促肾上腺皮质激素释放因子（CRF）对人胶质瘤细胞株U87CRFR1表达的比较研究
|
摘要
促肾上腺皮质激素释放因子（Corticotropin-releasing factor，CRF）是一种重要的神经内分泌肽，主要生理作用是刺激腺垂体合成和分泌促肾上腺皮质激素（Adrenocorticotropic hormon，ACTH）及其有关多肽。CRF及其肽家族共同调节机体在应激状态下丘脑—垂体—肾上腺（Hypothalamus-pituitary-adpenal axis，HPA）轴的功能,并且协调应激状态下内分泌系统、自主神经系统、免疫系统、生理学及行为学等功能，并对心血管、胃肠道、生殖、皮肤等有调节作用。CRF及其相关肽通过其受体(Corticotropin releasing factor receptor，CRFR)发挥作用。CRF受体属于G-蛋白偶联受体,通过腺苷酸环化酶的活化发挥作用。促肾上腺皮质激素释放激素受体1（Corticotropin releasing factor receptor1，CRFR1)是HPA轴中最主要的受体，主要介导经典的CRF促进垂体ACTH释放的作用，参与应激反应。而在人类中枢系统的一些恶性肿瘤组织中亦发现存在有CRFR1受体的过度表达。但在CRF及其受体与胶质瘤的研究中还未曾发现它们之间存在有某些确定的理论关系。
目的：探讨人胶质瘤细胞株U87中CRFR1受体表达及其应用不同浓度CRF作用U87细胞后CRFR1受体的表达情况。
方法：
1从培养的U87细胞中提取总RNA（ribonucleic acid），设计CRFR1引物，进行RT-PCR扩增，凝胶电泳分析结果。
2不同浓度CRF作用于胶质瘤细胞U87作用。将U87传代后随机分为四组，每组分别加入不同浓度的CRF（10-7,10-9,10-12mol/L）和不加CRF（对照组）的DMEM（含10%牛清+青链霉素）培养基，在有盖玻片的六孔培养板中继续培养U87细胞。培养12h后，每个浓度分别各取出两个孔中的细胞，进行细胞的免疫化学染色。剩余孔中细胞继续培养，在24h后以同样的方法再取出一批进行细胞免疫组化。最后一批于48h后取出行细胞免疫组化。实验重复三次。最后以细胞免疫组化结果中CRFR1表达的阳性率为结果进行统计学分析。
结果：
1RT-PCR的结果显示人胶质瘤U87中存在CRFR1mRNA表达。2.进行免疫细胞化学染色，结果显示所有组中均表达CRFR1阳性，阳性反应物位于胞浆，在细胞核周边呈现清晰淡黄色或棕黄色。以CRFR1表达的阳性率为统计数据：
2.1采用单因素方差分析不同CRF浓度对CRFR1表达的影响,每个实验组和对照组相比较，经统计学处理结果如下：12h（F=8.099，P<0.05）；24h（F=51.665，P<0.05）；48h（F=14.531，P<0.05）。可以认为不同CRF浓度对U87作用对CRFR1阳性表达率存在影响。在作用12或48h，CRF10-9mol/L和其它各浓度组比较均有差别，而其它各浓度组间比较均无差别。在作用24小时，对照组和CRF10-9mol/L比较有差别；对照组和CRF10-7mol/L，CRF10-12mol/L比较无差别; CRF10-7mol/L和CRF10-12mol/L组比较有差别。
2.2采用单因素方差分析分析不同作用时间对CRFR1表达阳性率的影响，经统计学处理结果如下：CRF10-7mol/L作用不同的时间对CRFR1表达阳性率无影响（F=1.839，P=0.238）；CRF10-12mol/L作用不同的时间对CRFR1表达阳性率无影响(F=3.884,P=0.083）；CRF10-9mol/L作用不同的时间对CRFR1表达阳性率有影响（F=90.66,P=0.0001），不同时间的多重比较中任意两组比较均有差别。
2.3对不同浓度CRF及其在不同的时间点相互交叉作用U87后CRFR1表达的阳性率数据进行析因分析；CRF不同的浓度对其CRFR1表达率有影响（F=21.185,P=0.0001）;不同的作用时间点对CRFR1表达率也有影响（F=27.954，P=0.0001）;并且不同的CRF浓度和不同的作用时间存在有交互作用（F=27.458，P=0.0001）；不同CRF药物浓度及不同的时间，CRFR1表达阳性细胞百分率有所差异，在31%～93%之间。并且表达强度亦有差别。在CRF10-9mol/L且24小时，结果显示为CRFR1表达阳性最强，呈棕褐色，并且表达阳性率最高，平均在87%左右。在CRF10-9mol/L作用48小时，CRFR1表达阳性率也较高，平均在68%左右。
结论：在RT-PCR结果中人胶质瘤U87中表达CRFR1mRNA。 CRF作用于胶质瘤U87细胞后，其CRFR1受体表达的阳性细胞率在统计学上有意义。CRF的浓度及其作用时间均可影响其CRFR1受体细胞阳性率的表达，并且不同浓度和时间存在交互作用。在CRF为10-9mol/L作用24小时时，CRFR1阳性细胞表达率最高，染色更明显。在CRF10-9mol/L时作用U87细胞，在24和48小时间上调了CRFR1受体的表达。但在其它浓度和时间点是这种现象则不明显，在CRF10-9mol/L时12小时表现为对CRFR1受体的下调。据我们的研究结果，推测CRF通过其受体CRFR1的表达对体外胶质瘤细胞的生长确实起到了一定影响的作用。
Corticotropin releasing factor（CRF）is an important neuroendocrinepeptide, and the main physiological function is to stimulate the pituitary glandsynthesis and secretion of adrenocorticotropic hormon（ACTH）and relatedpeptide. CRF and its peptide family common regulate the function ofhypothalamus-pituitary-adrenal (HPA) axis and coordinate the endocrinesystem, the autonomic nervous system, immune system, physiology andbehavior function when the body under stress. And, it has a moderating effecton cardiovascular, gastrointestinal, reproductive, skin, etc. CRF and its relatedpeptide play a role through its receptor（Corticotropin releasing factorreceptor，CRFR）. CRF receptors play a role through activation of adenylatecyclase, which belong to the G-protein coupling receptors. As the mainreceptor in the HPA axis, corticotropin releasing hormone receptor1(CRFR1)can promote the release of the pituitary ACTH to mediated CRF by the classicway under stress response. Overexpression of CRFR1recepor has also beenfound in so many human malignant tumors in the central nervous system.However, previous studies have not investigated the relationship of CRF andgliomas.
Objective：To explored the expression of CRFR1and the effcet of CRFinducing CRFR1in glioma cell line U87.
Methods：
1The total RNA were extracted from cultured U87cells. Primer of CRFR1were designed for the RT-PCR amplification. The product was analysis by gelelectrophoresis.
2Glioma cell lines U87were treated by different concentrations of CRF. U87cells was divided into four groups, which be dealed with by differentconcentrations of CRF (10-7,10-9,10-12mol/L) and control groups. Then U87 cells were cultured continuely in filled with coverslip of tin plates. Each twoholes were taken out to further immunostainned after12hours. The cells ofsurplus holes continue to culture. Another batch of cells were treated in thesame way after24hours. The last ones were treated in the same way after48hours. The experiment was repeated for three times. Finally, expression ofCRFR1positive rate were carried out by statistical analysis.
Results：
1CRFR1mRNA was expressed in human glioma cells U87by RT-PCR.2.CRFR1positive expression can be shown in every group byimmunocytochemistry. Positive protein located in the cytoplasm, andpresented a clear yellow or brown stain in the periphery of the nucleus.CRFR1-positive expression rates were carried out by statistically analysis.
2.1The single factor analysis were used for different concentration effect onthe expression the CRFR1. Compared with the control groups，differentconcentrations showed significant difference, which including12hours(F=8.099, P<0.05);24hours（F=51.665，P<0.05）;48hours (F=14.531,P<0.05). Compare the CRF10-9mol/L group with other concentration ones,there were difference at24hours and48hours. While compare with otherconcentration groups, there is no difference. At24hours, the control groupand CRF10-9mol/L group is significant difference. However, there is nodifference among the control group，CRF10-7mol/L group and CRF10-12mol/Lones. There is significant difference between the CRF10-7mol/L groups andCRF10-12mol/L ones.
2.2The single factor analysis were used for different time effect on theexpression the CRFR1. The statistics results are as follows：there is noinfluence to CRFR1expression positive rate in different times byCRF10-7mol/L treated（F=1.839，P=0.238）;and there is no influence to CRFR1expression positive rate in different times by CRF10-12mol/L treated（F=3.884,P=0.083）；there have influence to CRFR1expression positive ratein different times byCRF10-9mol/Ltreated(F=90.66,P=0.0001）, multiplecomparisons of the different time points, any two groups of comparisons are differences.
2.3Factorial analysis were carried out for different concentrations of CRFand different time to inducing CRFR1positive-expression rate. CRFR1expression rate were inducing by different concentration of CRF（F=21.185,P=0.0001）. CRFR1expression rate were influenced by different time（F=27.954, P=0.0001）; And, there are interaction between differentconcentration of CRF and different time（F=27.458, P=0.0001）. CRFR1expression rate were influenced by different concentration and time of CRF,which about31percent to93percent. And expression intensity was alsodifferences in conclusion. In CRF10-9mol/L and24hours, and the resultsshowed that CRFR1expression was strongest (the brown color)， andexpression of the highest positive rate,87%on average. In CRF10-9mol/L and48hours，expression of CRFR1-positive rate were68%on average.
Conclusion：The expression of CRFR1mRNA was found by RT-PCR inhuman glioma cells U87. Expression of CRFR1receptor-positive cell ratehave been increased after corticotropin releasing factor（CRF）acting on thelines U87. There was a significant statistically difference. The expression ofCRFR1receptor-positive cell rate were influenced attributed to concentrationof CRF and action time. Concentration and time of CRF may equally to theresults during the process. In CRF10-9mol/L for24hours, expression ofCRFR1-positive cells rate is the highest and the color is more obvious. InCRF10-9mol/L acting on U87cells, from24to48hours, raise the expressionof CRFR1receptors. But this phenomenon was not found in otherconcentration and time point，when CRF10-9mol/L acting on U87cells after12hours showing down-regulation the expression of CRFR1receptors.According to our research results, CRF influence on glioma cells byCRFR1-receptors in vitro experiments.
目的：探讨人胶质瘤细胞株U87中CRFR1受体表达及其应用不同浓度CRF作用U87细胞后CRFR1受体的表达情况。
方法：
1从培养的U87细胞中提取总RNA（ribonucleic acid），设计CRFR1引物，进行RT-PCR扩增，凝胶电泳分析结果。
2不同浓度CRF作用于胶质瘤细胞U87作用。将U87传代后随机分为四组，每组分别加入不同浓度的CRF（10-7,10-9,10-12mol/L）和不加CRF（对照组）的DMEM（含10%牛清+青链霉素）培养基，在有盖玻片的六孔培养板中继续培养U87细胞。培养12h后，每个浓度分别各取出两个孔中的细胞，进行细胞的免疫化学染色。剩余孔中细胞继续培养，在24h后以同样的方法再取出一批进行细胞免疫组化。最后一批于48h后取出行细胞免疫组化。实验重复三次。最后以细胞免疫组化结果中CRFR1表达的阳性率为结果进行统计学分析。
结果：
1RT-PCR的结果显示人胶质瘤U87中存在CRFR1mRNA表达。2.进行免疫细胞化学染色，结果显示所有组中均表达CRFR1阳性，阳性反应物位于胞浆，在细胞核周边呈现清晰淡黄色或棕黄色。以CRFR1表达的阳性率为统计数据：
2.1采用单因素方差分析不同CRF浓度对CRFR1表达的影响,每个实验组和对照组相比较，经统计学处理结果如下：12h（F=8.099，P<0.05）；24h（F=51.665，P<0.05）；48h（F=14.531，P<0.05）。可以认为不同CRF浓度对U87作用对CRFR1阳性表达率存在影响。在作用12或48h，CRF10-9mol/L和其它各浓度组比较均有差别，而其它各浓度组间比较均无差别。在作用24小时，对照组和CRF10-9mol/L比较有差别；对照组和CRF10-7mol/L，CRF10-12mol/L比较无差别; CRF10-7mol/L和CRF10-12mol/L组比较有差别。
2.2采用单因素方差分析分析不同作用时间对CRFR1表达阳性率的影响，经统计学处理结果如下：CRF10-7mol/L作用不同的时间对CRFR1表达阳性率无影响（F=1.839，P=0.238）；CRF10-12mol/L作用不同的时间对CRFR1表达阳性率无影响(F=3.884,P=0.083）；CRF10-9mol/L作用不同的时间对CRFR1表达阳性率有影响（F=90.66,P=0.0001），不同时间的多重比较中任意两组比较均有差别。
2.3对不同浓度CRF及其在不同的时间点相互交叉作用U87后CRFR1表达的阳性率数据进行析因分析；CRF不同的浓度对其CRFR1表达率有影响（F=21.185,P=0.0001）;不同的作用时间点对CRFR1表达率也有影响（F=27.954，P=0.0001）;并且不同的CRF浓度和不同的作用时间存在有交互作用（F=27.458，P=0.0001）；不同CRF药物浓度及不同的时间，CRFR1表达阳性细胞百分率有所差异，在31%～93%之间。并且表达强度亦有差别。在CRF10-9mol/L且24小时，结果显示为CRFR1表达阳性最强，呈棕褐色，并且表达阳性率最高，平均在87%左右。在CRF10-9mol/L作用48小时，CRFR1表达阳性率也较高，平均在68%左右。
结论：在RT-PCR结果中人胶质瘤U87中表达CRFR1mRNA。 CRF作用于胶质瘤U87细胞后，其CRFR1受体表达的阳性细胞率在统计学上有意义。CRF的浓度及其作用时间均可影响其CRFR1受体细胞阳性率的表达，并且不同浓度和时间存在交互作用。在CRF为10-9mol/L作用24小时时，CRFR1阳性细胞表达率最高，染色更明显。在CRF10-9mol/L时作用U87细胞，在24和48小时间上调了CRFR1受体的表达。但在其它浓度和时间点是这种现象则不明显，在CRF10-9mol/L时12小时表现为对CRFR1受体的下调。据我们的研究结果，推测CRF通过其受体CRFR1的表达对体外胶质瘤细胞的生长确实起到了一定影响的作用。
Corticotropin releasing factor（CRF）is an important neuroendocrinepeptide, and the main physiological function is to stimulate the pituitary glandsynthesis and secretion of adrenocorticotropic hormon（ACTH）and relatedpeptide. CRF and its peptide family common regulate the function ofhypothalamus-pituitary-adrenal (HPA) axis and coordinate the endocrinesystem, the autonomic nervous system, immune system, physiology andbehavior function when the body under stress. And, it has a moderating effecton cardiovascular, gastrointestinal, reproductive, skin, etc. CRF and its relatedpeptide play a role through its receptor（Corticotropin releasing factorreceptor，CRFR）. CRF receptors play a role through activation of adenylatecyclase, which belong to the G-protein coupling receptors. As the mainreceptor in the HPA axis, corticotropin releasing hormone receptor1(CRFR1)can promote the release of the pituitary ACTH to mediated CRF by the classicway under stress response. Overexpression of CRFR1recepor has also beenfound in so many human malignant tumors in the central nervous system.However, previous studies have not investigated the relationship of CRF andgliomas.
Objective：To explored the expression of CRFR1and the effcet of CRFinducing CRFR1in glioma cell line U87.
Methods：
1The total RNA were extracted from cultured U87cells. Primer of CRFR1were designed for the RT-PCR amplification. The product was analysis by gelelectrophoresis.
2Glioma cell lines U87were treated by different concentrations of CRF. U87cells was divided into four groups, which be dealed with by differentconcentrations of CRF (10-7,10-9,10-12mol/L) and control groups. Then U87 cells were cultured continuely in filled with coverslip of tin plates. Each twoholes were taken out to further immunostainned after12hours. The cells ofsurplus holes continue to culture. Another batch of cells were treated in thesame way after24hours. The last ones were treated in the same way after48hours. The experiment was repeated for three times. Finally, expression ofCRFR1positive rate were carried out by statistical analysis.
Results：
1CRFR1mRNA was expressed in human glioma cells U87by RT-PCR.2.CRFR1positive expression can be shown in every group byimmunocytochemistry. Positive protein located in the cytoplasm, andpresented a clear yellow or brown stain in the periphery of the nucleus.CRFR1-positive expression rates were carried out by statistically analysis.
2.1The single factor analysis were used for different concentration effect onthe expression the CRFR1. Compared with the control groups，differentconcentrations showed significant difference, which including12hours(F=8.099, P<0.05);24hours（F=51.665，P<0.05）;48hours (F=14.531,P<0.05). Compare the CRF10-9mol/L group with other concentration ones,there were difference at24hours and48hours. While compare with otherconcentration groups, there is no difference. At24hours, the control groupand CRF10-9mol/L group is significant difference. However, there is nodifference among the control group，CRF10-7mol/L group and CRF10-12mol/Lones. There is significant difference between the CRF10-7mol/L groups andCRF10-12mol/L ones.
2.2The single factor analysis were used for different time effect on theexpression the CRFR1. The statistics results are as follows：there is noinfluence to CRFR1expression positive rate in different times byCRF10-7mol/L treated（F=1.839，P=0.238）;and there is no influence to CRFR1expression positive rate in different times by CRF10-12mol/L treated（F=3.884,P=0.083）；there have influence to CRFR1expression positive ratein different times byCRF10-9mol/Ltreated(F=90.66,P=0.0001）, multiplecomparisons of the different time points, any two groups of comparisons are differences.
2.3Factorial analysis were carried out for different concentrations of CRFand different time to inducing CRFR1positive-expression rate. CRFR1expression rate were inducing by different concentration of CRF（F=21.185,P=0.0001）. CRFR1expression rate were influenced by different time（F=27.954, P=0.0001）; And, there are interaction between differentconcentration of CRF and different time（F=27.458, P=0.0001）. CRFR1expression rate were influenced by different concentration and time of CRF,which about31percent to93percent. And expression intensity was alsodifferences in conclusion. In CRF10-9mol/L and24hours, and the resultsshowed that CRFR1expression was strongest (the brown color)， andexpression of the highest positive rate,87%on average. In CRF10-9mol/L and48hours，expression of CRFR1-positive rate were68%on average.
Conclusion：The expression of CRFR1mRNA was found by RT-PCR inhuman glioma cells U87. Expression of CRFR1receptor-positive cell ratehave been increased after corticotropin releasing factor（CRF）acting on thelines U87. There was a significant statistically difference. The expression ofCRFR1receptor-positive cell rate were influenced attributed to concentrationof CRF and action time. Concentration and time of CRF may equally to theresults during the process. In CRF10-9mol/L for24hours, expression ofCRFR1-positive cells rate is the highest and the color is more obvious. InCRF10-9mol/L acting on U87cells, from24to48hours, raise the expressionof CRFR1receptors. But this phenomenon was not found in otherconcentration and time point，when CRF10-9mol/L acting on U87cells after12hours showing down-regulation the expression of CRFR1receptors.According to our research results, CRF influence on glioma cells byCRFR1-receptors in vitro experiments.
引文
1Fogarty DJ,Sanchez-Gomez MV,Matute C,et al. Multiple angiotensinreceptor subtypes in normal and tumor astrocytes in vitro.Glia,2002Sep；39(3):304-13
2Friend KE,Khandwala HM,Flyvbjerg A,et al. Growth hormone andinsulin-like growth factor-I: effects on the growth of glioma cell lines.Growth Horm IGF Res.2001Apr；11(2):84-91
3Sharma A,Walter J,Gozes Y,et al. A vasoactive intestinal peptideantagonist inhibits the growth of glioblastoma cells. J Mol Neurosci.2001Dec；17(3):331-9
4Kucharczak J,Pannequin J,Camby I,et al. Gastrin induces over-expressionof genes involved in human U373glioblastoma cell migration.Oncogene.2001Oct25;20(48):7021-8
5Graziani G, Tentori L, Portarena I,et al. CRF inhibits cell growth of humanendometrial adenocarcinoma cells via CRF-receptor1-mediated activationof cAMP-PKA pathway. Endocrinology.2002;143:807-813
6Carlson KW, Nawy SS, Wei ET,et al. Inhibition of mouse melanoma cellproliferation by corticotropin-releasing hormone and its analogs.Anticancer Res.2001;21:1173-1179
7Graziani G, Tentori L, Muzi A,et al. Evidence that corticotropin-releasinghormone inhibits cell growth of human breast cancer cells via theactivation of CRF-R1receptor subtype. Mol Cell Endocrinol.2007;264:44-49
8Slominski AT, Roloff B, Zbytek B,et al. Corticotropin releasing hormoneand related peptides can act as bioregulatory factors in humankeratinocytes. In Vitro Cell Dev Biol Anim.2000;36:211-216
9Dermitzaki E, Tsatsanis C, Gravanis A,et al. Corticotropin-releasinghormone induces Fas ligand production and apoptosis in PC12cells viaactivation of p38mitogen-activated protein kinase. J Biol Chem.2002;277:122801-2287
10Radulovic M, Hippel C, Spiess J. Corticotropin-releasing factor (CRF)rapidly suppresses apoptosis by acting upstream of the activation ofcaspases. J Neurochem.2003;84:10741-085
11Minas V, Rolaki A, Kalantaridou SN,et al. Intratumoral CRF modulatesimmuno-escape of ovarian cancer cells through FasL regulation. Br JCancer.2007;97:6376-45
12Kallergi G, Agelaki S, Markomanolaki H,et al. Activation of FAK/PI3K/Rac1signaling controls actin reorganization and inhibits cellmotility in human cancer cells. Cell Physiol Biochem.2007;20:977-986
13Tjuvajev J, Kolesnikov Y, Joshi R, et al. Anti-neoplastic properties ofhuman corticotropin releasing factor: involvement of the nitric oxidepathway. In Vivo,1998,12:1-10
14Ha BK,Bishop GA,King JS,et al. Corticotropin releasing factor inducesproliferation of cerebellar astrocytes. J Neurosci Res.2000Dec15；62(6):789-98
15Wang W, Ji P, Riopelle RJ, et al. Functional expression of corticotropin-releasing hormone (CRF) receptor1in cultured rat microglia. J Neurochem,2002,80:287
16Du F,Yin L,Shi M, et al. Involvement of microglial cells in infrasonicnoise-induced stress via upregulated expression of corticotrophin releasinghormone type1receptor.Neuroscience.2010May19;167(3):909-19
17Reubi JC, Waster B, Vale W, et al. Expression of CRF1and CRF2rec-eptors in human cancers. J Clin Endocrinol&Metab,2003,88(7):3312-3320
18Bayatti N, Zschocke J, Behl C. Brain region-specific neuroprotectiveaction and signaling of corticotropin-releasing hormone in primaryneurons. Endocrinology,2003,144(9):4051-60
19Alicia Arranz, MariaVenihaki, Berber Mol, et al. The impact of stress ontumor growth: peripheral CRF mediates tumor-promoting effects of stress[J].Mol Cancer.2010;9:261
20Zhang Y,Xing X,Zhan H et al. EGFR inhibitor enhances cisplatinsensitivity of human glioma cells.J Huazhong Univ Sci Technolog MedSci.2011Dec;31(6):773-8
21Moroz MA,Huang R,Kochetkow T, et al. Comparison of corticotropin-releasing factor, dexamethasone, and temozolomide: treatment efficacyand toxicity in U87and C6intracranial gliomas.Clin Cancer Res.2011May15;17(10):3282-92
22Graziani G, Tentori L, Portarena I, et al: Corticotropin-releasing hormoneinhibits cell growth of human endometrial adenocarcinoma cells viaCRH-receptor1-mediated activation of cAMP-protein kinase A pathway.Endocrinology2002；143:807-813
23Grammatopoulos DK and Chrousos GP. Functional characteristics of CRH
receptors and potential clinical applications of CRH-receptor antagonists.
Trends Endocrinol Metab.2002Dec;13:436-444
1Vale W, Spiess J, Rivier C, et al. Characterization of a41-residue ovinehypo thalamic peptide that stimulates secretion of corticotropin an β-endorphin. Science,1981,213:1394-1397
2Keck M E, H olsboer F. Hyper activity of CRF neuronal circuits as a targetfor therapeutic interventions in affective disorders [J]. Peptides,2001,22(5):835-844
3Donaldson CJ, Sutton SW, Perrin MH, et al. Cloning and characterizationof human Urocortin. Endocrinology,1996,137:2167-2170
4Vasconcelos L A, Donaldson C, Sita L V, et al. Urocortin in the centralnervous system of a primate(Cebusapella):sequencing, immunohisto-chemical, and hybridization histochemical characterization[J]. J CompNeurol,2003,463(2):157-175
5Orth D N, Mount C D. Specific high-affinity binding protein for humancorticotrophin-releasing hormone in normal human plasma[J]. BiochemBiophys Res Comm,1987,143(2):411-417
6Zouboulis C C, Sel tmann H, Hi roi N, et al. Corticotropin-r eleasinghormone: an autocrine hormone that promotes lipo-genesis in humansebocytes [J]. Proc Nat l Acad Sci U S A,2002,99(10):7148-7153
7Bowm an M E, Lopat a A, Jaff e R B, et al. Corticotropin r e leasinghormone binding protein in primates[J]. Am J Primat ol,2001,53(3):123-130
8Cort right D N, Nicol et t i A, S easholt z A F. Molecular and bio chemicalcharacterization of the mouse brain corticotrophin releasing hormonebinding protein [J]. Mol Cell Endocrinol,1995,111(2):147-157
9Florio P, Imperat ore A, Sans everin oa F, et al. The measure ment ofmaternalplasm a corticotrophin releasing fact or (CRF)and CRF bindingprotein improves the earl y predict ion of pre eclampsia[J]. J ClinEndocrinol Met ab,2004,89(9):4673-4677
10Olymeropoulos MH, Torres R, Yanovski JA, et al. The human corticot-ropin-releasing factor receptor (CRFR) gene maps to chromosome17q12-q22. Genomics,1995,28:123-124
11Hen R, Lewis KA, Perrin MH, et al. Expression cloning of a humancorticotropin-releasing factor receptor. Proc Natl Acad SciUSA,1993,90:8967-8971
12Hauger RL, Dautzenberg FM. Regulation of the stress response bycorticotropin releasing factor receptors. In Neuroendocrinology in Phy-siology and Medicine (Conn PM, Freedman ME, eds). Humana Press,1999, pp.261-286
13Autzenberg FM, Higelin J, Teichert U. Functional characterization ofcorticotropin-releasing factor type1receptor endogenously expressed inhuman embryonic kidney293cells. Eur J Pharmacol,2000,390:51-59
14Porcher C, Juhem A, Peinnequin A, et al. Expression and effects ofmetabotropic CRF1and CRF2receptors in rat small intestine[J]. Am JPhysiol Gastrointest Liver Physiol,2005May;288(5): G1091-103
15Papadopoulou N, Chen J, Randeva HS, et al. Protein kinase A inducednegative regulation of the corticotropin-releasing in hormone R1alphareceptor-extracellularly regulated kinase signal transduction pathway: thecritical role of Ser301for signaling switchand selectivity. Mol Endocrinol,2004,18:624-639
16Punn A, Levine MA, Grammatopoulos DK. Identification of signalingmolecules mediating corticotropin-releasing hormone-R1alpha-mitogen-activated protein kinase (MAPK) interactions: the critical role of phos-phatidylinositol3-kinase inregulating ERK1/2but not p38MAPKactivation. Mol Endo-crinol,2006,20:3179-3195
17almers DT, Lovenbery TW, De Souza EB. Localization of novelcorticotropin-releasing factor receptor (CRF2) mRNA expression tospecific subcortical nuclei in rat brain: comparison with CRF1receptormRNA expression. J Neurosci,1995,15:6340-6350
18Sanchez MM, Young LJ, Plotsky PM, et al. Autoradiographic and in situhybridization localization of corticotropin-releasing factor1and2receptors in nonhuman primate brain. J Comp Neurol,1999,408:365-377
19ageyama K, Bradbury MJ, Zhao L, et al. Urocortin messenger ribonucl-eic acid, tissue distribution in the rat and regulation in thymus bylipopolysaccharide and glucocorticoids. Endocrinology,1999,140(12):5651-5658
20Palchaudhuri MR, Wille S, Mevenkamp G, et al. Corticotropin relea-sing factor receptor type1from Tupaia belangeri: cloning, functionalexpression and tissue distribution. Eur J Biochem,1998,258:78-84
21Monnikes H, Schmidt BG, Tache Y. Psychological stress inducedaccelerated colonic transit in rats involves hypothalamic CRF[J]. Gastroenterology,1993Mar；104(3):716-23
22MartinezV, Rivier J, Tache Y. Peripheral injection of a new cortico tropinreleasing factor (CRF) antagonist, astressin, blocks peripheral CRF2andabdominal surgery induced delayed gastric emptying in rats [J]. JPharmacol Exp Ther,1999Aug;290(2):629-34
23Karolyi IJ, Burrows HL,Ramesh TM,et al. Altered anxiety and weigh tgain in corticotrop in-releasing hormone-binding protein-deficient m ice[J]. Proceedings of the National Academy of Science of the Un ted Statesof America,1999Sep28;96(20):11595-600
24Bartanusz V, Jezova D,Bertini LT,et al. Stress-induced increaseinvasopressin and corticortropin-releasing factor expression in hypophysiotropic paravent ricular neurons[J]. Endocrinology,1993,132(2):895-902
25Lu L, Liu D, Ceng X. Corticotrop in-releasing factor receptor type1mediates stress-induced relapse to cocaine-conditioned p lace preferencein rats [J]. European Journal of Pharmacology,2001,415(2-3):203-208
26Bale TL, ValeWW. CRF and CRF receptors: role in stress responsivity andother behaviors[J]. Annu Rev Pharm acol Toxicol,2004,44:525-557
27Grammatopoulos DK,Chrousos GS. Functional characteristics of CRFreceptors and potential clinical applications of CRF-receptor antagonists.Trends Endocrinol Metab.2002Dec;13(10):436-44
28Sun K, Yang K, Challis JRG. Differential regulation of11beta-hydroxysteroid dehydrogenase type1and2by nitric oxide incultured human p lacental trophoblast and chorionic cell p reparation [J]1JClin EndocrinolMetab,1997,138(11):4912
29Bale TL, ValeWW. CRF and CRF receptor: role in stress responsivity andother behaviors [J]. Annu Rev Pharmacol Toxicol,2004,44:525-57
30Bale T L, Contarino A, Sm ith G W, et al. Mice deficient forcorticotrophin releasing horone receptor2display anxiety like behaviorand are hypersensitive to stress [J]. Nat Genet,2000,24(4):410-414
31Bale TL, Hoshijima M, D alton N, et a.l The cardiovascular physiologicactions of urocortin II: acute effects inmurine heart failure[J].Proc NatlAcad Sci USA,2004,101(10):3697-3702
32Huang E J, Reichardt LF. Trk receptors: roles in neuronal signaltransduction[J]. Annu Rev Biochem,2003,72:609-42
33Arai MA, Assil IQ, Abou-Samra AB. Characterization of threecorticotropin-releasing factor receptors in catfish: a novel third receptor ispredominantly expressed in pituitary and urophysis. Endocrinology,2001,142:446-454
34Reubi JC, Waster B, Vale W, et al. Expression of CRF1and CRF2rec-eptors in human cancers. J Clin Endocrinol&Metab,2003,88(7):3312-3320
35Minas V, Rolaki A, Kalantaridou SN, et al. Intratumoral CRF modulatesimmuno-escape of ovarian cancer cells through FasL regulation. Br JCancer,2007,97:637-645
36Tjuvajev J, Kolesnikov Y, Joshi R, et al. Anti-neoplastic properties ofhuman corticotropin releasing factor: involvement of the nitric oxidepathway. In Vivo,1998,12:1-10
37Ariadne Androulidaki, Erini Dermitzaki, Maria Venihaki,et al.Corticotropin Releasing Factor promotes breast cancer cell motility andinvasiveness[J] Mol Cancer.2009Jun2;8:30
38Alicia Arranz, MariaVenihaki, Berber Mol, et al. The impact of stress ontumor growth: peripheral CRF mediates tumor-promoting effects of stress[J].Mol Cancer.2010Sep27;9:261
39Mark O. Huising, Talitha van der Meulen, et al. CRFR1is expressed onpancreatic β cells, promotes β cell proliferation, and potentiates insulinsecretion in a glucose-dependent manner [J].Proc Natl Acad Sci U S A.
2010January12;107(2):912-917
40刘玉昆,张建平,陈立斌等;促肾上腺皮质激素释放激素对JAR细胞凋亡相关蛋白配体表达的影响；生殖医学杂志[J]2006，12月第15卷第6期
41范军强,赵勇刚,范波等.蛋白酶体抑制剂MG-132对体外人U87胶质瘤细胞增殖和凋亡的影响；河南科技大学学报[J].2009，Sep27（3）：161-163
42Orth DN.Corticotropin releasing hormone in human. Endocri Rev,1992,13:164
43Kim Y,Park MK,Chung S, et al. Protective effect of urocortinon1-methyl-4-phenylpyridinium-induced dopaminergic neuronal death.MolCells.2010Nov;30(5):427-33
44Kagevama K,Hasegawa G,Akimoto K, et al. Differential regulation ofgonadotropin-releasing hormone by corticotropin-releasing factor familypeptides in hypothalamic N39cells.Peptides.2012Jan;33(1):149-55
45Mastorakos G, Scopa CD, Vryonidou A, et al. Presence of immunorea-ctive corticotropin-relasing hormone in normal and polycystic ovaries.J.Clin. End ocrinol. Me tab.1994,79:1191-1197
46Calogero, A. E., N. Burrello, P. Negr-i Cesi, et al. Effect s of corticotropin-relasing hormone on ovarian estrogen product ion in vitro.Endocrinology1996，137:4161-4166
47Coste S C, K esterson R A, H eldw einK A, et al Abnormal adap tations tostress and impaired cardiovascular function in m ice lac king corticotrophin releasing hormone receptor2[J]. Nat Genet,2000,24(4):403-9
48Mastorakos, G, E. L. Webster, T. C. Friedman，et al. Immunoreactivecorticotropin-releasing hormone and it s binding sites in the rat ovary. J.Cl in. I nvest.1993，92:961-968
49Radulovic M, Hippel C, Spiess J. Corticotropin-releasing factor (CRF)rapidly suppresses apoptosis by acting upstream of the activation ofcaspases. J Neurochem.2003;84:1074-1085
50Slominski AT, Roloff B, Zbytek B, et al.Corticotropin releasing hormoneand related peptides can act as bioregulatory factors in humankeratinocytes. In Vitro Cell Dev Biol Anim.2000;36:211-216
51Graziani G, Ferrandina G, Pozzoli G, et al. Corticotropin-releasing hor-mone receptor-1in human endometrial cancer. Oncol Rep,2006,15:375-379
52Chen CY，Million M，Adelson DW，et a1．Intracistemal urecortin inhibitsvagally stimulated gastricmotility in rats：role of CRF(2)[J]. JPharmacol,2002,136(2):237-247
53Dalenda C, Ines N, Fathia B, e t al. Two medical abortion regimens forlate first trimester termination of pregnancy: a prospective randomizedtrial [J]. Contraception,2010,81(4):323-327
54Maillot C, Million M, Wei JY, et al. Peripheral corticotrop in2relea2singfactor and stress2stimulated colonic motor activity involve type1recep torin rats[J]. Gastroenterology,2000,119(6):1569-1579
55Graziani G, Tentori L, Portarena I,et al. Corticotropin-releasing hormoneinhibits cell growth of human endometrial adenocarcinoma cells viaCRH-receptor1-mediated activation of cAMP-protein kinase A pathway.Endocrinology2002;143:807-813
2Friend KE,Khandwala HM,Flyvbjerg A,et al. Growth hormone andinsulin-like growth factor-I: effects on the growth of glioma cell lines.Growth Horm IGF Res.2001Apr；11(2):84-91
3Sharma A,Walter J,Gozes Y,et al. A vasoactive intestinal peptideantagonist inhibits the growth of glioblastoma cells. J Mol Neurosci.2001Dec；17(3):331-9
4Kucharczak J,Pannequin J,Camby I,et al. Gastrin induces over-expressionof genes involved in human U373glioblastoma cell migration.Oncogene.2001Oct25;20(48):7021-8
5Graziani G, Tentori L, Portarena I,et al. CRF inhibits cell growth of humanendometrial adenocarcinoma cells via CRF-receptor1-mediated activationof cAMP-PKA pathway. Endocrinology.2002;143:807-813
6Carlson KW, Nawy SS, Wei ET,et al. Inhibition of mouse melanoma cellproliferation by corticotropin-releasing hormone and its analogs.Anticancer Res.2001;21:1173-1179
7Graziani G, Tentori L, Muzi A,et al. Evidence that corticotropin-releasinghormone inhibits cell growth of human breast cancer cells via theactivation of CRF-R1receptor subtype. Mol Cell Endocrinol.2007;264:44-49
8Slominski AT, Roloff B, Zbytek B,et al. Corticotropin releasing hormoneand related peptides can act as bioregulatory factors in humankeratinocytes. In Vitro Cell Dev Biol Anim.2000;36:211-216
9Dermitzaki E, Tsatsanis C, Gravanis A,et al. Corticotropin-releasinghormone induces Fas ligand production and apoptosis in PC12cells viaactivation of p38mitogen-activated protein kinase. J Biol Chem.2002;277:122801-2287
10Radulovic M, Hippel C, Spiess J. Corticotropin-releasing factor (CRF)rapidly suppresses apoptosis by acting upstream of the activation ofcaspases. J Neurochem.2003;84:10741-085
11Minas V, Rolaki A, Kalantaridou SN,et al. Intratumoral CRF modulatesimmuno-escape of ovarian cancer cells through FasL regulation. Br JCancer.2007;97:6376-45
12Kallergi G, Agelaki S, Markomanolaki H,et al. Activation of FAK/PI3K/Rac1signaling controls actin reorganization and inhibits cellmotility in human cancer cells. Cell Physiol Biochem.2007;20:977-986
13Tjuvajev J, Kolesnikov Y, Joshi R, et al. Anti-neoplastic properties ofhuman corticotropin releasing factor: involvement of the nitric oxidepathway. In Vivo,1998,12:1-10
14Ha BK,Bishop GA,King JS,et al. Corticotropin releasing factor inducesproliferation of cerebellar astrocytes. J Neurosci Res.2000Dec15；62(6):789-98
15Wang W, Ji P, Riopelle RJ, et al. Functional expression of corticotropin-releasing hormone (CRF) receptor1in cultured rat microglia. J Neurochem,2002,80:287
16Du F,Yin L,Shi M, et al. Involvement of microglial cells in infrasonicnoise-induced stress via upregulated expression of corticotrophin releasinghormone type1receptor.Neuroscience.2010May19;167(3):909-19
17Reubi JC, Waster B, Vale W, et al. Expression of CRF1and CRF2rec-eptors in human cancers. J Clin Endocrinol&Metab,2003,88(7):3312-3320
18Bayatti N, Zschocke J, Behl C. Brain region-specific neuroprotectiveaction and signaling of corticotropin-releasing hormone in primaryneurons. Endocrinology,2003,144(9):4051-60
19Alicia Arranz, MariaVenihaki, Berber Mol, et al. The impact of stress ontumor growth: peripheral CRF mediates tumor-promoting effects of stress[J].Mol Cancer.2010;9:261
20Zhang Y,Xing X,Zhan H et al. EGFR inhibitor enhances cisplatinsensitivity of human glioma cells.J Huazhong Univ Sci Technolog MedSci.2011Dec;31(6):773-8
21Moroz MA,Huang R,Kochetkow T, et al. Comparison of corticotropin-releasing factor, dexamethasone, and temozolomide: treatment efficacyand toxicity in U87and C6intracranial gliomas.Clin Cancer Res.2011May15;17(10):3282-92
22Graziani G, Tentori L, Portarena I, et al: Corticotropin-releasing hormoneinhibits cell growth of human endometrial adenocarcinoma cells viaCRH-receptor1-mediated activation of cAMP-protein kinase A pathway.Endocrinology2002；143:807-813
23Grammatopoulos DK and Chrousos GP. Functional characteristics of CRH
receptors and potential clinical applications of CRH-receptor antagonists.
Trends Endocrinol Metab.2002Dec;13:436-444
1Vale W, Spiess J, Rivier C, et al. Characterization of a41-residue ovinehypo thalamic peptide that stimulates secretion of corticotropin an β-endorphin. Science,1981,213:1394-1397
2Keck M E, H olsboer F. Hyper activity of CRF neuronal circuits as a targetfor therapeutic interventions in affective disorders [J]. Peptides,2001,22(5):835-844
3Donaldson CJ, Sutton SW, Perrin MH, et al. Cloning and characterizationof human Urocortin. Endocrinology,1996,137:2167-2170
4Vasconcelos L A, Donaldson C, Sita L V, et al. Urocortin in the centralnervous system of a primate(Cebusapella):sequencing, immunohisto-chemical, and hybridization histochemical characterization[J]. J CompNeurol,2003,463(2):157-175
5Orth D N, Mount C D. Specific high-affinity binding protein for humancorticotrophin-releasing hormone in normal human plasma[J]. BiochemBiophys Res Comm,1987,143(2):411-417
6Zouboulis C C, Sel tmann H, Hi roi N, et al. Corticotropin-r eleasinghormone: an autocrine hormone that promotes lipo-genesis in humansebocytes [J]. Proc Nat l Acad Sci U S A,2002,99(10):7148-7153
7Bowm an M E, Lopat a A, Jaff e R B, et al. Corticotropin r e leasinghormone binding protein in primates[J]. Am J Primat ol,2001,53(3):123-130
8Cort right D N, Nicol et t i A, S easholt z A F. Molecular and bio chemicalcharacterization of the mouse brain corticotrophin releasing hormonebinding protein [J]. Mol Cell Endocrinol,1995,111(2):147-157
9Florio P, Imperat ore A, Sans everin oa F, et al. The measure ment ofmaternalplasm a corticotrophin releasing fact or (CRF)and CRF bindingprotein improves the earl y predict ion of pre eclampsia[J]. J ClinEndocrinol Met ab,2004,89(9):4673-4677
10Olymeropoulos MH, Torres R, Yanovski JA, et al. The human corticot-ropin-releasing factor receptor (CRFR) gene maps to chromosome17q12-q22. Genomics,1995,28:123-124
11Hen R, Lewis KA, Perrin MH, et al. Expression cloning of a humancorticotropin-releasing factor receptor. Proc Natl Acad SciUSA,1993,90:8967-8971
12Hauger RL, Dautzenberg FM. Regulation of the stress response bycorticotropin releasing factor receptors. In Neuroendocrinology in Phy-siology and Medicine (Conn PM, Freedman ME, eds). Humana Press,1999, pp.261-286
13Autzenberg FM, Higelin J, Teichert U. Functional characterization ofcorticotropin-releasing factor type1receptor endogenously expressed inhuman embryonic kidney293cells. Eur J Pharmacol,2000,390:51-59
14Porcher C, Juhem A, Peinnequin A, et al. Expression and effects ofmetabotropic CRF1and CRF2receptors in rat small intestine[J]. Am JPhysiol Gastrointest Liver Physiol,2005May;288(5): G1091-103
15Papadopoulou N, Chen J, Randeva HS, et al. Protein kinase A inducednegative regulation of the corticotropin-releasing in hormone R1alphareceptor-extracellularly regulated kinase signal transduction pathway: thecritical role of Ser301for signaling switchand selectivity. Mol Endocrinol,2004,18:624-639
16Punn A, Levine MA, Grammatopoulos DK. Identification of signalingmolecules mediating corticotropin-releasing hormone-R1alpha-mitogen-activated protein kinase (MAPK) interactions: the critical role of phos-phatidylinositol3-kinase inregulating ERK1/2but not p38MAPKactivation. Mol Endo-crinol,2006,20:3179-3195
17almers DT, Lovenbery TW, De Souza EB. Localization of novelcorticotropin-releasing factor receptor (CRF2) mRNA expression tospecific subcortical nuclei in rat brain: comparison with CRF1receptormRNA expression. J Neurosci,1995,15:6340-6350
18Sanchez MM, Young LJ, Plotsky PM, et al. Autoradiographic and in situhybridization localization of corticotropin-releasing factor1and2receptors in nonhuman primate brain. J Comp Neurol,1999,408:365-377
19ageyama K, Bradbury MJ, Zhao L, et al. Urocortin messenger ribonucl-eic acid, tissue distribution in the rat and regulation in thymus bylipopolysaccharide and glucocorticoids. Endocrinology,1999,140(12):5651-5658
20Palchaudhuri MR, Wille S, Mevenkamp G, et al. Corticotropin relea-sing factor receptor type1from Tupaia belangeri: cloning, functionalexpression and tissue distribution. Eur J Biochem,1998,258:78-84
21Monnikes H, Schmidt BG, Tache Y. Psychological stress inducedaccelerated colonic transit in rats involves hypothalamic CRF[J]. Gastroenterology,1993Mar；104(3):716-23
22MartinezV, Rivier J, Tache Y. Peripheral injection of a new cortico tropinreleasing factor (CRF) antagonist, astressin, blocks peripheral CRF2andabdominal surgery induced delayed gastric emptying in rats [J]. JPharmacol Exp Ther,1999Aug;290(2):629-34
23Karolyi IJ, Burrows HL,Ramesh TM,et al. Altered anxiety and weigh tgain in corticotrop in-releasing hormone-binding protein-deficient m ice[J]. Proceedings of the National Academy of Science of the Un ted Statesof America,1999Sep28;96(20):11595-600
24Bartanusz V, Jezova D,Bertini LT,et al. Stress-induced increaseinvasopressin and corticortropin-releasing factor expression in hypophysiotropic paravent ricular neurons[J]. Endocrinology,1993,132(2):895-902
25Lu L, Liu D, Ceng X. Corticotrop in-releasing factor receptor type1mediates stress-induced relapse to cocaine-conditioned p lace preferencein rats [J]. European Journal of Pharmacology,2001,415(2-3):203-208
26Bale TL, ValeWW. CRF and CRF receptors: role in stress responsivity andother behaviors[J]. Annu Rev Pharm acol Toxicol,2004,44:525-557
27Grammatopoulos DK,Chrousos GS. Functional characteristics of CRFreceptors and potential clinical applications of CRF-receptor antagonists.Trends Endocrinol Metab.2002Dec;13(10):436-44
28Sun K, Yang K, Challis JRG. Differential regulation of11beta-hydroxysteroid dehydrogenase type1and2by nitric oxide incultured human p lacental trophoblast and chorionic cell p reparation [J]1JClin EndocrinolMetab,1997,138(11):4912
29Bale TL, ValeWW. CRF and CRF receptor: role in stress responsivity andother behaviors [J]. Annu Rev Pharmacol Toxicol,2004,44:525-57
30Bale T L, Contarino A, Sm ith G W, et al. Mice deficient forcorticotrophin releasing horone receptor2display anxiety like behaviorand are hypersensitive to stress [J]. Nat Genet,2000,24(4):410-414
31Bale TL, Hoshijima M, D alton N, et a.l The cardiovascular physiologicactions of urocortin II: acute effects inmurine heart failure[J].Proc NatlAcad Sci USA,2004,101(10):3697-3702
32Huang E J, Reichardt LF. Trk receptors: roles in neuronal signaltransduction[J]. Annu Rev Biochem,2003,72:609-42
33Arai MA, Assil IQ, Abou-Samra AB. Characterization of threecorticotropin-releasing factor receptors in catfish: a novel third receptor ispredominantly expressed in pituitary and urophysis. Endocrinology,2001,142:446-454
34Reubi JC, Waster B, Vale W, et al. Expression of CRF1and CRF2rec-eptors in human cancers. J Clin Endocrinol&Metab,2003,88(7):3312-3320
35Minas V, Rolaki A, Kalantaridou SN, et al. Intratumoral CRF modulatesimmuno-escape of ovarian cancer cells through FasL regulation. Br JCancer,2007,97:637-645
36Tjuvajev J, Kolesnikov Y, Joshi R, et al. Anti-neoplastic properties ofhuman corticotropin releasing factor: involvement of the nitric oxidepathway. In Vivo,1998,12:1-10
37Ariadne Androulidaki, Erini Dermitzaki, Maria Venihaki,et al.Corticotropin Releasing Factor promotes breast cancer cell motility andinvasiveness[J] Mol Cancer.2009Jun2;8:30
38Alicia Arranz, MariaVenihaki, Berber Mol, et al. The impact of stress ontumor growth: peripheral CRF mediates tumor-promoting effects of stress[J].Mol Cancer.2010Sep27;9:261
39Mark O. Huising, Talitha van der Meulen, et al. CRFR1is expressed onpancreatic β cells, promotes β cell proliferation, and potentiates insulinsecretion in a glucose-dependent manner [J].Proc Natl Acad Sci U S A.
2010January12;107(2):912-917
40刘玉昆,张建平,陈立斌等;促肾上腺皮质激素释放激素对JAR细胞凋亡相关蛋白配体表达的影响；生殖医学杂志[J]2006，12月第15卷第6期
41范军强,赵勇刚,范波等.蛋白酶体抑制剂MG-132对体外人U87胶质瘤细胞增殖和凋亡的影响；河南科技大学学报[J].2009，Sep27（3）：161-163
42Orth DN.Corticotropin releasing hormone in human. Endocri Rev,1992,13:164
43Kim Y,Park MK,Chung S, et al. Protective effect of urocortinon1-methyl-4-phenylpyridinium-induced dopaminergic neuronal death.MolCells.2010Nov;30(5):427-33
44Kagevama K,Hasegawa G,Akimoto K, et al. Differential regulation ofgonadotropin-releasing hormone by corticotropin-releasing factor familypeptides in hypothalamic N39cells.Peptides.2012Jan;33(1):149-55
45Mastorakos G, Scopa CD, Vryonidou A, et al. Presence of immunorea-ctive corticotropin-relasing hormone in normal and polycystic ovaries.J.Clin. End ocrinol. Me tab.1994,79:1191-1197
46Calogero, A. E., N. Burrello, P. Negr-i Cesi, et al. Effect s of corticotropin-relasing hormone on ovarian estrogen product ion in vitro.Endocrinology1996，137:4161-4166
47Coste S C, K esterson R A, H eldw einK A, et al Abnormal adap tations tostress and impaired cardiovascular function in m ice lac king corticotrophin releasing hormone receptor2[J]. Nat Genet,2000,24(4):403-9
48Mastorakos, G, E. L. Webster, T. C. Friedman，et al. Immunoreactivecorticotropin-releasing hormone and it s binding sites in the rat ovary. J.Cl in. I nvest.1993，92:961-968
49Radulovic M, Hippel C, Spiess J. Corticotropin-releasing factor (CRF)rapidly suppresses apoptosis by acting upstream of the activation ofcaspases. J Neurochem.2003;84:1074-1085
50Slominski AT, Roloff B, Zbytek B, et al.Corticotropin releasing hormoneand related peptides can act as bioregulatory factors in humankeratinocytes. In Vitro Cell Dev Biol Anim.2000;36:211-216
51Graziani G, Ferrandina G, Pozzoli G, et al. Corticotropin-releasing hor-mone receptor-1in human endometrial cancer. Oncol Rep,2006,15:375-379
52Chen CY，Million M，Adelson DW，et a1．Intracistemal urecortin inhibitsvagally stimulated gastricmotility in rats：role of CRF(2)[J]. JPharmacol,2002,136(2):237-247
53Dalenda C, Ines N, Fathia B, e t al. Two medical abortion regimens forlate first trimester termination of pregnancy: a prospective randomizedtrial [J]. Contraception,2010,81(4):323-327
54Maillot C, Million M, Wei JY, et al. Peripheral corticotrop in2relea2singfactor and stress2stimulated colonic motor activity involve type1recep torin rats[J]. Gastroenterology,2000,119(6):1569-1579
55Graziani G, Tentori L, Portarena I,et al. Corticotropin-releasing hormoneinhibits cell growth of human endometrial adenocarcinoma cells viaCRH-receptor1-mediated activation of cAMP-protein kinase A pathway.Endocrinology2002;143:807-813