To explore the roles of astrocytes in the epileptogenesis, astrocytes and neurons were isolated, purified and cultured in vitro from cerebral cortex of rats. The astrocytes were activated by ciliary neurotrophic facto...To explore the roles of astrocytes in the epileptogenesis, astrocytes and neurons were isolated, purified and cultured in vitro from cerebral cortex of rats. The astrocytes were activated by ciliary neurotrophic factor (CNTF) and astrocytic conditioned medium (ACM) was collected to treat neurons for 4, 8 and 12 h. By using Western blot, the expression of calmodulin dependent protein kinase Ⅱ (CaMKⅡ), inducible nitric oxide synthase (iNOS) and adenylate cyclase (AC) was de- tected in neurons. The results showed that the expression of CaMKⅡ, iNOS and AC was increased significantly in the neurons treated with ACM from 4 h to 12 h (P<0.05), and that of iNOS and AC peaked at 8 h and 12 h respectively. It was suggested that there might be some epileptogenic factors in the ACM and such signal pathways as NOS-NO-cGMP, Ca2+?CaM-CaMKⅡ and AC-cAMP-PKA might take part in the signal transduction of epileptogenesis.展开更多
Objective: Impaired signal transduction is associated with tumorigenesis and progression of various kinds of human cancers. Transforming growth factor (TGF)-beta/Smad and ras-mitogen activated protein kinase (MAPK...Objective: Impaired signal transduction is associated with tumorigenesis and progression of various kinds of human cancers. Transforming growth factor (TGF)-beta/Smad and ras-mitogen activated protein kinase (MAPK) are two major signal transduction pathways for adjusting cell proliferation and differentiation. Little is known about TGF-beta/Smad4 in non-small cell lung cancer (NSCLC). Hereby, we investigated the expression of Smad4 in NSCLC, its correlation with MAPK proteins (including p38, ERK1 and JNK1 proteins) and their clinical significance in NSCLC. Methods: The expressions of Smad4, p38, ERK1 and JNK1 were detected at protein level with Western blotting and immunohistochemistry, at transcription level with RT-PCR. Statistical analysis was performed for the comparisons of expressions of Smad4, p38, ERK1 and JNK1, and their correlation with various clinicopathological parameters and the prognosis of NSCLC. Results: The levels of protein and mRNA expression of Smad4 in lung cancer tissues were significantly lower than in normal tissues (P〈0.05). All these four proteins were associated with TNM staging. There was a strongly negative correlation between p38 and Smad4. Expressions of Smad4, p38 and JNK1, as well as tumor differentiation and staging were significantly correlated with the prognosis of NSCLC by univariate analysis. By multivariate analysis, only Smad4, p38, tumor differentiation and staging were correlated with the prognosis. Taken together, the negative expression of p38 and positive expression of Smad4 were associated with a better prognosis of NSCLC. Conclusion: Smad4 could be of vital importance for the initiation and development of NSCLC. The expression of Smad4 might be inhibited by p38, supporting a cross-talk between main proteins of TGF-beta/Smad and ras-MAPK signal transduction pathways. Smad4 and p38 could be possible prognostic factors for NSCLC.展开更多
目的研究阻断PI3K-p70S6K和Ras-p42/p44MAPK信号通路对血管紧张素Ⅱ(ANGⅡ)诱导的脐静脉内皮细胞(HUVEC)增殖及细胞周期进程的影响。方法ANGⅡ联合不同浓度雷帕霉素刺激体外培养的HUVEC,3H-胸腺嘧啶核苷掺入法和3H-亮氨酸掺入法测定细...目的研究阻断PI3K-p70S6K和Ras-p42/p44MAPK信号通路对血管紧张素Ⅱ(ANGⅡ)诱导的脐静脉内皮细胞(HUVEC)增殖及细胞周期进程的影响。方法ANGⅡ联合不同浓度雷帕霉素刺激体外培养的HUVEC,3H-胸腺嘧啶核苷掺入法和3H-亮氨酸掺入法测定细胞DNA和蛋白质合成,流式细胞术检测细胞周期变化,免疫印迹(Western blot)检测细胞信号蛋白p70S6K (p70 ribosomal protein S6 kinase),ERK2(extracellular signal regulated kinase)及细胞周期蛋白CyclinD1、CyclinA、和CyclinBt表达的变化。结果雷帕霉素抑制ANGⅡ诱导的血管内皮细胞蛋白质和DNA的合成,并呈剂量依赖性,雷帕霉素抑制ANGⅡ诱导的p70S6K和CyclinD1的表达,阻滞细胞于G1期(P<0.01),而不影响ERK2、CyclinA和CyclinBt的表达。结论PI3K-p70S6K信号通路在ANGⅡ诱导的HUVEC增殖及细胞周期进程中起关键作用,雷帕霉素免疫抑制靶点p70S6K可应用于干预血管内皮细胞的增殖。展开更多
OBJECTIVE:To examine the role and decipher the mechanism of Pingchuan formula(平喘方,PCF)in treating allergic asthma.METHODS:The mice were treated with saline,dexamethasone(DXM)and PCF for 1 week after the asthma mode...OBJECTIVE:To examine the role and decipher the mechanism of Pingchuan formula(平喘方,PCF)in treating allergic asthma.METHODS:The mice were treated with saline,dexamethasone(DXM)and PCF for 1 week after the asthma model was established and their respiratory function including respiratory resistance(RI),pulmonary dynamic compliance(Cdyn)and maximum voluntary ventilation(MVV)were measured.In addition,cellular changes in bronchoalveolar lavage fluid(BALF)and pathological changes in lung biopsy as well as the expression level ofα-smooth muscle actin(α-SMA),transforming growth factor-beta1(TGF-β1)in BALF and interleukin-5(IL-5),interleukin-13(IL-13),tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ),nuclear factor-kappa B-p65(NF-κBp65),inhibitor-αof nuclear transcription factorκB(IκBα),p38 mitogen-activated protein kinase(p38 MAPK),c-jun n-terminal kinase(JNK)and its phosphorylated proteins in lung tissue were also examined and compared among different groups.RESULTS:Our data suggested that the respiratory functions were significantly improved and the pathological changes ameliorated in the DXM group and the PCF group compared to the model group.Both DXM and PCF effectively decreased the number of eosinophils,lymphocytes,and neutrophils in BAL as well as the secretion ofα-SMA and TGF-β1,IL-5,IL-13,while increased the expression of TNF-αand IFN-γ.Furthermore,our study indicated that the NF-κBp65,IκBα,p38 MAPK and JNK pathways were inhibited under the treatment of PCF.CONCLUSION:Our data indicated that PCF can attenuate the inflammatory response in asthma through inhibiting the NF-κB/MAPK signaling pathway.This study not only supported the use of PCF in allergic asthma in clinic but also shed light upon afurther understanding of the disease pathogenesis.展开更多
Background Urotensin Ⅱ (Ull),a potent vasoconstrictive peptide,is able to stimulate phenotypic differentiation of adventitial fibroblasts.This study aimed to determine the effect of UII on monocyte chemoattractant ...Background Urotensin Ⅱ (Ull),a potent vasoconstrictive peptide,is able to stimulate phenotypic differentiation of adventitial fibroblasts.This study aimed to determine the effect of UII on monocyte chemoattractant protein-1 (MCP1) expression in rat aortic adventitial fibroblasts,so as to explore possible mechanisms in the development of vascular inflammation.Methods Growth-arrested adventitial fibroblasts were incubated in serum-free medium with UII (1010-10-7 mol/L) and inhibitors of signal transduction pathways for 1 to 24 hours.MCP-1 mRNA and protein expression and secretion were determined by RT-PCR,Western blotting analysis and enzyme-linked immunosorbent assay (ELISA),respectively.Results UII dose-and time-dependently promoted MCP-1 mRNA and protein expression and secretion in cells,with maximal effect at 10-8 mol/L at 3 hours for mRNA expression,24 hours for protein expression in the cells,and 12 hours for protein secretion from the cells.Furthermore,the UII effects were significantly inhibited by treatment with its receptor antagonist SB710411,Rho kinase inhibitor Y27632,protein kinase C (PKC) inhibitor H7,mitogen-activated protein kinase inhibitor PD98059,calcineurin inhibitor cyclosporine A,and the Ca2+channel blocker nicardipine.Conclusion UII may stimulate MCP-1 expression in rat aortic adventitial fibroblasts through its receptor and Rho kinase,PKC,mitogen-activated protein kinase,calcineurin and Ca2+ channel signal transduction,thus contributing to adventitial inflammation.展开更多
Background The role of the Gαq/11-mediated signal transduction pathway in angiotensin Ⅱ (Ang Ⅱ) induced cardiac hypertrophy remains unclear. This study was to investigate the role of the Gαq/11 signal transducti...Background The role of the Gαq/11-mediated signal transduction pathway in angiotensin Ⅱ (Ang Ⅱ) induced cardiac hypertrophy remains unclear. This study was to investigate the role of the Gαq/11 signal transduction pathway in the development of cardiac hypertrophy in 2K1C hypertensive rats and in cultured neonatal rat ventricular myocytes (NRVMs) and to elucidate the effects of the pathway on Ang Ⅱ induced cardiac hypertrophy. Methods Renal hypertension was induced in 2K1C hypertensive rats by placing a silver clip around the left renal artery. At 8 weeks after operation,the systolic blood pressure,the ratio of left ventricular weight to body weight (LV/BW),and the concentration of AngⅡ in the heart were measured. The protein levels of Gαq/11 and extracellular signal-regulated kinase 1/2 (ERK1/2) were assayed by Western blot analysis,and the activity of phospholipase C (PLC) in the myocardium was detected using [ 3H]-PIP_2 as a substrate. Changes in [ 3H]-leucine incorporation and in the protein levels of the signal molecules Gαq/11,PLCβ_3,and ERK1/2 were measured after NRVMs were stimulated with 10 -7 mol/L AngⅡ. Results The protein levels of Gαq/11 and ERK1/2 in the hearts of 2K1C rats increased by 35.8% and 31.9%,respectively,compared with the sham group. The PLC activity in the 2K1C group was also significantly increased ( P <0.05). The levels of Gαq/11,PLCβ_3,and ERK1/2 increased significantly after NRVMs were stimulated by AngⅡ. The upregulation of Gαq/11,PLCβ_3 and ERK1/2 in NRVMs occurred prior to [ 3H]-leucine incorporation increases,and could be inhibited with losartan. Conclusion AngⅡ can initiate cardiac hypertrophy and upregulate signal molecules in the Gαq/11-mediated signal transduction pathway,such as Gαq/11,PLCβ_3 and ERK1/2,at both tissue and cellular levels.展开更多
OBJECTIVE:To investigate the efficacy of needling acupoins of Guanyuan(CV4),Sanyinjiao(SP6),Zusanli(ST36),Pishu(BL20),Shenshu(BL23),Zigong(EX-CA1)on the expression of p38 mitogen-activated protein kinase(p38MAPK)in ov...OBJECTIVE:To investigate the efficacy of needling acupoins of Guanyuan(CV4),Sanyinjiao(SP6),Zusanli(ST36),Pishu(BL20),Shenshu(BL23),Zigong(EX-CA1)on the expression of p38 mitogen-activated protein kinase(p38MAPK)in ovarian tissue in rats with premature ovarian failure induced by cyclophosphamide,and to study the underlying mechanism.METHODS:Forty specific pathogen free female Sprague-Dawley rats were randomly divided into the blank group,the model group,the acupuncture group,the Western Medicine group and the Western Medicine combined with acupuncture group.Except the blank group,the rest of the rats were given with cyclophosphamide for 14 d to establish premature ovarian failure model.No intervention was conducted in the blank group and model group;the acupuncture group was given with acupuncture daily;the Western Medicine group was given with estradiol valerate(0.09 mg/kg)by intragastrical gavage daily;the combination group was given with acupuncture combined with estradiol valerate(0.09 mg/kg)daily.Each group was intervened in continuously for 14 d.After the last treatment,the levels of estradiol(E2),follicle stimulating hormone(FSH)and luteinizing hormone(LH)were detected by enzyme-linked immunosorbent assay,then the ovarian tissue was dissected.Western blot was used to detect the expression of p38 MAPK protein.RESULTS:Compared with the blank group,E2 in the serum of rats in the model group was significantly decreased(P<0.05),FSH and LH were significantly increased(P<0.05),and the expression of p38 MAPK protein in the ovarian tissue of the rats was significantly increased(P<0.05).Compared with the model group,E2 in the serum of the acupuncture group,Western Medicine group and the combination group were significantly increased(P<0.05),FSH and LH levels were significantly decreased(P<0.05),and the expression of p38MAPK protein in the ovarian tissue of the rats was significantly decreased(P<0.05).However,there was no statistically significant difference between the Western Medicine group and the acupuncture group(P>0.05).CONCLUSIONS:Acupuncture has the same effect as estrogen in interfering POF caused by cyclophosphamide,and its mechanism may be related to inhibiting the expression of p38MAPK protein in ovarian tissue and affecting the activation of p38MAPK signaling pathway.展开更多
The aim of this study was to investigate the role of p38 mitogen-activated protein kinase(MAPK)in cell migration induced by platelet-derived growth factor(PDGF).Western blot was performed to detect the phosphorylation...The aim of this study was to investigate the role of p38 mitogen-activated protein kinase(MAPK)in cell migration induced by platelet-derived growth factor(PDGF).Western blot was performed to detect the phosphorylation of p38 in NIH3T3 cells treated with PDGF.A Transwell cell migration system was used to determine the effects of PDGF treatment on the migration of NIH3T3 cells and the influence of p38 deficiency on this process in a p38 gene knockout(p38^(−/−))mouse embryonic fibroblast cell line.On the stimulation of PDGF,the migration of NIH3T3 cells was significantly increased(P<0.001)compared to the control and p38 MAP kinase was simultaneously phosphorylated.Furthermore,the PDGF-induced cell migration was significantly blocked in p38 gene knockout(p38^(−/−))mouse embryonic fibroblasts(MEFs)(P<0.001)as compared with the wild type cells(p38+/+).p38 MAPK plays an important role in the regulation of cell migration induced by PDGF.展开更多
OBJECTIVES: To study the expression of inflammatory signal in local prostate tissue of chronic pelvic pain syndrome(CPPS) rats by electroacupuncture(EA) of Guanyuan(CV4), Zhongji(CV3), Huiyang(BL35) and Sanyinjiao(SP6...OBJECTIVES: To study the expression of inflammatory signal in local prostate tissue of chronic pelvic pain syndrome(CPPS) rats by electroacupuncture(EA) of Guanyuan(CV4), Zhongji(CV3), Huiyang(BL35) and Sanyinjiao(SP6), and to explore the possible mechanism of anti-inflammatory and analgesic effects of EA. METHODS : A total of 36 Sprague-Dawley male rats were randomly divided into three groups: control, model and EA(n=12 rats/group). The CPPS model was made by injection of CFA into ventral lobes of the prostate(0.1 m L). Electric acupuncture apparatus was applied to stimulate Guanyuan(CV4), Zhongji(CV3), bilateral Huiyang(BL35) and Sanyinjiao(SP6) acupoints in EA group. The general condition of rats was observed and the prostate index(PI) was calculated. The thermal pain threshold was collected after each therapeutic course. Histopathological changes of the prostate tissue were examined by hematoxylin-eosin staining method. The expression levels of tumor necrosis factor α(TNF-α), interleukin-1β(IL-1β) and prostaglandin E2(PGE2) in prostatic homogenates were measured by enzyme linked immunosorbent assay(ELISA). Moreover, the expression levels of purinergic 2X7 receptor(P2X7R), NOD-like receptor pyrin domain-containing 3(NLRP3), caspase-1 and interleukin-18(IL-18) m RNA were quantified by quantitative real-time polymerase chain reaction. RESULTS: Compared with control group, the PI of rats increased, and the thermal pain threshold decreased significantly in model group. The morphological structure of prostate tissues of rats in model group was severely damaged with a large number of inflammatory cells infiltration. Additionally, the levels of TNF-α, IL-1β and PGE2 were higher, and the expressions of P2X7R, NLRP3, caspase-1 and IL-18 m RNA were higher than those in control group. After EA treatment, the PI was significantly decreased, the thermal pain threshold was significantly increased, and the tissue damage was significantly improved. The expressions of inflammatory cytokines were lower in EA group, and expression of P2X7R/NLRP3 pathway was down-regulated. CONCLUSION: The effect of EA at Guanyuan(CV4), Zhongji(CV3), Huiyang(BL35) and Sanyinjiao(SP6) can improve inflammation and pain symptoms of CPPS rats induced by Complete Freund’s adjuvant(CFA). This suggests that EA at Guanyuan(CV4), Zhongji(CV3), Huiyang(BL35) and Sanyinjiao(SP6) can produce antiinflammatory analgesia effect by preventing the activation of P2X7R/NLRP3 signal pathway, inhibit the release of inflammatory cytokines in CPPS rats, which may provide a putative novel target for the treatment of CPPS.展开更多
基金a grant from Doctor Point Foundation of Ministry of Education of China (No. 2004 0487060)
文摘To explore the roles of astrocytes in the epileptogenesis, astrocytes and neurons were isolated, purified and cultured in vitro from cerebral cortex of rats. The astrocytes were activated by ciliary neurotrophic factor (CNTF) and astrocytic conditioned medium (ACM) was collected to treat neurons for 4, 8 and 12 h. By using Western blot, the expression of calmodulin dependent protein kinase Ⅱ (CaMKⅡ), inducible nitric oxide synthase (iNOS) and adenylate cyclase (AC) was de- tected in neurons. The results showed that the expression of CaMKⅡ, iNOS and AC was increased significantly in the neurons treated with ACM from 4 h to 12 h (P<0.05), and that of iNOS and AC peaked at 8 h and 12 h respectively. It was suggested that there might be some epileptogenic factors in the ACM and such signal pathways as NOS-NO-cGMP, Ca2+?CaM-CaMKⅡ and AC-cAMP-PKA might take part in the signal transduction of epileptogenesis.
基金This work was supported by the National Natural Science Foundation of China(No.30100220)
文摘Objective: Impaired signal transduction is associated with tumorigenesis and progression of various kinds of human cancers. Transforming growth factor (TGF)-beta/Smad and ras-mitogen activated protein kinase (MAPK) are two major signal transduction pathways for adjusting cell proliferation and differentiation. Little is known about TGF-beta/Smad4 in non-small cell lung cancer (NSCLC). Hereby, we investigated the expression of Smad4 in NSCLC, its correlation with MAPK proteins (including p38, ERK1 and JNK1 proteins) and their clinical significance in NSCLC. Methods: The expressions of Smad4, p38, ERK1 and JNK1 were detected at protein level with Western blotting and immunohistochemistry, at transcription level with RT-PCR. Statistical analysis was performed for the comparisons of expressions of Smad4, p38, ERK1 and JNK1, and their correlation with various clinicopathological parameters and the prognosis of NSCLC. Results: The levels of protein and mRNA expression of Smad4 in lung cancer tissues were significantly lower than in normal tissues (P〈0.05). All these four proteins were associated with TNM staging. There was a strongly negative correlation between p38 and Smad4. Expressions of Smad4, p38 and JNK1, as well as tumor differentiation and staging were significantly correlated with the prognosis of NSCLC by univariate analysis. By multivariate analysis, only Smad4, p38, tumor differentiation and staging were correlated with the prognosis. Taken together, the negative expression of p38 and positive expression of Smad4 were associated with a better prognosis of NSCLC. Conclusion: Smad4 could be of vital importance for the initiation and development of NSCLC. The expression of Smad4 might be inhibited by p38, supporting a cross-talk between main proteins of TGF-beta/Smad and ras-MAPK signal transduction pathways. Smad4 and p38 could be possible prognostic factors for NSCLC.
文摘目的研究阻断PI3K-p70S6K和Ras-p42/p44MAPK信号通路对血管紧张素Ⅱ(ANGⅡ)诱导的脐静脉内皮细胞(HUVEC)增殖及细胞周期进程的影响。方法ANGⅡ联合不同浓度雷帕霉素刺激体外培养的HUVEC,3H-胸腺嘧啶核苷掺入法和3H-亮氨酸掺入法测定细胞DNA和蛋白质合成,流式细胞术检测细胞周期变化,免疫印迹(Western blot)检测细胞信号蛋白p70S6K (p70 ribosomal protein S6 kinase),ERK2(extracellular signal regulated kinase)及细胞周期蛋白CyclinD1、CyclinA、和CyclinBt表达的变化。结果雷帕霉素抑制ANGⅡ诱导的血管内皮细胞蛋白质和DNA的合成,并呈剂量依赖性,雷帕霉素抑制ANGⅡ诱导的p70S6K和CyclinD1的表达,阻滞细胞于G1期(P<0.01),而不影响ERK2、CyclinA和CyclinBt的表达。结论PI3K-p70S6K信号通路在ANGⅡ诱导的HUVEC增殖及细胞周期进程中起关键作用,雷帕霉素免疫抑制靶点p70S6K可应用于干预血管内皮细胞的增殖。
基金Supported by Natural Science Foundation-funded Project:Pingchuan Formula Regulates the Effect of Pi3k/Akt Pathway on Airway Inflammation and Airway Remodeling in Asthma and its Mechanism(No.81603662)Shanghai Municipal Commission of Health and Family Planning:"Shanghai School of Traditional Chinese Medicine"Xu’s Pediatric Diagnosis and Treatment[Center Construction ZY(2018-2020)-CCCX-1012]。
文摘OBJECTIVE:To examine the role and decipher the mechanism of Pingchuan formula(平喘方,PCF)in treating allergic asthma.METHODS:The mice were treated with saline,dexamethasone(DXM)and PCF for 1 week after the asthma model was established and their respiratory function including respiratory resistance(RI),pulmonary dynamic compliance(Cdyn)and maximum voluntary ventilation(MVV)were measured.In addition,cellular changes in bronchoalveolar lavage fluid(BALF)and pathological changes in lung biopsy as well as the expression level ofα-smooth muscle actin(α-SMA),transforming growth factor-beta1(TGF-β1)in BALF and interleukin-5(IL-5),interleukin-13(IL-13),tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ),nuclear factor-kappa B-p65(NF-κBp65),inhibitor-αof nuclear transcription factorκB(IκBα),p38 mitogen-activated protein kinase(p38 MAPK),c-jun n-terminal kinase(JNK)and its phosphorylated proteins in lung tissue were also examined and compared among different groups.RESULTS:Our data suggested that the respiratory functions were significantly improved and the pathological changes ameliorated in the DXM group and the PCF group compared to the model group.Both DXM and PCF effectively decreased the number of eosinophils,lymphocytes,and neutrophils in BAL as well as the secretion ofα-SMA and TGF-β1,IL-5,IL-13,while increased the expression of TNF-αand IFN-γ.Furthermore,our study indicated that the NF-κBp65,IκBα,p38 MAPK and JNK pathways were inhibited under the treatment of PCF.CONCLUSION:Our data indicated that PCF can attenuate the inflammatory response in asthma through inhibiting the NF-κB/MAPK signaling pathway.This study not only supported the use of PCF in allergic asthma in clinic but also shed light upon afurther understanding of the disease pathogenesis.
基金This project was supported by grants from National Natural Science Foundation of China (No. 30971273 and No. 81270223) and Natural Science Foundation of Guangdong Province of China (No. 9151051501000016 and No. S2011010000450).
文摘Background Urotensin Ⅱ (Ull),a potent vasoconstrictive peptide,is able to stimulate phenotypic differentiation of adventitial fibroblasts.This study aimed to determine the effect of UII on monocyte chemoattractant protein-1 (MCP1) expression in rat aortic adventitial fibroblasts,so as to explore possible mechanisms in the development of vascular inflammation.Methods Growth-arrested adventitial fibroblasts were incubated in serum-free medium with UII (1010-10-7 mol/L) and inhibitors of signal transduction pathways for 1 to 24 hours.MCP-1 mRNA and protein expression and secretion were determined by RT-PCR,Western blotting analysis and enzyme-linked immunosorbent assay (ELISA),respectively.Results UII dose-and time-dependently promoted MCP-1 mRNA and protein expression and secretion in cells,with maximal effect at 10-8 mol/L at 3 hours for mRNA expression,24 hours for protein expression in the cells,and 12 hours for protein secretion from the cells.Furthermore,the UII effects were significantly inhibited by treatment with its receptor antagonist SB710411,Rho kinase inhibitor Y27632,protein kinase C (PKC) inhibitor H7,mitogen-activated protein kinase inhibitor PD98059,calcineurin inhibitor cyclosporine A,and the Ca2+channel blocker nicardipine.Conclusion UII may stimulate MCP-1 expression in rat aortic adventitial fibroblasts through its receptor and Rho kinase,PKC,mitogen-activated protein kinase,calcineurin and Ca2+ channel signal transduction,thus contributing to adventitial inflammation.
文摘Background The role of the Gαq/11-mediated signal transduction pathway in angiotensin Ⅱ (Ang Ⅱ) induced cardiac hypertrophy remains unclear. This study was to investigate the role of the Gαq/11 signal transduction pathway in the development of cardiac hypertrophy in 2K1C hypertensive rats and in cultured neonatal rat ventricular myocytes (NRVMs) and to elucidate the effects of the pathway on Ang Ⅱ induced cardiac hypertrophy. Methods Renal hypertension was induced in 2K1C hypertensive rats by placing a silver clip around the left renal artery. At 8 weeks after operation,the systolic blood pressure,the ratio of left ventricular weight to body weight (LV/BW),and the concentration of AngⅡ in the heart were measured. The protein levels of Gαq/11 and extracellular signal-regulated kinase 1/2 (ERK1/2) were assayed by Western blot analysis,and the activity of phospholipase C (PLC) in the myocardium was detected using [ 3H]-PIP_2 as a substrate. Changes in [ 3H]-leucine incorporation and in the protein levels of the signal molecules Gαq/11,PLCβ_3,and ERK1/2 were measured after NRVMs were stimulated with 10 -7 mol/L AngⅡ. Results The protein levels of Gαq/11 and ERK1/2 in the hearts of 2K1C rats increased by 35.8% and 31.9%,respectively,compared with the sham group. The PLC activity in the 2K1C group was also significantly increased ( P <0.05). The levels of Gαq/11,PLCβ_3,and ERK1/2 increased significantly after NRVMs were stimulated by AngⅡ. The upregulation of Gαq/11,PLCβ_3 and ERK1/2 in NRVMs occurred prior to [ 3H]-leucine incorporation increases,and could be inhibited with losartan. Conclusion AngⅡ can initiate cardiac hypertrophy and upregulate signal molecules in the Gαq/11-mediated signal transduction pathway,such as Gαq/11,PLCβ_3 and ERK1/2,at both tissue and cellular levels.
文摘OBJECTIVE:To investigate the efficacy of needling acupoins of Guanyuan(CV4),Sanyinjiao(SP6),Zusanli(ST36),Pishu(BL20),Shenshu(BL23),Zigong(EX-CA1)on the expression of p38 mitogen-activated protein kinase(p38MAPK)in ovarian tissue in rats with premature ovarian failure induced by cyclophosphamide,and to study the underlying mechanism.METHODS:Forty specific pathogen free female Sprague-Dawley rats were randomly divided into the blank group,the model group,the acupuncture group,the Western Medicine group and the Western Medicine combined with acupuncture group.Except the blank group,the rest of the rats were given with cyclophosphamide for 14 d to establish premature ovarian failure model.No intervention was conducted in the blank group and model group;the acupuncture group was given with acupuncture daily;the Western Medicine group was given with estradiol valerate(0.09 mg/kg)by intragastrical gavage daily;the combination group was given with acupuncture combined with estradiol valerate(0.09 mg/kg)daily.Each group was intervened in continuously for 14 d.After the last treatment,the levels of estradiol(E2),follicle stimulating hormone(FSH)and luteinizing hormone(LH)were detected by enzyme-linked immunosorbent assay,then the ovarian tissue was dissected.Western blot was used to detect the expression of p38 MAPK protein.RESULTS:Compared with the blank group,E2 in the serum of rats in the model group was significantly decreased(P<0.05),FSH and LH were significantly increased(P<0.05),and the expression of p38 MAPK protein in the ovarian tissue of the rats was significantly increased(P<0.05).Compared with the model group,E2 in the serum of the acupuncture group,Western Medicine group and the combination group were significantly increased(P<0.05),FSH and LH levels were significantly decreased(P<0.05),and the expression of p38MAPK protein in the ovarian tissue of the rats was significantly decreased(P<0.05).However,there was no statistically significant difference between the Western Medicine group and the acupuncture group(P>0.05).CONCLUSIONS:Acupuncture has the same effect as estrogen in interfering POF caused by cyclophosphamide,and its mechanism may be related to inhibiting the expression of p38MAPK protein in ovarian tissue and affecting the activation of p38MAPK signaling pathway.
基金This work was supported by the National Basic Research Program of China(973 Program)(No.2002CB513005)the National Natural Science Foundation of China(Grant No.30572151)+1 种基金Guangdong Provincial Science and Technology Program(No.A1090202)the Medical Science and Technology Foundation of Guangdong Province(No.A2005367).
文摘The aim of this study was to investigate the role of p38 mitogen-activated protein kinase(MAPK)in cell migration induced by platelet-derived growth factor(PDGF).Western blot was performed to detect the phosphorylation of p38 in NIH3T3 cells treated with PDGF.A Transwell cell migration system was used to determine the effects of PDGF treatment on the migration of NIH3T3 cells and the influence of p38 deficiency on this process in a p38 gene knockout(p38^(−/−))mouse embryonic fibroblast cell line.On the stimulation of PDGF,the migration of NIH3T3 cells was significantly increased(P<0.001)compared to the control and p38 MAP kinase was simultaneously phosphorylated.Furthermore,the PDGF-induced cell migration was significantly blocked in p38 gene knockout(p38^(−/−))mouse embryonic fibroblasts(MEFs)(P<0.001)as compared with the wild type cells(p38+/+).p38 MAPK plays an important role in the regulation of cell migration induced by PDGF.
基金Supported by Fundamental Research Funds for the Central Universities Project:RNA Sequencing Technology Screening the Mechanism of Acupuncture on Pain in Chronic Pelvic Pain Syndrome Rats (2020-JYB-XJSJJ-018)。
文摘OBJECTIVES: To study the expression of inflammatory signal in local prostate tissue of chronic pelvic pain syndrome(CPPS) rats by electroacupuncture(EA) of Guanyuan(CV4), Zhongji(CV3), Huiyang(BL35) and Sanyinjiao(SP6), and to explore the possible mechanism of anti-inflammatory and analgesic effects of EA. METHODS : A total of 36 Sprague-Dawley male rats were randomly divided into three groups: control, model and EA(n=12 rats/group). The CPPS model was made by injection of CFA into ventral lobes of the prostate(0.1 m L). Electric acupuncture apparatus was applied to stimulate Guanyuan(CV4), Zhongji(CV3), bilateral Huiyang(BL35) and Sanyinjiao(SP6) acupoints in EA group. The general condition of rats was observed and the prostate index(PI) was calculated. The thermal pain threshold was collected after each therapeutic course. Histopathological changes of the prostate tissue were examined by hematoxylin-eosin staining method. The expression levels of tumor necrosis factor α(TNF-α), interleukin-1β(IL-1β) and prostaglandin E2(PGE2) in prostatic homogenates were measured by enzyme linked immunosorbent assay(ELISA). Moreover, the expression levels of purinergic 2X7 receptor(P2X7R), NOD-like receptor pyrin domain-containing 3(NLRP3), caspase-1 and interleukin-18(IL-18) m RNA were quantified by quantitative real-time polymerase chain reaction. RESULTS: Compared with control group, the PI of rats increased, and the thermal pain threshold decreased significantly in model group. The morphological structure of prostate tissues of rats in model group was severely damaged with a large number of inflammatory cells infiltration. Additionally, the levels of TNF-α, IL-1β and PGE2 were higher, and the expressions of P2X7R, NLRP3, caspase-1 and IL-18 m RNA were higher than those in control group. After EA treatment, the PI was significantly decreased, the thermal pain threshold was significantly increased, and the tissue damage was significantly improved. The expressions of inflammatory cytokines were lower in EA group, and expression of P2X7R/NLRP3 pathway was down-regulated. CONCLUSION: The effect of EA at Guanyuan(CV4), Zhongji(CV3), Huiyang(BL35) and Sanyinjiao(SP6) can improve inflammation and pain symptoms of CPPS rats induced by Complete Freund’s adjuvant(CFA). This suggests that EA at Guanyuan(CV4), Zhongji(CV3), Huiyang(BL35) and Sanyinjiao(SP6) can produce antiinflammatory analgesia effect by preventing the activation of P2X7R/NLRP3 signal pathway, inhibit the release of inflammatory cytokines in CPPS rats, which may provide a putative novel target for the treatment of CPPS.