AIM: To investigate whether the regulation of aquaporin 3 (AQP3) and AQP9 induced by Auphen and dibutyryl cAMP (dbcAMP) inhibits hepatic tumorigenesis.METHODS: Expression of AQP3 and AQP9 was detected by Western blot,...AIM: To investigate whether the regulation of aquaporin 3 (AQP3) and AQP9 induced by Auphen and dibutyryl cAMP (dbcAMP) inhibits hepatic tumorigenesis.METHODS: Expression of AQP3 and AQP9 was detected by Western blot, immunohistochemistry (IHC), and RT-PCR in HCC samples and paired non-cancerous liver tissue samples from 30 hepatocellular carcinoma (HCC) patients. A xenograft tumor model was used in vivo. Nine nude mice were divided into control, Auphen-treated, and dbcAMP-treated groups (n = 3 for each group). AQP3 and AQP9 protein expression after induction of xenograft tumors was detected by IHC and mRNA by RT-PCR analysis. The terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay and histological evaluation were used to detect apoptosis of tumor cells, and the concentration of serum α-fetoprotein (AFP) was measured using RT-PCR and an ELISA kit.RESULTS: The volumes and weights of tumors decreased significantly in the Auphen- and dbcAMP-treated mice compared with the control mice (P < 0.01). The levels of AQP3 were significantly lower in the Auphen treatment group, and levels of AQP9 were significantly higher in thedbcAMP treatment mice than in the control mice (P < 0.01). The reduction of AQP3 by Auphen and increase of AQP9 by dbcAMP in nude mice suppressed tumor growth of HCC, which resulted in reduced AFP levels in serum and tissues, and apoptosis of tumor cells in the Auphen- and dbcAMP-treated mice, when compared with control mice (P < 0.01). Compared with para-carcinoma tissues, AQP3 expression increased in tumor tissues whereas the expression of AQP9 decreased. By correlating clinicopathological and expression levels, we demonstrated that the expression of AQP3 and AQP9 was correlated with clinical progression of HCC and disease outcomes.CONCLUSION: AQP3 increases in HCC while AQP9 decreases. Regulation of AQP3 and AQP9 expression by Auphen and dbcAMP inhibits the development and growth of HCC.展开更多
Aim: To study the influences of dibutyryl cyclic adenosine monophosphate (dbcAMP) and forskolin on human sperm motility in vitro. Methods: Semen samples, aseptically obtained by masturbation and prepared by swim-up te...Aim: To study the influences of dibutyryl cyclic adenosine monophosphate (dbcAMP) and forskolin on human sperm motility in vitro. Methods: Semen samples, aseptically obtained by masturbation and prepared by swim-up technique from 20 fertile men, were incubated with different concentrations of dbcAMP and forskolin at 37 癈. Measurements were carried out after 10 min, 20 min, 30 min and 60 min incubation. Motility parameters were estimated by using an automatic analyzing system. Results: Treatment with dbcAMP or forskolin resulted in a significant increase in sperm motility and progressive motility. The larger the concentrations of dbcAMP or forskolin, the greater the effect appeared. The straight linear velocity and curvilinear velocity were not affected by both agents. Conclusion: dbcAMP and forskolin increase the motility and progressive motility of human sperm in vitro.展开更多
For providing some experimental basis in establishing malignant phenotypic reversed indexes of gastric carcinoma cells, human gastric adenocar-cinoma cell line MGc80-3 was induced by dBcAMP in vitro to appraise the ef...For providing some experimental basis in establishing malignant phenotypic reversed indexes of gastric carcinoma cells, human gastric adenocar-cinoma cell line MGc80-3 was induced by dBcAMP in vitro to appraise the effect of gastric carcinoma cell differentiation by chemical inducers.Under light microscope, MGc80-3 cells, after treated with 1 mM dBcAMP, tended to be flat and disperse, and their volume gradually enlarged, with their uncleus relatively smaller and their shape rather regular. Morphological changes, like norma differentiated epithelial cells, were observed. The cells attached firmly, grew slowly, their growth curve showed inhibitory rate amounted to 52.87%, and cellular division exponent displayed their peak value 1.5 times less than that of MGc80-3 cells. It was clear that dBcAMP could effectively inhibit the multiplication activity of MGc80-3 cells. After dBcAMP treatment, remarkable changes of cell surface charges was indicated by cell electrophoresis, the ratio dropped to 3.043 from 3.988, and their re-tardant ratio reached up to 31.2%. cAMP content in cells after this treatment, detected by cAMP and cGMP radioimmunoassay, was enhanced by 2.42 times, and cAMP/cGMP ratio, by 1.73 times. Thus, cAMP level within MGc80-3 cells was raised obviously by dBcAMP. Heterotransplantation experiments showed that tuntorigenic rate of MGc80-5 cells (transplanted subcutaneously to BALB/c mice) amounted to 100%, and that of the cells after this treatment was only 5.6%. Their tumorigenic ability was extremely reduced.These results confirmed that dBcAMP was able to change malignant phenotypic characteristics of MGc80-3 cells and produce a reversed alteration: Thus, it has a remarkable inductive effect in differentiating gastric carcinoma cells. All these characteristics were also considered as the reference indexes in appraising reversed effect for the homologous cancer cells.展开更多
Dibutyryl adenosine cyclophosphate(dbc AMP-Ca), an analog of cyclic adenosine monophosphate(c AMP), plays greater roles in regulating physiological activities and energy metabolism than c AMP. The aim of this study wa...Dibutyryl adenosine cyclophosphate(dbc AMP-Ca), an analog of cyclic adenosine monophosphate(c AMP), plays greater roles in regulating physiological activities and energy metabolism than c AMP. The aim of this study was to investigate the effect of oral administration of dbc AMP-Ca on growth performance and fatty acids metabolism in weaning piglets. A total of 14 early weaning piglets(7 ± 1 d of age,3.31 ± 0.09 kg, Landrace ? Large White ? Duroc) were randomly divided into 2 groups: control group and dbc AMP-Ca group, and the piglets received 7 m L of 0.9% Na Cl or 1.5 mg dbc AMP-Ca dissolved in 7 m L of 0.9% Na Cl per day for 10 d, respectively. The results showed that the average daily gain(ADG)increased by 109.17%(P < 0.05) in the dbc AMP-Ca group compared with the control group. Besides,dbc AMP-Ca significantly decreased blood high density lipoprotein cholesterol(HDLC) concentration(P < 0.05) and significantly increased blood low density lipoprotein cholesterol(LDLC) concentration(P < 0.05) compared with the control group. Further, liver C18:2 n6 t content significantly increased in dbc AMP-Ca group(P < 0.05) compared with the control group. With the increase of C18:2 n6 t content,the m RNA expression levels of peroxisome proliferator-activated receptor a(PPARa) and hormone sensitive glycerol three lipase(HSL), of which genes are related to lipid metabolism, were also significantly increased(P < 0.05 or P < 0.01). All of the results indicated that dbc AMP-Ca improved the ADG, which was probably done by regulating fatty acids metabolism in the liver of weaning piglets.展开更多
基金Supported by Science and Technology Commission of Shanghai,No.13ZR1406700 and No.13DZ1930908
文摘AIM: To investigate whether the regulation of aquaporin 3 (AQP3) and AQP9 induced by Auphen and dibutyryl cAMP (dbcAMP) inhibits hepatic tumorigenesis.METHODS: Expression of AQP3 and AQP9 was detected by Western blot, immunohistochemistry (IHC), and RT-PCR in HCC samples and paired non-cancerous liver tissue samples from 30 hepatocellular carcinoma (HCC) patients. A xenograft tumor model was used in vivo. Nine nude mice were divided into control, Auphen-treated, and dbcAMP-treated groups (n = 3 for each group). AQP3 and AQP9 protein expression after induction of xenograft tumors was detected by IHC and mRNA by RT-PCR analysis. The terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay and histological evaluation were used to detect apoptosis of tumor cells, and the concentration of serum α-fetoprotein (AFP) was measured using RT-PCR and an ELISA kit.RESULTS: The volumes and weights of tumors decreased significantly in the Auphen- and dbcAMP-treated mice compared with the control mice (P < 0.01). The levels of AQP3 were significantly lower in the Auphen treatment group, and levels of AQP9 were significantly higher in thedbcAMP treatment mice than in the control mice (P < 0.01). The reduction of AQP3 by Auphen and increase of AQP9 by dbcAMP in nude mice suppressed tumor growth of HCC, which resulted in reduced AFP levels in serum and tissues, and apoptosis of tumor cells in the Auphen- and dbcAMP-treated mice, when compared with control mice (P < 0.01). Compared with para-carcinoma tissues, AQP3 expression increased in tumor tissues whereas the expression of AQP9 decreased. By correlating clinicopathological and expression levels, we demonstrated that the expression of AQP3 and AQP9 was correlated with clinical progression of HCC and disease outcomes.CONCLUSION: AQP3 increases in HCC while AQP9 decreases. Regulation of AQP3 and AQP9 expression by Auphen and dbcAMP inhibits the development and growth of HCC.
文摘Aim: To study the influences of dibutyryl cyclic adenosine monophosphate (dbcAMP) and forskolin on human sperm motility in vitro. Methods: Semen samples, aseptically obtained by masturbation and prepared by swim-up technique from 20 fertile men, were incubated with different concentrations of dbcAMP and forskolin at 37 癈. Measurements were carried out after 10 min, 20 min, 30 min and 60 min incubation. Motility parameters were estimated by using an automatic analyzing system. Results: Treatment with dbcAMP or forskolin resulted in a significant increase in sperm motility and progressive motility. The larger the concentrations of dbcAMP or forskolin, the greater the effect appeared. The straight linear velocity and curvilinear velocity were not affected by both agents. Conclusion: dbcAMP and forskolin increase the motility and progressive motility of human sperm in vitro.
文摘For providing some experimental basis in establishing malignant phenotypic reversed indexes of gastric carcinoma cells, human gastric adenocar-cinoma cell line MGc80-3 was induced by dBcAMP in vitro to appraise the effect of gastric carcinoma cell differentiation by chemical inducers.Under light microscope, MGc80-3 cells, after treated with 1 mM dBcAMP, tended to be flat and disperse, and their volume gradually enlarged, with their uncleus relatively smaller and their shape rather regular. Morphological changes, like norma differentiated epithelial cells, were observed. The cells attached firmly, grew slowly, their growth curve showed inhibitory rate amounted to 52.87%, and cellular division exponent displayed their peak value 1.5 times less than that of MGc80-3 cells. It was clear that dBcAMP could effectively inhibit the multiplication activity of MGc80-3 cells. After dBcAMP treatment, remarkable changes of cell surface charges was indicated by cell electrophoresis, the ratio dropped to 3.043 from 3.988, and their re-tardant ratio reached up to 31.2%. cAMP content in cells after this treatment, detected by cAMP and cGMP radioimmunoassay, was enhanced by 2.42 times, and cAMP/cGMP ratio, by 1.73 times. Thus, cAMP level within MGc80-3 cells was raised obviously by dBcAMP. Heterotransplantation experiments showed that tuntorigenic rate of MGc80-5 cells (transplanted subcutaneously to BALB/c mice) amounted to 100%, and that of the cells after this treatment was only 5.6%. Their tumorigenic ability was extremely reduced.These results confirmed that dBcAMP was able to change malignant phenotypic characteristics of MGc80-3 cells and produce a reversed alteration: Thus, it has a remarkable inductive effect in differentiating gastric carcinoma cells. All these characteristics were also considered as the reference indexes in appraising reversed effect for the homologous cancer cells.
基金jointly supported by National Key Research and Development Program of China (2016YFD0501209, and 2016YFD0500504)the Earmarked Fund for China Agriculture Research System (CARS-35)+1 种基金the Major Project of Hunan Province (2016NK2124 2015NK1002)
文摘Dibutyryl adenosine cyclophosphate(dbc AMP-Ca), an analog of cyclic adenosine monophosphate(c AMP), plays greater roles in regulating physiological activities and energy metabolism than c AMP. The aim of this study was to investigate the effect of oral administration of dbc AMP-Ca on growth performance and fatty acids metabolism in weaning piglets. A total of 14 early weaning piglets(7 ± 1 d of age,3.31 ± 0.09 kg, Landrace ? Large White ? Duroc) were randomly divided into 2 groups: control group and dbc AMP-Ca group, and the piglets received 7 m L of 0.9% Na Cl or 1.5 mg dbc AMP-Ca dissolved in 7 m L of 0.9% Na Cl per day for 10 d, respectively. The results showed that the average daily gain(ADG)increased by 109.17%(P < 0.05) in the dbc AMP-Ca group compared with the control group. Besides,dbc AMP-Ca significantly decreased blood high density lipoprotein cholesterol(HDLC) concentration(P < 0.05) and significantly increased blood low density lipoprotein cholesterol(LDLC) concentration(P < 0.05) compared with the control group. Further, liver C18:2 n6 t content significantly increased in dbc AMP-Ca group(P < 0.05) compared with the control group. With the increase of C18:2 n6 t content,the m RNA expression levels of peroxisome proliferator-activated receptor a(PPARa) and hormone sensitive glycerol three lipase(HSL), of which genes are related to lipid metabolism, were also significantly increased(P < 0.05 or P < 0.01). All of the results indicated that dbc AMP-Ca improved the ADG, which was probably done by regulating fatty acids metabolism in the liver of weaning piglets.
文摘目的:建立具有分化成γ-氨基丁酸能神经元潜能的神经干细胞模型,为γ-氨基丁酸能神经元退行性疾病的研究提供适宜的研究载体。方法:应用双丁酰环腺苷酸(dbcAMP)诱导人神经母细胞瘤细胞(SH-SY5Y细胞),分为0mmol·L-1 dbcAMP组(对照组)和0.3、0.6、1.0及2.0dbcAMP组,观察诱导后各组SH-SY5Y细胞的形态变化;采用Imge-Pro Plus 5.0软件测量神经元样细胞神经突起长度,计算神经突起>30μm细胞所占细胞总数的百分率;采用免疫荧光细胞化学技术检测γ-氨基丁酸能神经元标志性蛋白——谷氨酸脱羧酶65(GAD65)的表达,并计算其免疫反应阳性率。结果:形态学观察,对照组SH-SY5Y细胞呈多边形、圆形或梭型,胞膜光滑,边界清晰;各浓度dbcAMP组随着dbcAMP浓度的增加和诱导时间的延长,SH-SY5Y细胞胞体变小、突起变长;1.0mmol·L-1 dbcAMP组细胞互相交织,表现出成熟神经元的表型。SH-SY5Y细胞诱导培养72h后,与对照组(31.4%±4.2%)比较,0.3、0.6、1.0和2.0dbcAMP组神经突起>30μm细胞所占细胞总数的百分率(40.1%±5.7%、47.5%±6.2%、73.1%±3.2%和74.3%±6.1%)明显升高(P<0.05或P<0.01)。SH-SY5Y细胞诱导培养72h后,与对照组(10.2%±2.1%)比较,0.3、0.6、1.0和2.0dbcAMP组GAD65阳性细胞表达率(22.1%±2.4%、46.9%±3.2%、70.7%±3.4%和72.3%±3.7%)明显升高(P<0.05或P<0.01)。结论:SH-SY5Y细胞具有分化为γ-氨基丁酸能神经元样细胞的潜能,1.0mmol·L-1是dbcAMP的最佳诱导浓度。
