPs lasted for more than 30 min.LTP induced by two TBS's of 10 min and 30 min interval were blocked by the TPK inhibitor genistein.Conclusion Two TBS's of 10 min intervals was better at the induction of LTP in BLA.The activation of TPK was possibly involved in the induction and maintenance of LTP in the amygdala.
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BACKGROUND: Nerve growth factor(NGF), brain derived neurotrophic factor(BDNF), neurotropin 3(NT-3) and neurotropir-4/5 are neurotrophic factors necessary for the development and maintenance of specific neurors. The tyrosine protein kinase(trk) receptors exhibit specificity for differ ent neurotrophins. NGF is the cognate ligand for the trk A receptor, BDNF binds to trk B receptor and NT-3 binds to irk A, trk B and trk C receptors, Since melanoma cells are devived from neural ectoderm, growth factors which affect neuronal tissue may have a role in melanoma biology. OBJECTIVE: The purpose of this study is to demonstrate the presence of trk receptors in rnelanoma cells and observe th effect of K-252a on these melanoma cells growth and differentiation. METHODS: After K252a over a range of 0-200nM was added into their cell lines, we exam ined cell viability of SK 28 and SK 30 cells. We performed this to examine the expression of the trk by flow cytometry and immunoblotting. RESULTS: 1. The incubation of SK 28 cells and SK 30 cells with K 252a resulted in a dose dependent inhibition of cell proliferation. 2. In the flowcytometry, SK 28 cells and SK 30 cells showed a high expression of trk A and trk B, not trk C. 3. Using immunoblottiiig, trk in SK 28 cells and SK 30 cells was not expressed. CONCLUSIONS: These results indicate that the identification of tyrosine protein kinase reeeptors and their inhibitor which affect differentiation and growth of a melanoma may provide an additional therapeutic option for treatment of melanoma.
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Biología , Encéfalo , Línea Celular , Proliferación Celular , Supervivencia Celular , Ectodermo , Citometría de Flujo , Immunoblotting , Péptidos y Proteínas de Señalización Intercelular , Melanoma , Factor de Crecimiento Nervioso , Factores de Crecimiento Nervioso , Neuronas , Proteínas Tirosina Quinasas , Receptor trkB , Sensibilidad y Especificidad , TirosinaRESUMEN
In recent years, the importance of tyrosine phosphorylation in the nervous system of mammalian is gaining recognition. Tyros ine protein kinases exert important modulatory effect on the proliferation, diff erentiation, migration and metabolism-related singal transduction pathways in c ells. In this paper we reviewed the signal cascade process of three different ty rosine protein kinase families, including Trk, Src and Eph tyrosine protein kina se families. Furthermore, we discussed important role and possible mechanisms of these tyrosine protein kinases on the neuron synapse plasticity and learning an d memory process.