柴金散结方对乳腺增生大鼠的药效作用及机制
|
摘要
目的:考察柴金散结方对大鼠乳腺增生的药效及机制,为中药新药研发提供实验依据。方法:外源性肌注雌、孕激素制备乳腺增生SD大鼠模型,造模成功后,大鼠按体质量随机分为正常组、模型组、柴金散结方(柴金)低、中、高剂量(3. 13,6. 26,12. 52 g·kg~(-1))组及乳癖消(0. 517 g·kg~(-1))组,每组9只。给药28 d,放射免疫法测定雌二醇(E_2),孕酮(P)及泌乳素(PRL),计算子宫、卵巢系数,观察乳头直径、乳腺组织病理,免疫组化法测定乳腺组织中雌激素受体-α(ER-α)表达量,实时荧光定量聚合酶链式反应(Real-time PCR)测定下丘脑、垂体促性腺激素释放激素(GnRH),促性腺激素释放激素受体(GnRH-R) mRNA表达; ICR小鼠按体质量随机分为正常组、柴金低、中、高剂量组(5. 2,10. 4,20. 8 g·kg~(-1))与罗通定组(0. 038 6 g·kg~(-1)),每组12只,给药7 d,末次给药30 min后腹腔注射0. 6%乙酸,观察15 min内扭体次数。结果:与正常组比较,模型组乳头直径增宽,血清E_2显著升高(P <0. 01),乳腺组织增生,ER-α表达增加;与模型组比较,柴金高剂量组乳头直径显著下降(P <0. 05);柴金各剂量组E_2均有下降;柴金中、高剂量组可降低乳腺增生病理积分;柴金各剂量组均能降低下丘脑中GnRH mRNA表达;柴金高剂量组小鼠扭体次数减少(P <0. 05)。结论:柴金散结方可改善乳腺增生病变情况,作用机制可能与调节下丘脑GnRH mRNA表达及降低雌激素受体表达有关。
Objective: To investigate the pharmacodynamics and mechanism of Chaijin Sanjie prescription(CJSJP) on rat mammary gland hyperplasia,in order to provide experimental basis for the research and development of new Chinese medicine. Method: SD rat model of mammary gland hyperplasia was established through exogenous intramuscular injection with estrogen and progesterone. After successful establishment of the model,the rats were randomly divided into normal group,model group,and low,medium and high-dose CJSJP groups(3. 13,6. 26,12. 52 g·kg~(-1)) and Rupixiao(0. 517 g·kg~(-1)) group,with 9 rats in each group. After 28 days of administration, estradiol(E_2), progesterone(P) and rolactin(PRL) were measured by radioimmunoassay,uterus and ovary coefficients were calculated; nipple diameter and breast histopathology were observed,estrogen receptor-α(ER-α) expression in mammary gland was measured by immunohistochemistry,and gonadotropin-releasing hormone(GnRH) and gonadotropin-releasing hormone receptor(GnRH-R) mRNA expressions in hypothalamus,pituitary were measured by Real-time PCR. ICR mice were randomly divided into normal group,low,medium and high-dose CJSJP groups(5. 2,10. 4,20. 8 g·kg~(-1)) and Luotongding group(0. 038 6 g·kg~(-1)) according to their body weight. Twelve mice in each group were given drugs for 7 days,and 0. 6% acetic acid was injected intraperitoneally for 30 minutes after the last administration. The writhing times were observed within 15 minutes. Result: Compared with the normal group,the diameter of nipple was widened,serum E_2 was significantly increased(P < 0. 01),breast tissue proliferation and ER-α expression were increased in model group. compared with model group,the diameter of nipple was significantly decreased in high dose group of CJSJP(P < 0. 05),E_2 was decreased in all dose groups of CJSJP,pathological score of breast hyperplasia was decreased in middle and high dose groups of CJSJP,GnRH mRNA in hypothalamus was decreased in all dose groups of CJSJP. The writhing times of mice in high dose group of CJSJP was decreased(P < 0. 05). Conclusion: Chaijin Sanjie prescription can improve the lesions of breast hyperplasia. The therapeutic mechanism may be related to the regulation of GnRH gene expression in hypothalamus and the decrease of estrogen receptor expression.
Objective: To investigate the pharmacodynamics and mechanism of Chaijin Sanjie prescription(CJSJP) on rat mammary gland hyperplasia,in order to provide experimental basis for the research and development of new Chinese medicine. Method: SD rat model of mammary gland hyperplasia was established through exogenous intramuscular injection with estrogen and progesterone. After successful establishment of the model,the rats were randomly divided into normal group,model group,and low,medium and high-dose CJSJP groups(3. 13,6. 26,12. 52 g·kg~(-1)) and Rupixiao(0. 517 g·kg~(-1)) group,with 9 rats in each group. After 28 days of administration, estradiol(E_2), progesterone(P) and rolactin(PRL) were measured by radioimmunoassay,uterus and ovary coefficients were calculated; nipple diameter and breast histopathology were observed,estrogen receptor-α(ER-α) expression in mammary gland was measured by immunohistochemistry,and gonadotropin-releasing hormone(GnRH) and gonadotropin-releasing hormone receptor(GnRH-R) mRNA expressions in hypothalamus,pituitary were measured by Real-time PCR. ICR mice were randomly divided into normal group,low,medium and high-dose CJSJP groups(5. 2,10. 4,20. 8 g·kg~(-1)) and Luotongding group(0. 038 6 g·kg~(-1)) according to their body weight. Twelve mice in each group were given drugs for 7 days,and 0. 6% acetic acid was injected intraperitoneally for 30 minutes after the last administration. The writhing times were observed within 15 minutes. Result: Compared with the normal group,the diameter of nipple was widened,serum E_2 was significantly increased(P < 0. 01),breast tissue proliferation and ER-α expression were increased in model group. compared with model group,the diameter of nipple was significantly decreased in high dose group of CJSJP(P < 0. 05),E_2 was decreased in all dose groups of CJSJP,pathological score of breast hyperplasia was decreased in middle and high dose groups of CJSJP,GnRH mRNA in hypothalamus was decreased in all dose groups of CJSJP. The writhing times of mice in high dose group of CJSJP was decreased(P < 0. 05). Conclusion: Chaijin Sanjie prescription can improve the lesions of breast hyperplasia. The therapeutic mechanism may be related to the regulation of GnRH gene expression in hypothalamus and the decrease of estrogen receptor expression.
引文
[1]王丽,蔡惠,叶尔买克,等.百麦外用中药制剂对小鼠乳腺增生的实验研究[J].北京联合大学学报:自然科学版,2010,24(4):73-75.
[2]顾乃强.实用中医乳房病学[M].上海:上海科学技术出版社,1993:183.
[3]张燕燕,周胜利.中医对乳腺增生病的认识及实验研究进展[J].浙江中西医结合杂志,2015,25(3):314-316.
[4]Dupont W D,Page D L.Risk factors for breast cancer in women with Proliferative breast disease[J].N Engl JMed,1985,312(3):146-151.
[5]Page D L.The woman at high risk for breast cancer importance of hyperplasia[J].Surg Clin North Am,1996,76(2):221-230.
[6]王桂玲,任连成,刘春香.乳腺增生与乳腺癌相关性研究[J].中国医药导刊,2014,16(6):972-973.
[7]张思浩,崔童星,王刚平,等.乳腺癌前期病变研究的最新进展[J].临床普外科电子杂志,2017,5(1):40-43.
[8]王肖寒.乳腺癌的发病机制与治疗[J].世界最新医学信息文摘,2017,17(63):195.
[9]Mourits M J,De Vries E G,Willemse P H,et al.Tamoxifen treatment and gynecologic side effects:a review[J].Obstet Gynecol,2001,97(5):855-866.
[10]李佳林,苏泽琦,彭莉,等.秦月好治疗乳腺增生病经验[J].中医学报,2018,33(5):778-781.
[11]苗明三,温亚娟,白明,等.乳腺增生动物模型制备规范(草案)[J].中国实验方剂学杂志,2017,23(24):17-22.
[12]段好刚,魏玉辉,李波霞,等.疏乳消块丸对大鼠乳腺增生的治疗作用及其机制[J].中国医院药学杂志,2012,32(16):1273-1276.
[13]杜力军,赵玉男.实验动物与实验动物模型[M].北京:中国医药科技出版社,2012:410.
[14]雷心雨,黄怡婷,朱翠玲,等.黄芪总皂苷配伍雷公藤多苷抗炎镇痛作用[J].中药与临床,2015,6(6):29-31.
[15]Bundred N J.Aetiological factors in benign breast disease[J].Br J Surg,1994,81(6):788-789.
[16]丁晓玲.青香乳康颗粒治疗乳腺增生病的主要药效学及机制研究[D].成都:成都中医药大学,2010.
[17]赵洋.化瘀消癖汤治疗乳腺小叶增生疗效观察[J].实用中医药杂志,2017,33(5):480-481.
[18]林旭丰,严梅娣,谢小红,等.中医周期疗法对乳腺增生伴乳痛患者内分泌激素及症状的改善观察[J].中华中医药学刊,2017,35(3):745-748.
[19]刘胜,陆德铭,唐汉钧.调摄冲任法治疗乳腺增生病的机理研究[J].辽宁中医杂志,2000,27(10):444-445.
[20]苏莹,陈兴华,黄子培,等.穴位贴敷治疗乳腺增生病的临床研究[J].针灸临床杂志,2017,33(5):4-7.
[21]黄霖,刘华,杨辉,等.健乳灵对乳腺增生大鼠GnRH及其受体mRNA表达的影响[J].中国中医急症,2006,15(9):1016-1017.
[2]顾乃强.实用中医乳房病学[M].上海:上海科学技术出版社,1993:183.
[3]张燕燕,周胜利.中医对乳腺增生病的认识及实验研究进展[J].浙江中西医结合杂志,2015,25(3):314-316.
[4]Dupont W D,Page D L.Risk factors for breast cancer in women with Proliferative breast disease[J].N Engl JMed,1985,312(3):146-151.
[5]Page D L.The woman at high risk for breast cancer importance of hyperplasia[J].Surg Clin North Am,1996,76(2):221-230.
[6]王桂玲,任连成,刘春香.乳腺增生与乳腺癌相关性研究[J].中国医药导刊,2014,16(6):972-973.
[7]张思浩,崔童星,王刚平,等.乳腺癌前期病变研究的最新进展[J].临床普外科电子杂志,2017,5(1):40-43.
[8]王肖寒.乳腺癌的发病机制与治疗[J].世界最新医学信息文摘,2017,17(63):195.
[9]Mourits M J,De Vries E G,Willemse P H,et al.Tamoxifen treatment and gynecologic side effects:a review[J].Obstet Gynecol,2001,97(5):855-866.
[10]李佳林,苏泽琦,彭莉,等.秦月好治疗乳腺增生病经验[J].中医学报,2018,33(5):778-781.
[11]苗明三,温亚娟,白明,等.乳腺增生动物模型制备规范(草案)[J].中国实验方剂学杂志,2017,23(24):17-22.
[12]段好刚,魏玉辉,李波霞,等.疏乳消块丸对大鼠乳腺增生的治疗作用及其机制[J].中国医院药学杂志,2012,32(16):1273-1276.
[13]杜力军,赵玉男.实验动物与实验动物模型[M].北京:中国医药科技出版社,2012:410.
[14]雷心雨,黄怡婷,朱翠玲,等.黄芪总皂苷配伍雷公藤多苷抗炎镇痛作用[J].中药与临床,2015,6(6):29-31.
[15]Bundred N J.Aetiological factors in benign breast disease[J].Br J Surg,1994,81(6):788-789.
[16]丁晓玲.青香乳康颗粒治疗乳腺增生病的主要药效学及机制研究[D].成都:成都中医药大学,2010.
[17]赵洋.化瘀消癖汤治疗乳腺小叶增生疗效观察[J].实用中医药杂志,2017,33(5):480-481.
[18]林旭丰,严梅娣,谢小红,等.中医周期疗法对乳腺增生伴乳痛患者内分泌激素及症状的改善观察[J].中华中医药学刊,2017,35(3):745-748.
[19]刘胜,陆德铭,唐汉钧.调摄冲任法治疗乳腺增生病的机理研究[J].辽宁中医杂志,2000,27(10):444-445.
[20]苏莹,陈兴华,黄子培,等.穴位贴敷治疗乳腺增生病的临床研究[J].针灸临床杂志,2017,33(5):4-7.
[21]黄霖,刘华,杨辉,等.健乳灵对乳腺增生大鼠GnRH及其受体mRNA表达的影响[J].中国中医急症,2006,15(9):1016-1017.