益骨胶囊含药血清对成骨细胞ER和IL-6影响的研究
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摘要
目的：
运用中药血清药理学方法体外观察益骨胶囊含药血清对新生大鼠成骨细胞增殖、分化、矿化的影响，探讨制备含药血清的时效和量效关系。同时从mRNA水平及蛋白水平研究益骨胶囊含药血清对成骨细胞ER和IL-6表达的影响，试图从雌激素调控途径探讨益骨胶囊防治骨质疏松症的机理。
方法：
(1)将12月龄SD大鼠分40只，随机分为4组(益骨胶囊高、中、低剂量组和生理盐水组)，制备含药血清和空白对照血清；(2)分离培养新生SD大鼠的OB，采用MTT法、分光光度计比色法和茜素红染色法，分别测定3组即敏感浓度含药血清组(高剂量稀释比1：4)、空白灭活对照血清组和DMEM培养液组对OB增殖、分化和矿化的影响；(3)用RT-PCR技术和RBA法分别检测3组对OB表达ERαmRNA、ERβmRNA及其蛋白的影响；(4)用RT-PCR技术和RIA试剂盒分别检测3组对OBIL-6mRNA及其蛋白表达的影响。
结果：
(1)末次灌胃后1.5h采血的MTT值和同组其它时间比较具有统计学意义(P＜0.05)；血清添加浓度为25％时高剂量组MTT值最高，和其它组比较具有统计学意义(P＜0.05)。(2)益骨胶囊含药血清组ALP值及OB矿化结节数量均明显高于空白血清对照组及培养液组(P＜0.05)。(3)益骨胶囊含药血清组ERα／β-actin的灰度积分值和对照组比较无统计学意义(P＞0.05)；ERβ／β-actin高于对照组(P＜0.05)；IL-6／β-actin低于对照组(P＜0.05)。(4)益骨胶囊含药血清组KD值和对照组比较不具有统计学意义(P＞0.05)；RT值明显高于对照组(P＜0.01)。(5)益骨胶囊含药血清组IL-6含量明显低于对照组(P＜0.05)。
结论：
益骨胶囊含药血清能促进成骨细胞增殖、分化、矿化；能上调OB ERβmRNA表达，增加ER蛋白数量；能下调OB IL-6mRNA表达，抑制OB分泌IL-6；这提示益骨胶囊主要通过ERβ来发挥类雌激素作用。它能促进体外培养的成骨细胞的骨形成，同时可能也影响骨吸收。通过雌激素及其受体调控途径防治OP是益骨胶囊的药理机制之一。
Objective: To use the method of Chinese herbal seropharmacology, to investigate the effect of the rats sera containing Benefiting-bone capsule (BBC) on the cell proliferation, cell differentiation and cell mineralization of rat osteoblast in vitro skulls of neonatal SD rats and to research the relationship between the best time-effectiveness and dosage-effectiveness, and at the same time, to study the significant changes of both alpha and beta estrogen receptors in the osteoblast and the production of Interleukin-6(IL-6) by adding the rats sera containing BBC by studying the expression level of both the mRNA and the protein. This is performed to explore the mechanism of BBC on the prevention and treatment osteoporosis by means of estrogenic modulation and control.
Methods: Forty female SD rats, aged 12 months, were used and randomly divided into 4 groups including N.S., low dosage, middle dosage and high dosage with BBC, the sera were extracted at different times. The MTT method was used to determine the best time-efficiency and dosage-efficiency. To obtain osteoblasts from neonatal SD rat's skull using the method of twice enzyme digestion, conducted primary culture and subcultures.There are three groups including Group I (containing deactivating sera with BBC), Group II (containing deactivating sera without BBC) and Group III (simple DMEM medium group). Also, spectrophoto- meter colorimetry assays were used to examine the activity of the alkaline phospha tase. Alizarin at 0.1% was used to count the mineralized nodes in discussing the influence involving the differentiation and the mineralization of the osteoblast of the rats sera containing BBC. RT-PCR technology was used to study the estrogenic receptor α,
βmRNA, IL-6 mRNA and p-actin mRNA. The RBA method was u
sed to study the affinity of estrogen receptors and the number of receptors. The RIA method was used to study IL-6 in the culture (including the sera) of the osteoblasts.
Results: The MTT value reached its highest point 1.5 hours after oral administration, which was considered to be statistically significant when compared with others (P<0.05). MTT of high dosage met the maximum value with the concen tration of 25%, and was considered to be statistically significant (P<0.05). Thereby increasing the number of ALP, mineralized nodes and up-modulating the expression of ERp mRNA ( P<0.05), without boosting the affinity of ER compared to the control groups (P>0.05) but increasing the receptors number statistically significantly(P<0.01).
Both can down-modulate the expression of IL-6 mRNA and protein ( P<0.05). Conclusions: The rats sera containing BBC can promote proliferation, differentiation and mineralization of osteoblasts, there is an up-modulating of the osteoblastular ERβ mRNA and an increase in the number of their receptors' protein, at the same time, a down-modulating of the osteoblastular IL-6 mRNA and a decrease in the amount of IL-6 secreted by osteoblasts. These indicate an estrogenic action on bones through ERβ modulation. BBC, a Chinese herbal complex prescription, can effectively treat Osteoporosis through the modulating mechanism of estrogenic pathway. This is worth further research.
运用中药血清药理学方法体外观察益骨胶囊含药血清对新生大鼠成骨细胞增殖、分化、矿化的影响，探讨制备含药血清的时效和量效关系。同时从mRNA水平及蛋白水平研究益骨胶囊含药血清对成骨细胞ER和IL-6表达的影响，试图从雌激素调控途径探讨益骨胶囊防治骨质疏松症的机理。
方法：
(1)将12月龄SD大鼠分40只，随机分为4组(益骨胶囊高、中、低剂量组和生理盐水组)，制备含药血清和空白对照血清；(2)分离培养新生SD大鼠的OB，采用MTT法、分光光度计比色法和茜素红染色法，分别测定3组即敏感浓度含药血清组(高剂量稀释比1：4)、空白灭活对照血清组和DMEM培养液组对OB增殖、分化和矿化的影响；(3)用RT-PCR技术和RBA法分别检测3组对OB表达ERαmRNA、ERβmRNA及其蛋白的影响；(4)用RT-PCR技术和RIA试剂盒分别检测3组对OBIL-6mRNA及其蛋白表达的影响。
结果：
(1)末次灌胃后1.5h采血的MTT值和同组其它时间比较具有统计学意义(P＜0.05)；血清添加浓度为25％时高剂量组MTT值最高，和其它组比较具有统计学意义(P＜0.05)。(2)益骨胶囊含药血清组ALP值及OB矿化结节数量均明显高于空白血清对照组及培养液组(P＜0.05)。(3)益骨胶囊含药血清组ERα／β-actin的灰度积分值和对照组比较无统计学意义(P＞0.05)；ERβ／β-actin高于对照组(P＜0.05)；IL-6／β-actin低于对照组(P＜0.05)。(4)益骨胶囊含药血清组KD值和对照组比较不具有统计学意义(P＞0.05)；RT值明显高于对照组(P＜0.01)。(5)益骨胶囊含药血清组IL-6含量明显低于对照组(P＜0.05)。
结论：
益骨胶囊含药血清能促进成骨细胞增殖、分化、矿化；能上调OB ERβmRNA表达，增加ER蛋白数量；能下调OB IL-6mRNA表达，抑制OB分泌IL-6；这提示益骨胶囊主要通过ERβ来发挥类雌激素作用。它能促进体外培养的成骨细胞的骨形成，同时可能也影响骨吸收。通过雌激素及其受体调控途径防治OP是益骨胶囊的药理机制之一。
Objective: To use the method of Chinese herbal seropharmacology, to investigate the effect of the rats sera containing Benefiting-bone capsule (BBC) on the cell proliferation, cell differentiation and cell mineralization of rat osteoblast in vitro skulls of neonatal SD rats and to research the relationship between the best time-effectiveness and dosage-effectiveness, and at the same time, to study the significant changes of both alpha and beta estrogen receptors in the osteoblast and the production of Interleukin-6(IL-6) by adding the rats sera containing BBC by studying the expression level of both the mRNA and the protein. This is performed to explore the mechanism of BBC on the prevention and treatment osteoporosis by means of estrogenic modulation and control.
Methods: Forty female SD rats, aged 12 months, were used and randomly divided into 4 groups including N.S., low dosage, middle dosage and high dosage with BBC, the sera were extracted at different times. The MTT method was used to determine the best time-efficiency and dosage-efficiency. To obtain osteoblasts from neonatal SD rat's skull using the method of twice enzyme digestion, conducted primary culture and subcultures.There are three groups including Group I (containing deactivating sera with BBC), Group II (containing deactivating sera without BBC) and Group III (simple DMEM medium group). Also, spectrophoto- meter colorimetry assays were used to examine the activity of the alkaline phospha tase. Alizarin at 0.1% was used to count the mineralized nodes in discussing the influence involving the differentiation and the mineralization of the osteoblast of the rats sera containing BBC. RT-PCR technology was used to study the estrogenic receptor α,
βmRNA, IL-6 mRNA and p-actin mRNA. The RBA method was u
sed to study the affinity of estrogen receptors and the number of receptors. The RIA method was used to study IL-6 in the culture (including the sera) of the osteoblasts.
Results: The MTT value reached its highest point 1.5 hours after oral administration, which was considered to be statistically significant when compared with others (P<0.05). MTT of high dosage met the maximum value with the concen tration of 25%, and was considered to be statistically significant (P<0.05). Thereby increasing the number of ALP, mineralized nodes and up-modulating the expression of ERp mRNA ( P<0.05), without boosting the affinity of ER compared to the control groups (P>0.05) but increasing the receptors number statistically significantly(P<0.01).
Both can down-modulate the expression of IL-6 mRNA and protein ( P<0.05). Conclusions: The rats sera containing BBC can promote proliferation, differentiation and mineralization of osteoblasts, there is an up-modulating of the osteoblastular ERβ mRNA and an increase in the number of their receptors' protein, at the same time, a down-modulating of the osteoblastular IL-6 mRNA and a decrease in the amount of IL-6 secreted by osteoblasts. These indicate an estrogenic action on bones through ERβ modulation. BBC, a Chinese herbal complex prescription, can effectively treat Osteoporosis through the modulating mechanism of estrogenic pathway. This is worth further research.
引文
[1] 吴小涛，戴克戎，裘世静．老年与青年骨内无机及微量元素分布比较．江苏医药，2000，26(2)：105-107
[2] 杨彬．半胱氨酸组织蛋白酶抑制剂：一类新型的骨质疏松治疗药物．国外医学·药学分册，2000，27(2)：79-84
[3] Zhou S, Turgeman G, Harris SE, Leitman DC, Komm BS, Bodine PV, Gazit D. Estrogens activate bone morphogenetic protein-2 gene transcription in mouse mesenchymal stem cells. Mol. Endocrinol. 2003, 17(1): 56-66
[4] Lubahn DB, Moyer JS, Golding Ts, Couse JF, Korach KS, Smithies O. Alteration of reproductive function but not prenatal sexual development after insertional disruption of the mouse estrogen receptor gene J. Proc. Natl. Acad. Sci. USA 1993, 90:11162-11061
[5] Smith FP, Boyd J, Frank GR, Takahashi H, Cohen RM, Specker B, Williams TC, Lubahn DB, Korach KS. Estrogen resistance caused by a mutation in the estrogen -receptor gene in a man J N. Engl. Med. 1994, 331: 1056-1061
[6] Zofkova I, Zajickova K, Hill M. The estrogen receptor alpha gene determines serum androstenedione levels in postmenopausal women. Steroids 2002, 67(10):815-9
[7] Kobayashi N, Fujino T, Shirogane T, Furuta I, Kobamatsu Y, Yaegashi M, Sakuragi N, Fujimoto S. Estrogen receptor alpha polymorphism as a genetic marker for bone loss, vertebral fractures and susceptibility to estrogen. Maturitas 2002, 41 (3): 193-201
[8] Allcroft LC, Varanasi SS, Dimopoulos D, Francis RM, Datta HK. Mutational and Polymorphic Analysis of the Estradiol Receptor-a Gene in Men with Symptomatic Vertebral Fractures. Calcif. Tissue Int. 2002, 71(5):400-5
[9] Fujita M, Urano T, Horie K, Ikeda K, Tsukui T, Fukuoka H, Tsutsumi O, Ouchi Y, Inoue S. Estrogen activates cyclin-dependent kinases 4 and 6 through induction of cyclin D in rat primary osteoblasts. Biochem. Biophys. Res. Commun. 2002, 299(2): 222-8
[10] Ferrari SL, Ahn-Luong L, Garnero P, Humphries SE, Greenspan SL. Two promoter polymorphisms regulating interleukin-6 gene expression are associated with circulating levels of C-reactive protein and markers of bone resorption in postmenopausal women. J. Clin. Endocrinol. Metab. 2003, 88(1):255-9
[11] Taichman Rs, Reilly MJ. Emerson SG. Human osteoblasts support human hematopoietic progenitor cells in vitro bone marrow cultures. Blood 1996, 87(2): 518-24
[12] Kitamura H, Kawata H, Takahashi F, Higuchi Y, Furuichi T, Ohkawa H. Bone marrow neutrophilia and suppressed bone turnover in human interleukin-6 transgenic mice. A cellular relationship among hematopoietic cells, osteoblasts, and osteoclasts mediated by stromal cells in bone marrow. Am J Pathol 1995, 147(6): 1682-92
[13] Rude RK, Gruber HE, Wei LY, Frausto A. Magnesium deficiency: effect on bone and mineral metabolism in the mouse. Mills BG. Calcif Tissue Int 2003 Jan; 72(1): 32-41
[14] Morello KC, Wurz GT, De Gregorio MW. SERMs: current status and future trends. Crit Rev Oncol Hematol. 2002 Jul; 43(1): 63-76
[15] Bodine PV, Harris HA, Lyttle CR, Komm BS. Estrogenic effects of 7alpha-methyl-17alpha-ethynylestradiol: a newly discovered tibolone metabolite. Steroids 2002, 67(8): 681-6
[16] Hayashi M, Rho MC, Enomoto A, Fukami A, Kim YP, Kikuchi Y, Sunazuka T, Hirose T, Komiyama K, Omura S. Suppression of bone resorption by madindoline A, a novel nonpep- tide antagonist to gp130. Proc. Natl. Acad. Sci. U. S. A. 2002, 99(23): 14728-33
[17] Ishimi Y, Yoshida M, Wakimoto S, Wu J, Chiba H, Wang X, Takeda K, Miyaura C Genistein, a soybean isoflavone, affects bone marrow lymphopoiesis and prevents bone loss in castrated male mice. Bone 2002, 31(1): 180-5
[18] Torricelli P, Fini M, Giavaresi G, Giardino R. Human osteopenic bone-derived osteoblasts: essential amino acids treatment effects. Artif Cells Blood Substit. Immobil. Biotechnol. 2003, 31(1): 35-46
[19] 吴淑贤，李钢铁．补肾壮骨胶囊治疗中老年骨质疏松症临床观察报告．广州医药，1999，30(5)：71-72
[20] 张贵有．二仙坚骨汤治疗老年性脊椎骨质疏松所致腰背部疼痛65例报告．中国中医骨伤科杂志，1999，7(2)：28-29
[21] 邵敏，黄宏兴，庄洪．骨康防治骨质疏松拆方的初步研究．中国中医骨伤科杂志 2000，8(2)：7-11
[22] 王玲，仲强惟．浅谈补肾益肝健脾治疗绝经后骨质疏松症．天津中医，2001，18(1)：40-41
[23] 黄桂成．理气止痛汤治疗原发性骨质疏松腰背痛50例．镇江医学院学报，2000，10(3)：600-601
[24] 胡军．辨证治疗绝经期骨质疏松780例．中国民间疗法，2000，8(2)：40-41
[25] 陈维静，刘新梅，廉红真．中医辨证治疗原发性骨质疏松症(附109例报告)．中医药研究，1999，15(3)：29-31
[26] 陈大蓉，唐显著，郑坤渝，刘其芬，蔡汉中，李小木，张健．中药护骨合剂防治绝经后骨质疏松症的临床研究．中医杂志，1994，35(6)：359-361
[27] 危剑安，陈训华，李承军，张云如，程谟库，杨桂香，李平，华瑞成，李宝珠，王建平，康素军，陈玉英，刘丽华，石冰．肾虚骨痛胶囊对绝经期妇女骨密度的影响．中医杂志，1995，36(9)：548-550
[28] 梁克玉，魏玉玲，郭邦富，胡年宏，付刚．中药增骨汤序贯疗法治疗绝经后骨质疏松症—附120例临床观察．中医正骨，1999，11(1)：9-10
[29] 陈玉兰，牛占和，牛占山．中西医结合治疗绝经后妇女原发性骨质疏松症98例．中国骨质疏松杂志，1999，5(2)：44-46
[30] 谢雁鸣．老年OP研究近况．中医药动态，1994，(1)：11-13
[31] 程化奇，夏振信，殷新猷．更年平调冲剂治疗骨质疏松症的药理实验和临床初步研究．中国骨质疏松杂志，1999，5(1)：71-75
[32] 张玲娟，沈自尹，蔡定芳．淫羊藿对大鼠下丘脑垂体肾上腺轴抑制模型的影响．中医杂志，1996，37(10)：620-623
[33] 刘和娣，李恩，崔灵光，佟晓旭，王桂香，陈永春．补肾中药对不同年龄女性多指标及骨密度的影响．中国中西医结合杂志，1997，17(11)：671-672
[34] 沈培芝，李东煜，张戈，史万忠，徐宇，石印玉．补肾方防治地塞米松致雄性大鼠骨质疏松及其生化机制探讨．中国中西医结合杂志，1998，18(5)：290-292
[35] 刘和娣，李恩，刘昆．补肾中药对骨质疏松大鼠CaBP-D_9K基因及表达的影响．中国骨质疏松杂志，1996，2(3)：62-64
[36] 李青南，吴铁，谢华，黄莲芳，张志平．淫羊藿提取液对去睾丸大鼠骨代谢的影响．中草药，1993，24(12)：637-638
[37] 罗为民，符诗聪，杜宁，曲克服，李国衡．健脾补肾法防治绝经后骨质疏松症的计量学研究．中国中医骨伤科，1995，3(2)：1-4
[38] 陈坤，于世凤，史风芹，庞淑珍．黔岭藿对体外培养的破骨细胞作用的研究．中国骨质疏松杂志，1996，2(3)：59-61
[39] 崔家鹏，郑洪新，刘景峰，林庶茹，夏淑杰，刘树新．补肾中药对肾虚骨质疏松症大鼠红细胞膜PKCCa~(2+)-Mg~(2+)-ATP酶活性影响的实验研究．中国骨质疏松杂志，1997，3(3)：66-69
[40] 毛平，陆光伟，周志锦，杨美东，叶福媛．六味地黄丸对阴虚动物模型各个脏器组织元素含量的影响．中国中药杂志，1993：18(11)：690-692
[41] 张继东，郭福新，陈克忠．对衰老机理的认识．山东中医杂志，1992，11(6)：45-46
[42] 李芳芳，李恩，冬晓旭，陈永春，康文华．补肾、健脾和活血化瘀方药对去卵巢大鼠骨质疏松的比较性研究．中国骨质疏松杂志，1998，4(1)：5-9
[43] 林求诚，林朝晖．中医衰老指数与中医学年龄的初步分析．中西医结合杂志，1990，10(2)：90-92
[44] 何华．老年气虚血瘀初探．新中医，1991，23(7)；52-54
[45] 吕志平．中老年弦细脉甲皱微循环观察．山东中医学院学报，1994，18(2)：140-141
[46] 杨少锋．关于骨质疏松症与血瘀关系的探讨．湖南中医药导报，1996，2(4)：25-26
[47] 田代真一．“血清药理学”“血清药化学”——汉方药理学始药物中浓度测定新世界．TDM研究，1988，1(5)：54-57
[48] 田代真一．血清药理学血清药化学．现代东洋医学，1992，13(1)：113-117
[49] 季光，刘平，洪嘉和，刘成，徐列明．扶正化瘀方药物血清对原代培养纤维肝细胞胶原生成及分泌白蛋白功能的影响．中西医结合肝病杂志，1997，7(1)：25-28
[50] 何玉军，苏安英，柴锡庆，周双俊，悦随士，门金娥．用血清药理学方法观察中药复方制剂诱导人胃癌细胞凋亡的研究．中国综合临床，1998，14(1)：30-31
[51] 贺玉琢，高英杰，沈鸿，崔晓兰，郭淑英，姜廷良．给予桂枝汤的大鼠血清对HSV-1致CPE的影响．中国实验方剂学杂志，1999，5(4)：44-47
[52] 张侠，陈信义，王沛．血清药理学方法研究益肾健脾活血法对BEL-7402细胞增殖抑制作用．实用肿瘤杂志，1999，14(2)：87-89
[53] Iwama H, Amagayas S, Ogihara Y. Effect of shosaikoto, a Japanese and Chinese traditional herbal medical mixture, on the mitogenic activity of lipopolysaccharide: a new pharmacological testing method. J. Ethnopharmacol, 1987, 21(1): 45-53
[54] Umeda M, Amagayas S, Ogihara Y. Effect of certain herbal medicines on the biotrans-formation of arachidonic acid:a new pharmacological testing method using serum. J. Ethnopharmacol, 1988, 23(1): 91-98
[55] 王力倩，余上才，李仪奎，李悦．用血清药理学方法研究中药苦参、仙鹤草的抗肿瘤作用．中国中医药科技，1995，2(5)：19-21
[56] 张群豪，钟蓓，陈可冀，史大卓，毛节明，陈明哲．用血清药理学方法观察血府逐瘀浓缩丸对实验性动脉粥样硬化家兔主动脉平滑肌细胞增殖的影响．中国中西医结合杂志，1996，16(3)：156-159
[57] 崔晓兰，贺玉琢，高英杰，李晓芹，周爱香，田甲丽，郭淑英，富杭育，姜廷良．中药复方血清药理研究方法学探讨．中国实验方剂学杂志，1998，4(2)：13-15
[58] 刘成海，刘成，刘平，徐列明．扶正化瘀复方药物血清对大鼠肝贮脂细胞增殖及胶原合成的影响．中国实验方剂学杂志，1996，2(2)：16-19
[59] 刘成海，刘平，刘成，季光，徐列明．抗肝纤维化有效中药复方血清药理学方法探讨．中国实验方剂学杂志，1998，4(2)：16-19
[60] 季光，刘平，刘成，徐列明，洪嘉禾．扶正化瘀方药物血清对原代培养大鼠肝细胞增殖及胶原生成率的影响．中国实验方剂学杂志，1997，3(3)：20-23
[61] 王伟，史大卓，陈可冀，钟蓓．精制血府胶囊刺激缺氧缺糖心肌细胞RNA及蛋白质合成的研究．北京中医药大学学报，1997，20(2)：17-19
[62] 周云中，季克胜，孙小玉，丁炜，韩兴梅，洪敏．金荞麦片的血清药理学研究——对离体豚鼠回肠的影响．中成药，1999，21(1)：34-35
[63] 阴赦宏，董自波．应用血清药理学方法观察金荞麦片拮抗组胺引起的离体豚鼠回肠收缩作用．中国中医药科技，1999，6(3)：149-151
[64] 杨奎，沈映君，王一涛，曾南，张德波，史焱．含香薷、羌活胜湿汤和九味羌活丸血清对内生致热原产生的影响．中药药理与临床，1995，(4)：1-3
[65] 王力倩，金若敏，孙峥嵘，沈玉茹，陈长勋．复方仙人球对肿瘤细胞体外生长影响的实验观察．上海中医药杂志，1997，(4)：44-46
[66] 陈长勋，金若敏，贺劲松，李仪奎．用血清药理学实验方法观察附子的强心作用．中国中医药科技，1996，3(3)：12-14
[67] 李兰芳，金亚宏，原桂东，曹秀芳，姜廷良．当归补血汤抗自由基作用的药效动力学研究．中国实验方剂学杂志，1999，5(1)：24-26
[68] 杨奎，郭力，周明眉，姜远平，王一涛，蒲旭峰，李萍，陈槐卿，吴立志．中药血清药化学与中药血清药理学协同研究方法探讨．中药药理与临床，1998，14(4)：41-44
[69] 李勇，季晖，李萍，谢林．淫羊藿总黄酮对体外培养成骨细胞的影响．中国药科大学学报，2002，33(1)：48-50
[70] 高素强，傅得兴，张红梅．淫羊藿及其复方防治骨质疏松得研究进展．中国中药杂志，1999，24(4)：249-251
[71] 郑本端，吕国桥，罗自文．龟甲胶药理作用研究进展．时珍国医国药，2001，12(5)：463-465
[72] 孟大利，李铣．中药牛膝化学成分和药理活性的研究进展．中国药物化学杂志，2001，11(2)：121-122
[73] 周丕琪，沈霖，杜靖远，杨艳萍，谢晶，高兰，刘保平．补肾法对绝经后骨质疏松症患者IL-6、TNT-α的影响．中国中医骨伤科杂志，2000，8(4)：1-3
[74] 张荣华，丘和明．中医中药退行性骨质疏松症用药分析．中医药学报，1997，(4)：30-31
[75] 张荣华，陈可冀，陆大祥，马晓昌，侯励．补肾活血液延缓雄性大鼠增龄性骨质疏松的研究．中国病理生理杂志，2001，17(12)：1205-1207
[76] 冯伟，石印玉，沈培芝，徐宇，詹红生．MTT法分析中药含药血清对体外软骨细胞增殖影响的研究．上海中医药大学学报，2000，14(1)：43-45
[77] 吴健宇，李仪奎，符胜光，周芝兰．补阳还五汤保护氧自由基损伤血管内皮细胞的血清药理实验方法的建立．中药药理与临床，1999，15(1)：45-46
[78] Bergman RJ, Gazit D, Kahn AJ. Age related changes in osteogenic stem cells in mice. J. Bone Miner. Res. 1996, 1(1): 568-577
[79] 王力倩，李仪奎，符胜光，付惠娣．血清药理学方法研究探索．中药药理与临床，1997，13(3)：29-31
[80] 孟李，王宁生．含药血清的制备方法研究．中药新药与临床药理，1999，10(5)：290-292
[81] 周明眉，杨奎，姜远平，王一涛，陈槐卿，吴立志．中药血清药理学的方法学研究——采血时间的确定及时效关系研究．中药药理与临床，1999；15(1)：43-44
[82] 李仪奎．中药血清药理学实验方法的若干问题．中药新药与临床药理，1999；10(2)：95-98
[83] 周明眉，杨奎，姜远平，王一涛．中药血清药理学的方法学研究——反应体系中含药血清加入量的研究．中药药理与临床，1998，14(6)：43-44
[84] 崔晓兰，贺玉琢，高英杰，李晓芹．中药复方血清药理研究方法学探讨Ⅰ、Ⅱ．全国中药
研究学术讨论会论文摘要集．中国中西医结合学会中药专业委员会，1997：66-67
[85] 黄于贵，陈磊，贺师鹏．~3H-TdR掺入DNA技术的正确应用．中华医学杂志，1993，73(5)：308-309
[86] Owen TA, Aronow M, Shalhoub V, Barone LM, Wilming L, Tassinari MS, Kennedy MB, Pockwinse S, Lian JB, Stein GS. Progressive development of the rat osteoblast phenotype in vitro: reciprocal relationships in expression of genes associated with osteoblast proliferation and differentiation during form ation of the bone extracellular matrix. J.Cell Physiol. 1990, 143(3):420-430
[87] Pass-Even L, Gazit D, Bab I. Ontogenesis of ultrastructural features during osteogenic differentiation in diffusion chamber cultures of marrow cells. J Bone Mine. Res. 1993, 8(5):589-95
[88] J．萨姆布卢克D．W拉塞尔著．分子克隆实验指南 第三版．科学出版社出版 2002，第一版：597-701
[89] Asok K. Dasmahapatra, Barbara A.B. Wimpee, Amanda L. Trewin, Reinhold J. Hutz 2,3,7,8-Tetrachlorodibenzo-p-dioxin increases steady-state estrogen receptor- mRNA levels after CYP1A1 and CYP1B1 induction in rat granulosa cells in vitro. J. Molecular and Cellular Endocrinology 2001, 182:39-48
[90] Stoll S., Ko" nig S., Thanos S. Isolation of total-RNA from formalin-fixed rat retina. Journal of Neuroscience Methods, 2002, 120(1): 55-63
[91] Albright F. Postmenopausal osteoporosis. Trans Assoc Am Physic, 1940, 55:298-301.
[92] Jilka RL, Hangoc G, Girasole G, Passeri G, Williams DC, Abrams JS, Boyce B, Broxmeyer H, Manolagas SC. Science, 1992, 257 (5066): 88-91
[93] Tabibzadeh SS, Santhanam U, Sehgal PB, May LT. Cytokine-induced production of IFN-beta 2/IL-6 by freshly explanted human endometrial stromal cells. Modulation by estradiol-17 beta. J. Immunol, 1989, 142(9):3134-3139
[94] Chow J, Tobias JH, Colston KW, Chambers TJ. Estrogen maintains trabecular bone volume in rats not only by suppression of bone resorption but also by stimulation of bone formation. J Clin. Invest, 1992, 89(1):74-78
[95] Kuiper g Shughrue PJ, Merchenthaler I, Gustafsson JA. The estrogen receptor beta subtype: a novel mediator of estrogen action in neuroendocrine systems. Front Neuroendocrinol, 1998, 19(4):253-286
[96] Couse JF, Jonathan L, Grandien K, Gustafsson JA, Korach KS. Tissue distribution and quantitative analysis of estrogen receptor α(ERα)and estrogen receptor β(ERβ) messenger ribonucleic acid in the wild type and Er α knockout mouse. J. Endocrinology, 1997, 138(11): 4613-4612
[97] Onoe Y, Miyaura C, Ohta H, Nozawa S, Suda T. Expression of estrogen receptor β in rat bone. J. Endocrinology, 1997, 138(10): 4059-4512
[98] Bodine PV, Henderson RA, Green J, Aronow M, Owen T, Stein GS, Lian JB, Komm BS. Estrogen receptor α is developmentally regulated during osteoblast differentiation and contributes to selective responsiveness of gene expression. J. Endocrinology, 1998, 139(4): 2048-2057
[99] Korach KS, Couse JF, Curtis S W, Washburn TF, Lindzey J, Kimbro KS, Eddy EM, Migliaccio S, Snedeker SM, Lubahn DB, Schomberg DW, Smith EP. Estrogen receptor gene disruption: molecular characterization and experimental and clinical phenotypes. J. Recent Prog. Hor. Res., 1996, 51:159-188
[100] Westerlind KC, Wronski TJ, Ritman EL, Luo ZP, An KN, Bell NH, Turner RT. Estrogen regulates the rate of bone turnover but bone balance in ovariectomized rats is modulated by prevailing mechanical strain. J. Proc. Natl. Acad. Sci., 1997, 94(8):4199-4204
[101] Yasuhiro Kumei, Hitoyata Shimokawa, Hisako Katano,Eiji Hara, Hideo Akiyama, Masahiko Hirano, Chiaki Mukai, Shunji Nagaoka, Pcggy A. Whitson Microgravity induces prostag-1 anding E2 and interleukin-6 production in normal rat osteoblasts: role in bone demineralization. Journal of Biotechnology 1996, 47:313-324
[102] Horowitz MC. Cytokines and estrogen in bone: antiosteoporotic effects. Science, 1993, 260(5108): 626-627
[103] Manolagas SC, Jilka RL. Bone marro, cytokines, and bone re-modeling. N. Engl. Med. 1995, 1:332-305
[104] Vargas SJ, Napreta A, Lee SK, Kalinowski J, Kawaguchi H, Pilbeam CC, Raisz LG, Lorenzo JA. Lack of evidence for an increase in interleukin-6 expression in adult murine bone, bone marrow, and marrow stromal cell cultures after ovariecto-my. J. Bone. Miner. Res. 1996, 11(12):1926-1934
[105] Roodman GD. Interleukin-6: An osteotropic factor? J. Bone. Miner. Res. 1992, 7(5): 475-478
[106] Mundy GR. Peptides and growth regulatory factors in bone. Rheum. Dis. Clin. North Am. 1994, 20(3): 577-588
[107] Zheng SX, Vrindts Y, Lopez M, De Groote D, Zangerle PF, Collette J, Franchimont N, Geenen V, Albert A, Reginster JY. Increase in butokine production by stimulated whole bolld cells In postmenopausal osteoporosis, Maturltas. 1997, 26(1): 63-71
[108] Kozawa O, Suzuki A, Kaida T, Tokuda H, Uematsu T. Tumor necrosis factor-alpha autoregulates Interleukln-6 synthesis via activation of protein klnase C. Function of Sphin-qosine 1-phosphate and phosphatldycholine-specific phospholipase C. J Biol Chem. 1997, 272(40):25099-25104
[109] Ralston SH. Analysis of gene expression In human bone biopsies by polym erase chain reaction: evidence for enhanced cytoklne expression in postmemopausal osteoporosis. J Bone Miner Res. 1994, 9(6): 883-890
[110] Tanaka H, Barnes J, Liang CT, Effect of age on the expession of insulin Qrowth factor-1, interleukin-6 and trans forming gtrowth factor-beta mRNA in rat femurs following marrow ablation Bone. 1996, 18(5): 473-478
[111] Murray RE, Mc Guigan F, Grant SF, Reid DM, Ralston SH. Polymorphisms of the iuterleukin-6 gene are associated with bone mineral density. Bone, 1997, 21(1): 89-92
[112] Sun WH, Keller EI, Stebler BS, Ershler WB. Estrogen inhibits phorbol ester induced 1 kappa B alpha transcription and protein degradation. Biochem. Biophys Res. Commun, 1998, 244(3):691-695
[113] Kim GS, Kim CH, Choi CS, Park JY, Lee KU. Involvement of different second messengers in parathyroid hormone and inerleukin-1 induced lnterleukin-6 and interleukin-11 production in human bone marrow stromal cells. J. Bone Miner Res. 1997, 12(6): 896-902
[114] Enmark E, gustafsson JA. Newly discovered estrogen receptor, new therapeutic possibilities in post menopausal symptoms, osteoporosis, cancer of the breast and prostate. J. Lakartidningen, 1998, 95:1945-1949
[115] Wang L, Andersson S, Warner M, Gustafsson JA. Estrogen receptor (ER)beta knockout mice reveal a role for ERbeta in migration of cortical neurons in the developing brain. Proc Natl. Acad Sci U.S.A. 2003, 100 (2): 703-8.
[2] 杨彬．半胱氨酸组织蛋白酶抑制剂：一类新型的骨质疏松治疗药物．国外医学·药学分册，2000，27(2)：79-84
[3] Zhou S, Turgeman G, Harris SE, Leitman DC, Komm BS, Bodine PV, Gazit D. Estrogens activate bone morphogenetic protein-2 gene transcription in mouse mesenchymal stem cells. Mol. Endocrinol. 2003, 17(1): 56-66
[4] Lubahn DB, Moyer JS, Golding Ts, Couse JF, Korach KS, Smithies O. Alteration of reproductive function but not prenatal sexual development after insertional disruption of the mouse estrogen receptor gene J. Proc. Natl. Acad. Sci. USA 1993, 90:11162-11061
[5] Smith FP, Boyd J, Frank GR, Takahashi H, Cohen RM, Specker B, Williams TC, Lubahn DB, Korach KS. Estrogen resistance caused by a mutation in the estrogen -receptor gene in a man J N. Engl. Med. 1994, 331: 1056-1061
[6] Zofkova I, Zajickova K, Hill M. The estrogen receptor alpha gene determines serum androstenedione levels in postmenopausal women. Steroids 2002, 67(10):815-9
[7] Kobayashi N, Fujino T, Shirogane T, Furuta I, Kobamatsu Y, Yaegashi M, Sakuragi N, Fujimoto S. Estrogen receptor alpha polymorphism as a genetic marker for bone loss, vertebral fractures and susceptibility to estrogen. Maturitas 2002, 41 (3): 193-201
[8] Allcroft LC, Varanasi SS, Dimopoulos D, Francis RM, Datta HK. Mutational and Polymorphic Analysis of the Estradiol Receptor-a Gene in Men with Symptomatic Vertebral Fractures. Calcif. Tissue Int. 2002, 71(5):400-5
[9] Fujita M, Urano T, Horie K, Ikeda K, Tsukui T, Fukuoka H, Tsutsumi O, Ouchi Y, Inoue S. Estrogen activates cyclin-dependent kinases 4 and 6 through induction of cyclin D in rat primary osteoblasts. Biochem. Biophys. Res. Commun. 2002, 299(2): 222-8
[10] Ferrari SL, Ahn-Luong L, Garnero P, Humphries SE, Greenspan SL. Two promoter polymorphisms regulating interleukin-6 gene expression are associated with circulating levels of C-reactive protein and markers of bone resorption in postmenopausal women. J. Clin. Endocrinol. Metab. 2003, 88(1):255-9
[11] Taichman Rs, Reilly MJ. Emerson SG. Human osteoblasts support human hematopoietic progenitor cells in vitro bone marrow cultures. Blood 1996, 87(2): 518-24
[12] Kitamura H, Kawata H, Takahashi F, Higuchi Y, Furuichi T, Ohkawa H. Bone marrow neutrophilia and suppressed bone turnover in human interleukin-6 transgenic mice. A cellular relationship among hematopoietic cells, osteoblasts, and osteoclasts mediated by stromal cells in bone marrow. Am J Pathol 1995, 147(6): 1682-92
[13] Rude RK, Gruber HE, Wei LY, Frausto A. Magnesium deficiency: effect on bone and mineral metabolism in the mouse. Mills BG. Calcif Tissue Int 2003 Jan; 72(1): 32-41
[14] Morello KC, Wurz GT, De Gregorio MW. SERMs: current status and future trends. Crit Rev Oncol Hematol. 2002 Jul; 43(1): 63-76
[15] Bodine PV, Harris HA, Lyttle CR, Komm BS. Estrogenic effects of 7alpha-methyl-17alpha-ethynylestradiol: a newly discovered tibolone metabolite. Steroids 2002, 67(8): 681-6
[16] Hayashi M, Rho MC, Enomoto A, Fukami A, Kim YP, Kikuchi Y, Sunazuka T, Hirose T, Komiyama K, Omura S. Suppression of bone resorption by madindoline A, a novel nonpep- tide antagonist to gp130. Proc. Natl. Acad. Sci. U. S. A. 2002, 99(23): 14728-33
[17] Ishimi Y, Yoshida M, Wakimoto S, Wu J, Chiba H, Wang X, Takeda K, Miyaura C Genistein, a soybean isoflavone, affects bone marrow lymphopoiesis and prevents bone loss in castrated male mice. Bone 2002, 31(1): 180-5
[18] Torricelli P, Fini M, Giavaresi G, Giardino R. Human osteopenic bone-derived osteoblasts: essential amino acids treatment effects. Artif Cells Blood Substit. Immobil. Biotechnol. 2003, 31(1): 35-46
[19] 吴淑贤，李钢铁．补肾壮骨胶囊治疗中老年骨质疏松症临床观察报告．广州医药，1999，30(5)：71-72
[20] 张贵有．二仙坚骨汤治疗老年性脊椎骨质疏松所致腰背部疼痛65例报告．中国中医骨伤科杂志，1999，7(2)：28-29
[21] 邵敏，黄宏兴，庄洪．骨康防治骨质疏松拆方的初步研究．中国中医骨伤科杂志 2000，8(2)：7-11
[22] 王玲，仲强惟．浅谈补肾益肝健脾治疗绝经后骨质疏松症．天津中医，2001，18(1)：40-41
[23] 黄桂成．理气止痛汤治疗原发性骨质疏松腰背痛50例．镇江医学院学报，2000，10(3)：600-601
[24] 胡军．辨证治疗绝经期骨质疏松780例．中国民间疗法，2000，8(2)：40-41
[25] 陈维静，刘新梅，廉红真．中医辨证治疗原发性骨质疏松症(附109例报告)．中医药研究，1999，15(3)：29-31
[26] 陈大蓉，唐显著，郑坤渝，刘其芬，蔡汉中，李小木，张健．中药护骨合剂防治绝经后骨质疏松症的临床研究．中医杂志，1994，35(6)：359-361
[27] 危剑安，陈训华，李承军，张云如，程谟库，杨桂香，李平，华瑞成，李宝珠，王建平，康素军，陈玉英，刘丽华，石冰．肾虚骨痛胶囊对绝经期妇女骨密度的影响．中医杂志，1995，36(9)：548-550
[28] 梁克玉，魏玉玲，郭邦富，胡年宏，付刚．中药增骨汤序贯疗法治疗绝经后骨质疏松症—附120例临床观察．中医正骨，1999，11(1)：9-10
[29] 陈玉兰，牛占和，牛占山．中西医结合治疗绝经后妇女原发性骨质疏松症98例．中国骨质疏松杂志，1999，5(2)：44-46
[30] 谢雁鸣．老年OP研究近况．中医药动态，1994，(1)：11-13
[31] 程化奇，夏振信，殷新猷．更年平调冲剂治疗骨质疏松症的药理实验和临床初步研究．中国骨质疏松杂志，1999，5(1)：71-75
[32] 张玲娟，沈自尹，蔡定芳．淫羊藿对大鼠下丘脑垂体肾上腺轴抑制模型的影响．中医杂志，1996，37(10)：620-623
[33] 刘和娣，李恩，崔灵光，佟晓旭，王桂香，陈永春．补肾中药对不同年龄女性多指标及骨密度的影响．中国中西医结合杂志，1997，17(11)：671-672
[34] 沈培芝，李东煜，张戈，史万忠，徐宇，石印玉．补肾方防治地塞米松致雄性大鼠骨质疏松及其生化机制探讨．中国中西医结合杂志，1998，18(5)：290-292
[35] 刘和娣，李恩，刘昆．补肾中药对骨质疏松大鼠CaBP-D_9K基因及表达的影响．中国骨质疏松杂志，1996，2(3)：62-64
[36] 李青南，吴铁，谢华，黄莲芳，张志平．淫羊藿提取液对去睾丸大鼠骨代谢的影响．中草药，1993，24(12)：637-638
[37] 罗为民，符诗聪，杜宁，曲克服，李国衡．健脾补肾法防治绝经后骨质疏松症的计量学研究．中国中医骨伤科，1995，3(2)：1-4
[38] 陈坤，于世凤，史风芹，庞淑珍．黔岭藿对体外培养的破骨细胞作用的研究．中国骨质疏松杂志，1996，2(3)：59-61
[39] 崔家鹏，郑洪新，刘景峰，林庶茹，夏淑杰，刘树新．补肾中药对肾虚骨质疏松症大鼠红细胞膜PKCCa~(2+)-Mg~(2+)-ATP酶活性影响的实验研究．中国骨质疏松杂志，1997，3(3)：66-69
[40] 毛平，陆光伟，周志锦，杨美东，叶福媛．六味地黄丸对阴虚动物模型各个脏器组织元素含量的影响．中国中药杂志，1993：18(11)：690-692
[41] 张继东，郭福新，陈克忠．对衰老机理的认识．山东中医杂志，1992，11(6)：45-46
[42] 李芳芳，李恩，冬晓旭，陈永春，康文华．补肾、健脾和活血化瘀方药对去卵巢大鼠骨质疏松的比较性研究．中国骨质疏松杂志，1998，4(1)：5-9
[43] 林求诚，林朝晖．中医衰老指数与中医学年龄的初步分析．中西医结合杂志，1990，10(2)：90-92
[44] 何华．老年气虚血瘀初探．新中医，1991，23(7)；52-54
[45] 吕志平．中老年弦细脉甲皱微循环观察．山东中医学院学报，1994，18(2)：140-141
[46] 杨少锋．关于骨质疏松症与血瘀关系的探讨．湖南中医药导报，1996，2(4)：25-26
[47] 田代真一．“血清药理学”“血清药化学”——汉方药理学始药物中浓度测定新世界．TDM研究，1988，1(5)：54-57
[48] 田代真一．血清药理学血清药化学．现代东洋医学，1992，13(1)：113-117
[49] 季光，刘平，洪嘉和，刘成，徐列明．扶正化瘀方药物血清对原代培养纤维肝细胞胶原生成及分泌白蛋白功能的影响．中西医结合肝病杂志，1997，7(1)：25-28
[50] 何玉军，苏安英，柴锡庆，周双俊，悦随士，门金娥．用血清药理学方法观察中药复方制剂诱导人胃癌细胞凋亡的研究．中国综合临床，1998，14(1)：30-31
[51] 贺玉琢，高英杰，沈鸿，崔晓兰，郭淑英，姜廷良．给予桂枝汤的大鼠血清对HSV-1致CPE的影响．中国实验方剂学杂志，1999，5(4)：44-47
[52] 张侠，陈信义，王沛．血清药理学方法研究益肾健脾活血法对BEL-7402细胞增殖抑制作用．实用肿瘤杂志，1999，14(2)：87-89
[53] Iwama H, Amagayas S, Ogihara Y. Effect of shosaikoto, a Japanese and Chinese traditional herbal medical mixture, on the mitogenic activity of lipopolysaccharide: a new pharmacological testing method. J. Ethnopharmacol, 1987, 21(1): 45-53
[54] Umeda M, Amagayas S, Ogihara Y. Effect of certain herbal medicines on the biotrans-formation of arachidonic acid:a new pharmacological testing method using serum. J. Ethnopharmacol, 1988, 23(1): 91-98
[55] 王力倩，余上才，李仪奎，李悦．用血清药理学方法研究中药苦参、仙鹤草的抗肿瘤作用．中国中医药科技，1995，2(5)：19-21
[56] 张群豪，钟蓓，陈可冀，史大卓，毛节明，陈明哲．用血清药理学方法观察血府逐瘀浓缩丸对实验性动脉粥样硬化家兔主动脉平滑肌细胞增殖的影响．中国中西医结合杂志，1996，16(3)：156-159
[57] 崔晓兰，贺玉琢，高英杰，李晓芹，周爱香，田甲丽，郭淑英，富杭育，姜廷良．中药复方血清药理研究方法学探讨．中国实验方剂学杂志，1998，4(2)：13-15
[58] 刘成海，刘成，刘平，徐列明．扶正化瘀复方药物血清对大鼠肝贮脂细胞增殖及胶原合成的影响．中国实验方剂学杂志，1996，2(2)：16-19
[59] 刘成海，刘平，刘成，季光，徐列明．抗肝纤维化有效中药复方血清药理学方法探讨．中国实验方剂学杂志，1998，4(2)：16-19
[60] 季光，刘平，刘成，徐列明，洪嘉禾．扶正化瘀方药物血清对原代培养大鼠肝细胞增殖及胶原生成率的影响．中国实验方剂学杂志，1997，3(3)：20-23
[61] 王伟，史大卓，陈可冀，钟蓓．精制血府胶囊刺激缺氧缺糖心肌细胞RNA及蛋白质合成的研究．北京中医药大学学报，1997，20(2)：17-19
[62] 周云中，季克胜，孙小玉，丁炜，韩兴梅，洪敏．金荞麦片的血清药理学研究——对离体豚鼠回肠的影响．中成药，1999，21(1)：34-35
[63] 阴赦宏，董自波．应用血清药理学方法观察金荞麦片拮抗组胺引起的离体豚鼠回肠收缩作用．中国中医药科技，1999，6(3)：149-151
[64] 杨奎，沈映君，王一涛，曾南，张德波，史焱．含香薷、羌活胜湿汤和九味羌活丸血清对内生致热原产生的影响．中药药理与临床，1995，(4)：1-3
[65] 王力倩，金若敏，孙峥嵘，沈玉茹，陈长勋．复方仙人球对肿瘤细胞体外生长影响的实验观察．上海中医药杂志，1997，(4)：44-46
[66] 陈长勋，金若敏，贺劲松，李仪奎．用血清药理学实验方法观察附子的强心作用．中国中医药科技，1996，3(3)：12-14
[67] 李兰芳，金亚宏，原桂东，曹秀芳，姜廷良．当归补血汤抗自由基作用的药效动力学研究．中国实验方剂学杂志，1999，5(1)：24-26
[68] 杨奎，郭力，周明眉，姜远平，王一涛，蒲旭峰，李萍，陈槐卿，吴立志．中药血清药化学与中药血清药理学协同研究方法探讨．中药药理与临床，1998，14(4)：41-44
[69] 李勇，季晖，李萍，谢林．淫羊藿总黄酮对体外培养成骨细胞的影响．中国药科大学学报，2002，33(1)：48-50
[70] 高素强，傅得兴，张红梅．淫羊藿及其复方防治骨质疏松得研究进展．中国中药杂志，1999，24(4)：249-251
[71] 郑本端，吕国桥，罗自文．龟甲胶药理作用研究进展．时珍国医国药，2001，12(5)：463-465
[72] 孟大利，李铣．中药牛膝化学成分和药理活性的研究进展．中国药物化学杂志，2001，11(2)：121-122
[73] 周丕琪，沈霖，杜靖远，杨艳萍，谢晶，高兰，刘保平．补肾法对绝经后骨质疏松症患者IL-6、TNT-α的影响．中国中医骨伤科杂志，2000，8(4)：1-3
[74] 张荣华，丘和明．中医中药退行性骨质疏松症用药分析．中医药学报，1997，(4)：30-31
[75] 张荣华，陈可冀，陆大祥，马晓昌，侯励．补肾活血液延缓雄性大鼠增龄性骨质疏松的研究．中国病理生理杂志，2001，17(12)：1205-1207
[76] 冯伟，石印玉，沈培芝，徐宇，詹红生．MTT法分析中药含药血清对体外软骨细胞增殖影响的研究．上海中医药大学学报，2000，14(1)：43-45
[77] 吴健宇，李仪奎，符胜光，周芝兰．补阳还五汤保护氧自由基损伤血管内皮细胞的血清药理实验方法的建立．中药药理与临床，1999，15(1)：45-46
[78] Bergman RJ, Gazit D, Kahn AJ. Age related changes in osteogenic stem cells in mice. J. Bone Miner. Res. 1996, 1(1): 568-577
[79] 王力倩，李仪奎，符胜光，付惠娣．血清药理学方法研究探索．中药药理与临床，1997，13(3)：29-31
[80] 孟李，王宁生．含药血清的制备方法研究．中药新药与临床药理，1999，10(5)：290-292
[81] 周明眉，杨奎，姜远平，王一涛，陈槐卿，吴立志．中药血清药理学的方法学研究——采血时间的确定及时效关系研究．中药药理与临床，1999；15(1)：43-44
[82] 李仪奎．中药血清药理学实验方法的若干问题．中药新药与临床药理，1999；10(2)：95-98
[83] 周明眉，杨奎，姜远平，王一涛．中药血清药理学的方法学研究——反应体系中含药血清加入量的研究．中药药理与临床，1998，14(6)：43-44
[84] 崔晓兰，贺玉琢，高英杰，李晓芹．中药复方血清药理研究方法学探讨Ⅰ、Ⅱ．全国中药
研究学术讨论会论文摘要集．中国中西医结合学会中药专业委员会，1997：66-67
[85] 黄于贵，陈磊，贺师鹏．~3H-TdR掺入DNA技术的正确应用．中华医学杂志，1993，73(5)：308-309
[86] Owen TA, Aronow M, Shalhoub V, Barone LM, Wilming L, Tassinari MS, Kennedy MB, Pockwinse S, Lian JB, Stein GS. Progressive development of the rat osteoblast phenotype in vitro: reciprocal relationships in expression of genes associated with osteoblast proliferation and differentiation during form ation of the bone extracellular matrix. J.Cell Physiol. 1990, 143(3):420-430
[87] Pass-Even L, Gazit D, Bab I. Ontogenesis of ultrastructural features during osteogenic differentiation in diffusion chamber cultures of marrow cells. J Bone Mine. Res. 1993, 8(5):589-95
[88] J．萨姆布卢克D．W拉塞尔著．分子克隆实验指南 第三版．科学出版社出版 2002，第一版：597-701
[89] Asok K. Dasmahapatra, Barbara A.B. Wimpee, Amanda L. Trewin, Reinhold J. Hutz 2,3,7,8-Tetrachlorodibenzo-p-dioxin increases steady-state estrogen receptor- mRNA levels after CYP1A1 and CYP1B1 induction in rat granulosa cells in vitro. J. Molecular and Cellular Endocrinology 2001, 182:39-48
[90] Stoll S., Ko" nig S., Thanos S. Isolation of total-RNA from formalin-fixed rat retina. Journal of Neuroscience Methods, 2002, 120(1): 55-63
[91] Albright F. Postmenopausal osteoporosis. Trans Assoc Am Physic, 1940, 55:298-301.
[92] Jilka RL, Hangoc G, Girasole G, Passeri G, Williams DC, Abrams JS, Boyce B, Broxmeyer H, Manolagas SC. Science, 1992, 257 (5066): 88-91
[93] Tabibzadeh SS, Santhanam U, Sehgal PB, May LT. Cytokine-induced production of IFN-beta 2/IL-6 by freshly explanted human endometrial stromal cells. Modulation by estradiol-17 beta. J. Immunol, 1989, 142(9):3134-3139
[94] Chow J, Tobias JH, Colston KW, Chambers TJ. Estrogen maintains trabecular bone volume in rats not only by suppression of bone resorption but also by stimulation of bone formation. J Clin. Invest, 1992, 89(1):74-78
[95] Kuiper g Shughrue PJ, Merchenthaler I, Gustafsson JA. The estrogen receptor beta subtype: a novel mediator of estrogen action in neuroendocrine systems. Front Neuroendocrinol, 1998, 19(4):253-286
[96] Couse JF, Jonathan L, Grandien K, Gustafsson JA, Korach KS. Tissue distribution and quantitative analysis of estrogen receptor α(ERα)and estrogen receptor β(ERβ) messenger ribonucleic acid in the wild type and Er α knockout mouse. J. Endocrinology, 1997, 138(11): 4613-4612
[97] Onoe Y, Miyaura C, Ohta H, Nozawa S, Suda T. Expression of estrogen receptor β in rat bone. J. Endocrinology, 1997, 138(10): 4059-4512
[98] Bodine PV, Henderson RA, Green J, Aronow M, Owen T, Stein GS, Lian JB, Komm BS. Estrogen receptor α is developmentally regulated during osteoblast differentiation and contributes to selective responsiveness of gene expression. J. Endocrinology, 1998, 139(4): 2048-2057
[99] Korach KS, Couse JF, Curtis S W, Washburn TF, Lindzey J, Kimbro KS, Eddy EM, Migliaccio S, Snedeker SM, Lubahn DB, Schomberg DW, Smith EP. Estrogen receptor gene disruption: molecular characterization and experimental and clinical phenotypes. J. Recent Prog. Hor. Res., 1996, 51:159-188
[100] Westerlind KC, Wronski TJ, Ritman EL, Luo ZP, An KN, Bell NH, Turner RT. Estrogen regulates the rate of bone turnover but bone balance in ovariectomized rats is modulated by prevailing mechanical strain. J. Proc. Natl. Acad. Sci., 1997, 94(8):4199-4204
[101] Yasuhiro Kumei, Hitoyata Shimokawa, Hisako Katano,Eiji Hara, Hideo Akiyama, Masahiko Hirano, Chiaki Mukai, Shunji Nagaoka, Pcggy A. Whitson Microgravity induces prostag-1 anding E2 and interleukin-6 production in normal rat osteoblasts: role in bone demineralization. Journal of Biotechnology 1996, 47:313-324
[102] Horowitz MC. Cytokines and estrogen in bone: antiosteoporotic effects. Science, 1993, 260(5108): 626-627
[103] Manolagas SC, Jilka RL. Bone marro, cytokines, and bone re-modeling. N. Engl. Med. 1995, 1:332-305
[104] Vargas SJ, Napreta A, Lee SK, Kalinowski J, Kawaguchi H, Pilbeam CC, Raisz LG, Lorenzo JA. Lack of evidence for an increase in interleukin-6 expression in adult murine bone, bone marrow, and marrow stromal cell cultures after ovariecto-my. J. Bone. Miner. Res. 1996, 11(12):1926-1934
[105] Roodman GD. Interleukin-6: An osteotropic factor? J. Bone. Miner. Res. 1992, 7(5): 475-478
[106] Mundy GR. Peptides and growth regulatory factors in bone. Rheum. Dis. Clin. North Am. 1994, 20(3): 577-588
[107] Zheng SX, Vrindts Y, Lopez M, De Groote D, Zangerle PF, Collette J, Franchimont N, Geenen V, Albert A, Reginster JY. Increase in butokine production by stimulated whole bolld cells In postmenopausal osteoporosis, Maturltas. 1997, 26(1): 63-71
[108] Kozawa O, Suzuki A, Kaida T, Tokuda H, Uematsu T. Tumor necrosis factor-alpha autoregulates Interleukln-6 synthesis via activation of protein klnase C. Function of Sphin-qosine 1-phosphate and phosphatldycholine-specific phospholipase C. J Biol Chem. 1997, 272(40):25099-25104
[109] Ralston SH. Analysis of gene expression In human bone biopsies by polym erase chain reaction: evidence for enhanced cytoklne expression in postmemopausal osteoporosis. J Bone Miner Res. 1994, 9(6): 883-890
[110] Tanaka H, Barnes J, Liang CT, Effect of age on the expession of insulin Qrowth factor-1, interleukin-6 and trans forming gtrowth factor-beta mRNA in rat femurs following marrow ablation Bone. 1996, 18(5): 473-478
[111] Murray RE, Mc Guigan F, Grant SF, Reid DM, Ralston SH. Polymorphisms of the iuterleukin-6 gene are associated with bone mineral density. Bone, 1997, 21(1): 89-92
[112] Sun WH, Keller EI, Stebler BS, Ershler WB. Estrogen inhibits phorbol ester induced 1 kappa B alpha transcription and protein degradation. Biochem. Biophys Res. Commun, 1998, 244(3):691-695
[113] Kim GS, Kim CH, Choi CS, Park JY, Lee KU. Involvement of different second messengers in parathyroid hormone and inerleukin-1 induced lnterleukin-6 and interleukin-11 production in human bone marrow stromal cells. J. Bone Miner Res. 1997, 12(6): 896-902
[114] Enmark E, gustafsson JA. Newly discovered estrogen receptor, new therapeutic possibilities in post menopausal symptoms, osteoporosis, cancer of the breast and prostate. J. Lakartidningen, 1998, 95:1945-1949
[115] Wang L, Andersson S, Warner M, Gustafsson JA. Estrogen receptor (ER)beta knockout mice reveal a role for ERbeta in migration of cortical neurons in the developing brain. Proc Natl. Acad Sci U.S.A. 2003, 100 (2): 703-8.
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