Aim:To elucidate the distribution and regulation of Neuropeptide Y(NPY)gene expression in testes under physiologi- cal and pathophysiological conditions,such as testicular development,fasting and experimental cryptorc...Aim:To elucidate the distribution and regulation of Neuropeptide Y(NPY)gene expression in testes under physiologi- cal and pathophysiological conditions,such as testicular development,fasting and experimental cryptorchidism. Methods:In the first experiment,C57BL/6J male mice at the ages of 3 days as well as 2,3,5 and 8 weeks were used. In the second and third experiments,adult C57BL/6J male mice were subjected to fasting for 48 h and experimental cryptorchidism for 72 h.The NPY transcripts were detected by isotopic in situ hybridization histochemistry.Results: The NPY transcripts were exclusively expressed in the interstitial cells regardless of the age groups and experimental conditions.The NPY mRNA levels were found to increase significantly with age(from the neonatal to prepubertal period [P<0.01] and from the prepubetal to postpubertal period [P<0.01]).Food deprivation for 48 h resulted in a significant increase in the NPY mRNA levels in the arcuate nucleus of the hypothalamus(P<0.01),but not in the testes.Experimental cryptorchidism for 72 h failed to regulate the NPY gene expression in the testes.Conclusion: NPY gene expression is distributed in Leydig cells and increases in line with testicular development.The regulation of testicular NPY is different from that of hypothalamic NPY.(Asian J Androl 2006 Jul;8:443-449)展开更多
Objective: A traditional Chinese medicine Tian Gui Recipe (TGR) has been used to effectively treat clomiphene resistant anovulatory disease and obesity, especially in polycystic ovary syndrome (PCOS) cases with hyperi...Objective: A traditional Chinese medicine Tian Gui Recipe (TGR) has been used to effectively treat clomiphene resistant anovulatory disease and obesity, especially in polycystic ovary syndrome (PCOS) cases with hyperinsulinemia. The effect of TGR on obesity and anovulation was investigated in androgen-sterilized rats (ASR). Methods: Female SD rats at the age of 9 days were divided into 3 groups. Group ASR (n=15): anovulation was demonstrated at the age of 70 days by vaginal smear in rats while 1. 25 mg of testosterone propionate was injected subcutaneously on its 9th neonatal day. Group A+H (n = 25): TGR were administered to ASR at the age of 80 days for 3 weeks. Rats with regular estrous cycle and ovulation after herbal treatment were included in this group. Group C (n = 15): normal ovulated rats were recruited. Around the age of 112 days or on proestrous day, all rats were sacrificed under anesthesia. Serum leptin, testosterone (T), estrogen (E2), follicular stimulating hormone (FSH), luteinizing hormone (LH) levels were measured with radioimmunoassay (RIA). Double immunofluorescent staining combined with confocal laser scanning microscope and dual in situ hybridization were carried out on sections through areas of arcuate nucleus (ARC) to determine whether estrogen receptor (ER) immunoreactivity (IR) or long form of leptin receptor (OB-Rb) mRNA were expressed in neuropeptide Y (NPY) immuno-and mRNA-containing neurons, and its relationship to proopiomelanocortin (POMC) mRNA-containing neurons. Immunohistochemistry and in situ hybridization were used to observe the levels of ER, NPY, gonadotrophin releasing hormone (GnRH) and gene expression levels on NPY, OB-Rb, POMC. Meanwhile, the criteria of energy state, including daily food intake, retroperitoneal fat depot pad and body weight, were measured and evaluated. Values of immunohistochemistry and in situ hybridization were expressed in mean optic density (MOD). Results: Seventeen out of the 25 rats (68%) in Group A+H were ovulated after herbal treatment. ASR characterized with significantly high metabolic rate, energy imbalance and obesity (P<0. 01, P<0. 01, P<0. 01). The mean serum leptin, T, E2 levels of group ASR were significantly higher than those of Group C and Group A+H (P< 0. 01,P<0. 01 ), while FSH, LH levels were lower (P<0. 01, P<0. 01 ). ERs were proved to be distributing in NPY-containing neurons, OB-Rb coexpressed in NPY-syn- thesizing neurons, and both NPY and POMC mRNA expressing neurons overlapped in ARC. Compared to Group C, levels of NPY mRNA and NPY, POMC mRNA expressions were increased (P<0. 01, P<0. 01, P<0. 01 ), OB-Rb mRNA, ER and GnRH immunoreactive expression were decreased significantly in the hypothalamus of ASR (P<0. 01, P<0. 01, P<0. 01 ). These dramatic neuroendocrine changes became normal in ovulated ASR after the herbal treatment (all P>0. 05). Conclusion: The elevated peripheral E2 caused by high T and leptin levels in ASR may down-regulate their corresponding receptors oriented on hypothalamic NPY- containing neurons respectively, therefore challenge the NPY mRNA and NPY, POMC mRNA overexpressions. The overexpression, of NPY, POMC gene transcription and translation stimulate food intake, promote the deposit of adipocyte tissue, and inhibit GnRH expression and GnRH/FSH, LH release, which contribute to the occurrence of obesity and anovulation in ASR. TGR may reverse these abnormal neuroendocrine cascades by lowering the levels of T, E2 and leptin.展开更多
Objectives: A traditional Chinese medicine “Tian Gui Recipe (TGR)” has been used to effectively treat clomiphene resistant anovulatory disease and obesity, especially in polycystic ovary syndrome (PCOS) cases with h...Objectives: A traditional Chinese medicine “Tian Gui Recipe (TGR)” has been used to effectively treat clomiphene resistant anovulatory disease and obesity, especially in polycystic ovary syndrome (PCOS) cases with hyperinsulinemia. The effect of TGR on obesity and anovulation was investigated in androgen-sterilized rats (ASR).Methods: Female SD rats at the age of 9 days were divided into 3 groups. Group ASR (n=15): anovulation was demonstrated at the age of 70 days by vaginal smear in rats while 1.25 mg of testosterone propionate was injected subcutaneously on its 9th neonatal day. Group A+H (n=25): TGR were administered to ASR at the age of 80 days for 3 weeks. Rats with regular estrous cycle and ovulation after herbal treatment were included in this group. Group C (n=15): normal ovulated rats were recruited. Around the age of 112 days or on proestrous day, all rats were sacrificed under anesthesia. Serum leptin, testosterone (T), estrogen (E2), follicular stimulating hormone (FSH), luteinizing hormone (LH) levels were measured with radioimmunoassay (RIA). Double immunofluorescent staining combined with confocal laser scanning microscope and dual in situ hybridization were carried out on sections through areas of arcuate nucleus (ARC) to determine whether estrogen receptor (ER) immunoreactivity (IR) or long form of leptin receptor (OB-Rb) mRNA were expressed in neuropeptide Y (NPY) immuno- and mRNA-containing neurons, and their relationship to proopiomelanocortin (POMC) mRNA-containing neurons. Immunohistochemistry and in situ hybridization were used to observe the levels of ER-, NPY-,gonadotrophin releasing hormone (GnRH)-IR and gene expression levels of NPY, OB-Rb, and POMC in ARC. Meanwhile, the criteria of energy state,including daily food intake, retroperitoneal fat depot pad and body weight,were measured and evaluated. Values of immunohistochemistry and in situ hybridization were expressed in mean optic density (MOD).Results:Seventeen out of the 25 rats (68%) in Group A+H were ovulated after herbal treatment. ASR characterized with significantly high metabolic rate, energy imbalance and obesity (P<0.01,P<0.01,P<0.01).The mean serum leptin, T, E2 levels of ASR were significantly higher than that of Group C and Group A+H (P<0.01,P<0.01), while being contraryto that of FSH, LH levels (P<0.01,P<0.01). ERs were proved distributing in NPY-containing neurons, OB-Rb coexpressed in NPY-synthesizing neurons, and both NPY- and POMC- mRNA expressing neurons overlapped in ARC. Compared to Group C, levels of NPY mRNA and NPY, POMC mRNA expressions increased (P<0.01,P<0.01,P<0.01),OB-Rb mRNA, ER and GnRH immunoreactive expression decreased significantly in the hypothalamus of ASR (P<0.01,P<0.01,P<0.01).These dramatic neuroendocrine changes became normal in ovulated ASR after the herbal treatment (all P>0.05).Conclusion: The elevated peripheral estrogen caused by high testosterone and leptin levels in ASR may down-regulate their corresponding receptors oriented on hypothalamic NPY-containing neurons respectively, therefore challenge the NPY mRNA and NPY,POMC mRNA overexpression. The overexpression of NPY, POMC gene transcription and translation stimulate food intake, promote the deposit of adipocyte tissue, and inhibit GnRH expression and GnRH/FSH, LH release, which contribute to the occurrence of obesity and anovulation in ASR. “TGR” may reverse these abnormal neuroendocrine cascades by lowering levels of testosterone, estrogen and leptin.展开更多
In recent years, the endothelial cell has become the popular subject of many scientificresearch fields such as cardiovascular and cancer pathology. The results reported up to nowindicate that the endothelial cell has ...In recent years, the endothelial cell has become the popular subject of many scientificresearch fields such as cardiovascular and cancer pathology. The results reported up to nowindicate that the endothelial cell has the complex funtion in synthesis and metabolism, andtherefore plays an important role in the regulation of vascular function. Burnstock’s labfirst observed that many kinds of vasoactive substances are localized in the endothelial展开更多
OBJECTIVE: To study the relationship between neuropeptide Y (NPY) and diabetes by examining the content and distribution of NPY and its mRNA expression in the hypothalamus and pancreas of STZ-diabetic rats. METHODS: T...OBJECTIVE: To study the relationship between neuropeptide Y (NPY) and diabetes by examining the content and distribution of NPY and its mRNA expression in the hypothalamus and pancreas of STZ-diabetic rats. METHODS: Thirty Wistar rats were randomly divided into 3 groups (diabetic group, diabetic insulin treatment group, and control group). After feeding for 24 weeks, the rats were sacrificed. The expression of NPY in the hypothalamus and pancreas was detected with immunohistochemistry and in situ hybridization. RESULTS: (1) The hypothalamic content of NPY and its mRNA were significantly increased in STZ-diabetic rats in comparison with normal controls. Increased expression of NPY mRNA was found only in the arcuate nucleus and not in the paraventricular nucleus in diabetic rats, suggesting that NPY was produced in the arcuate nucleus. (2) The hypothalamic content of NPY and its mRNA in STZ-diabetic rats were visibly reduced after insulin treatment compared with that in untreated diabetic rats. This supports the hypothesis that insulin deficiency in the brain may be responsible for increased hypothalamic NPY gene expression in diabetic rats. (3) The increase of hypothalamic NPY in STZ diabetic rats associated with hyperphagia and polydipsia could be reversed by insulin replacement, suggesting that increased hypothalamic NPY contributes to the pathophysiological progress of the diabetic state. (4) The present study demonstrated for the first time that the content of NPY and its mRNA in the pancreas was increased in STZ-diabetic rats, and that the distribution of NPY-positive cell in islets was changed from the periphery to the whole islet. The content and distribution of NPY and its mRNA in islets were not changed by insulin treatment. CONCLUSION: Increased NPY in the hypothalamus results in hypophagia and polydipsia, while the implication of increased NPY in the pancreas of diabetic rats is not clear.展开更多
文摘Aim:To elucidate the distribution and regulation of Neuropeptide Y(NPY)gene expression in testes under physiologi- cal and pathophysiological conditions,such as testicular development,fasting and experimental cryptorchidism. Methods:In the first experiment,C57BL/6J male mice at the ages of 3 days as well as 2,3,5 and 8 weeks were used. In the second and third experiments,adult C57BL/6J male mice were subjected to fasting for 48 h and experimental cryptorchidism for 72 h.The NPY transcripts were detected by isotopic in situ hybridization histochemistry.Results: The NPY transcripts were exclusively expressed in the interstitial cells regardless of the age groups and experimental conditions.The NPY mRNA levels were found to increase significantly with age(from the neonatal to prepubertal period [P<0.01] and from the prepubetal to postpubertal period [P<0.01]).Food deprivation for 48 h resulted in a significant increase in the NPY mRNA levels in the arcuate nucleus of the hypothalamus(P<0.01),but not in the testes.Experimental cryptorchidism for 72 h failed to regulate the NPY gene expression in the testes.Conclusion: NPY gene expression is distributed in Leydig cells and increases in line with testicular development.The regulation of testicular NPY is different from that of hypothalamic NPY.(Asian J Androl 2006 Jul;8:443-449)
文摘Objective: A traditional Chinese medicine Tian Gui Recipe (TGR) has been used to effectively treat clomiphene resistant anovulatory disease and obesity, especially in polycystic ovary syndrome (PCOS) cases with hyperinsulinemia. The effect of TGR on obesity and anovulation was investigated in androgen-sterilized rats (ASR). Methods: Female SD rats at the age of 9 days were divided into 3 groups. Group ASR (n=15): anovulation was demonstrated at the age of 70 days by vaginal smear in rats while 1. 25 mg of testosterone propionate was injected subcutaneously on its 9th neonatal day. Group A+H (n = 25): TGR were administered to ASR at the age of 80 days for 3 weeks. Rats with regular estrous cycle and ovulation after herbal treatment were included in this group. Group C (n = 15): normal ovulated rats were recruited. Around the age of 112 days or on proestrous day, all rats were sacrificed under anesthesia. Serum leptin, testosterone (T), estrogen (E2), follicular stimulating hormone (FSH), luteinizing hormone (LH) levels were measured with radioimmunoassay (RIA). Double immunofluorescent staining combined with confocal laser scanning microscope and dual in situ hybridization were carried out on sections through areas of arcuate nucleus (ARC) to determine whether estrogen receptor (ER) immunoreactivity (IR) or long form of leptin receptor (OB-Rb) mRNA were expressed in neuropeptide Y (NPY) immuno-and mRNA-containing neurons, and its relationship to proopiomelanocortin (POMC) mRNA-containing neurons. Immunohistochemistry and in situ hybridization were used to observe the levels of ER, NPY, gonadotrophin releasing hormone (GnRH) and gene expression levels on NPY, OB-Rb, POMC. Meanwhile, the criteria of energy state, including daily food intake, retroperitoneal fat depot pad and body weight, were measured and evaluated. Values of immunohistochemistry and in situ hybridization were expressed in mean optic density (MOD). Results: Seventeen out of the 25 rats (68%) in Group A+H were ovulated after herbal treatment. ASR characterized with significantly high metabolic rate, energy imbalance and obesity (P<0. 01, P<0. 01, P<0. 01). The mean serum leptin, T, E2 levels of group ASR were significantly higher than those of Group C and Group A+H (P< 0. 01,P<0. 01 ), while FSH, LH levels were lower (P<0. 01, P<0. 01 ). ERs were proved to be distributing in NPY-containing neurons, OB-Rb coexpressed in NPY-syn- thesizing neurons, and both NPY and POMC mRNA expressing neurons overlapped in ARC. Compared to Group C, levels of NPY mRNA and NPY, POMC mRNA expressions were increased (P<0. 01, P<0. 01, P<0. 01 ), OB-Rb mRNA, ER and GnRH immunoreactive expression were decreased significantly in the hypothalamus of ASR (P<0. 01, P<0. 01, P<0. 01 ). These dramatic neuroendocrine changes became normal in ovulated ASR after the herbal treatment (all P>0. 05). Conclusion: The elevated peripheral E2 caused by high T and leptin levels in ASR may down-regulate their corresponding receptors oriented on hypothalamic NPY- containing neurons respectively, therefore challenge the NPY mRNA and NPY, POMC mRNA overexpressions. The overexpression, of NPY, POMC gene transcription and translation stimulate food intake, promote the deposit of adipocyte tissue, and inhibit GnRH expression and GnRH/FSH, LH release, which contribute to the occurrence of obesity and anovulation in ASR. TGR may reverse these abnormal neuroendocrine cascades by lowering the levels of T, E2 and leptin.
基金a grant from the Planning of the Leading Medical Discipline-Gynecology in Integrated Traditional Chinese and Western Medici*ne(No.96001),Shanghai Health Bureau.
文摘Objectives: A traditional Chinese medicine “Tian Gui Recipe (TGR)” has been used to effectively treat clomiphene resistant anovulatory disease and obesity, especially in polycystic ovary syndrome (PCOS) cases with hyperinsulinemia. The effect of TGR on obesity and anovulation was investigated in androgen-sterilized rats (ASR).Methods: Female SD rats at the age of 9 days were divided into 3 groups. Group ASR (n=15): anovulation was demonstrated at the age of 70 days by vaginal smear in rats while 1.25 mg of testosterone propionate was injected subcutaneously on its 9th neonatal day. Group A+H (n=25): TGR were administered to ASR at the age of 80 days for 3 weeks. Rats with regular estrous cycle and ovulation after herbal treatment were included in this group. Group C (n=15): normal ovulated rats were recruited. Around the age of 112 days or on proestrous day, all rats were sacrificed under anesthesia. Serum leptin, testosterone (T), estrogen (E2), follicular stimulating hormone (FSH), luteinizing hormone (LH) levels were measured with radioimmunoassay (RIA). Double immunofluorescent staining combined with confocal laser scanning microscope and dual in situ hybridization were carried out on sections through areas of arcuate nucleus (ARC) to determine whether estrogen receptor (ER) immunoreactivity (IR) or long form of leptin receptor (OB-Rb) mRNA were expressed in neuropeptide Y (NPY) immuno- and mRNA-containing neurons, and their relationship to proopiomelanocortin (POMC) mRNA-containing neurons. Immunohistochemistry and in situ hybridization were used to observe the levels of ER-, NPY-,gonadotrophin releasing hormone (GnRH)-IR and gene expression levels of NPY, OB-Rb, and POMC in ARC. Meanwhile, the criteria of energy state,including daily food intake, retroperitoneal fat depot pad and body weight,were measured and evaluated. Values of immunohistochemistry and in situ hybridization were expressed in mean optic density (MOD).Results:Seventeen out of the 25 rats (68%) in Group A+H were ovulated after herbal treatment. ASR characterized with significantly high metabolic rate, energy imbalance and obesity (P<0.01,P<0.01,P<0.01).The mean serum leptin, T, E2 levels of ASR were significantly higher than that of Group C and Group A+H (P<0.01,P<0.01), while being contraryto that of FSH, LH levels (P<0.01,P<0.01). ERs were proved distributing in NPY-containing neurons, OB-Rb coexpressed in NPY-synthesizing neurons, and both NPY- and POMC- mRNA expressing neurons overlapped in ARC. Compared to Group C, levels of NPY mRNA and NPY, POMC mRNA expressions increased (P<0.01,P<0.01,P<0.01),OB-Rb mRNA, ER and GnRH immunoreactive expression decreased significantly in the hypothalamus of ASR (P<0.01,P<0.01,P<0.01).These dramatic neuroendocrine changes became normal in ovulated ASR after the herbal treatment (all P>0.05).Conclusion: The elevated peripheral estrogen caused by high testosterone and leptin levels in ASR may down-regulate their corresponding receptors oriented on hypothalamic NPY-containing neurons respectively, therefore challenge the NPY mRNA and NPY,POMC mRNA overexpression. The overexpression of NPY, POMC gene transcription and translation stimulate food intake, promote the deposit of adipocyte tissue, and inhibit GnRH expression and GnRH/FSH, LH release, which contribute to the occurrence of obesity and anovulation in ASR. “TGR” may reverse these abnormal neuroendocrine cascades by lowering levels of testosterone, estrogen and leptin.
文摘In recent years, the endothelial cell has become the popular subject of many scientificresearch fields such as cardiovascular and cancer pathology. The results reported up to nowindicate that the endothelial cell has the complex funtion in synthesis and metabolism, andtherefore plays an important role in the regulation of vascular function. Burnstock’s labfirst observed that many kinds of vasoactive substances are localized in the endothelial
基金ThisworkwassupportedbyagrantofNationalNatureScienceFoundationofChina (No 3 9770 3 5 1) agrantofDoctoralTrainingFoundationoftheMinistryofHealth ,hina (No .19980 3 0 3 4)
文摘OBJECTIVE: To study the relationship between neuropeptide Y (NPY) and diabetes by examining the content and distribution of NPY and its mRNA expression in the hypothalamus and pancreas of STZ-diabetic rats. METHODS: Thirty Wistar rats were randomly divided into 3 groups (diabetic group, diabetic insulin treatment group, and control group). After feeding for 24 weeks, the rats were sacrificed. The expression of NPY in the hypothalamus and pancreas was detected with immunohistochemistry and in situ hybridization. RESULTS: (1) The hypothalamic content of NPY and its mRNA were significantly increased in STZ-diabetic rats in comparison with normal controls. Increased expression of NPY mRNA was found only in the arcuate nucleus and not in the paraventricular nucleus in diabetic rats, suggesting that NPY was produced in the arcuate nucleus. (2) The hypothalamic content of NPY and its mRNA in STZ-diabetic rats were visibly reduced after insulin treatment compared with that in untreated diabetic rats. This supports the hypothesis that insulin deficiency in the brain may be responsible for increased hypothalamic NPY gene expression in diabetic rats. (3) The increase of hypothalamic NPY in STZ diabetic rats associated with hyperphagia and polydipsia could be reversed by insulin replacement, suggesting that increased hypothalamic NPY contributes to the pathophysiological progress of the diabetic state. (4) The present study demonstrated for the first time that the content of NPY and its mRNA in the pancreas was increased in STZ-diabetic rats, and that the distribution of NPY-positive cell in islets was changed from the periphery to the whole islet. The content and distribution of NPY and its mRNA in islets were not changed by insulin treatment. CONCLUSION: Increased NPY in the hypothalamus results in hypophagia and polydipsia, while the implication of increased NPY in the pancreas of diabetic rats is not clear.
