Efficacy of Lubeikangru formulation in mice with hyperplasia of the mammary glands induced by estrogen and progesterone
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摘要
OBJECTIVE: To evaluate the protective effects of Lubeikangru formula(LF) on hyperplasia of the mammary glands(HMG) induced by estrogen and progesterone in mice.METHODS: Female mice were divided randomly into five groups: normal, model, tamoxifen(3 mg/kg),Rupixiao(900 mg/kg) and LF(900 mg/kg). All mice except those in the normal group were treated sequentially with estradiol and progesterone to induce HMG. From the tenth day of induction, mice in normal and model groups received distilled water and mice in the other groups were given the corresponding drugs by gavage, once a day, for 30 d.At the end of treatment, the mammary glands, ovaries, hypothalamus, and serum was collected for whole-mount and hematoxylin and eosin(HE)staining, enzyme-linked immunosorbent assays(ELISAs), or western blotting.RESULTS: Whole-mount and HE staining of mammary glands showed that LF rescued(at least in part) the hyperplasic morphology of the mammary glands, and the number of branch points decreased after LF treatment(P < 0.05). ELISAs revealed that levels of estrogen and progesterone were decreased following LF treatment, whereas levels of gonadotropin-releasing hormone, follicle-stimulating hormone, and luteinizing hormone were increased in serum and tissues. Western blotting confirmed that LF treatment led to a reduction in expression of phosphorylated(p)-Erk, p-p38 and p-c-Jun N-terminal kinase. LF was also confirmed to be safe by acute-toxicity tests.CONCLUSION: LF can protect the mammary glands of mice from estrogen-and progesterone-induced hyperplasia by adjusting hormone levels and regulating the mitogen-activated protein kinase pathway.
OBJECTIVE: To evaluate the protective effects of Lubeikangru formula(LF) on hyperplasia of the mammary glands(HMG) induced by estrogen and progesterone in mice.METHODS: Female mice were divided randomly into five groups: normal, model, tamoxifen(3 mg/kg),Rupixiao(900 mg/kg) and LF(900 mg/kg). All mice except those in the normal group were treated sequentially with estradiol and progesterone to induce HMG. From the tenth day of induction, mice in normal and model groups received distilled water and mice in the other groups were given the corresponding drugs by gavage, once a day, for 30 d.At the end of treatment, the mammary glands, ovaries, hypothalamus, and serum was collected for whole-mount and hematoxylin and eosin(HE)staining, enzyme-linked immunosorbent assays(ELISAs), or western blotting.RESULTS: Whole-mount and HE staining of mammary glands showed that LF rescued(at least in part) the hyperplasic morphology of the mammary glands, and the number of branch points decreased after LF treatment(P < 0.05). ELISAs revealed that levels of estrogen and progesterone were decreased following LF treatment, whereas levels of gonadotropin-releasing hormone, follicle-stimulating hormone, and luteinizing hormone were increased in serum and tissues. Western blotting confirmed that LF treatment led to a reduction in expression of phosphorylated(p)-Erk, p-p38 and p-c-Jun N-terminal kinase. LF was also confirmed to be safe by acute-toxicity tests.CONCLUSION: LF can protect the mammary glands of mice from estrogen-and progesterone-induced hyperplasia by adjusting hormone levels and regulating the mitogen-activated protein kinase pathway.
OBJECTIVE: To evaluate the protective effects of Lubeikangru formula(LF) on hyperplasia of the mammary glands(HMG) induced by estrogen and progesterone in mice.METHODS: Female mice were divided randomly into five groups: normal, model, tamoxifen(3 mg/kg),Rupixiao(900 mg/kg) and LF(900 mg/kg). All mice except those in the normal group were treated sequentially with estradiol and progesterone to induce HMG. From the tenth day of induction, mice in normal and model groups received distilled water and mice in the other groups were given the corresponding drugs by gavage, once a day, for 30 d.At the end of treatment, the mammary glands, ovaries, hypothalamus, and serum was collected for whole-mount and hematoxylin and eosin(HE)staining, enzyme-linked immunosorbent assays(ELISAs), or western blotting.RESULTS: Whole-mount and HE staining of mammary glands showed that LF rescued(at least in part) the hyperplasic morphology of the mammary glands, and the number of branch points decreased after LF treatment(P < 0.05). ELISAs revealed that levels of estrogen and progesterone were decreased following LF treatment, whereas levels of gonadotropin-releasing hormone, follicle-stimulating hormone, and luteinizing hormone were increased in serum and tissues. Western blotting confirmed that LF treatment led to a reduction in expression of phosphorylated(p)-Erk, p-p38 and p-c-Jun N-terminal kinase. LF was also confirmed to be safe by acute-toxicity tests.CONCLUSION: LF can protect the mammary glands of mice from estrogen-and progesterone-induced hyperplasia by adjusting hormone levels and regulating the mitogen-activated protein kinase pathway.
引文
1 Onstad M,Stuckey A.Benign breast disorders.Obstet Gynecol Clin North Am 2013;40(3):459-473.
2 Schillace RV,Skinner AM,Pommier RF,et al.Estrogen receptor,progesterone receptor,interleukin-6 and interleukin-8 are variable in breast cancer and benign stem/progenitor cell populations.BMC Cancer 2014;14:733.
3 Visscher DW,Frank RD,Carter JM,et al.Breast cancer risk and progressive histology in serial benign biopsies.JNatl Cancer Inst 2017;109(10):djx035.
4 Verkooijen HM,Bouchardy C,Vinh-Hung V,et al.The incidence of breast cancer and changes in the use of hormone replacement therapy:a review of the evidence.Maturitas 2009;64(2):80-85.
5 Zhang G,Li D,Guo H,et al.Modulation of expression of p16 and her2 in rat breast tissues of mammary hyperplasia model by external use of rupifang extract.J Tradit Chin Med 2012;32(4):651-656.
6 Qian LQ,Pei XH,Xu ZY,et al.Clinical observation on treatment of hyperplasia of mammary gland by Lirukang Granule.Chin J Integr Med 2007;13(2):120-124.
7 Liu X,Shi Y.Treating breast hyperplasia in TCM.Zhong Yi Lin Chuang Yan Jiu 2015;7(7):147-148.
8 Wu F,Li H,Jin L,et al.Deer antler base as a Traditional Chinese Medicine:a review of its traditional uses,chemistry and pharmacology.J Ethnopharmacol 2013;145(2):403-415.
9 Sun L,Guo DH,Liu F,et al.A mouse model of mammary hyperplasia induced by oral hormone administration.Afr J Tradit Complement Altern Med 2017;14(4):247-252.
10 Huebner RJ,Neumann NM,Ewald AJ.Mammary epithelial tubes elongate through MAPK-dependent coordination of cell migration.Development 2016;143(6):983-993.
11 Fata JE,Mori H,Ewald AJ,et al.The MAPK(ERK-1,2)pathway integrates distinct and antagonistic signals from TGFalpha and FGF7 in morphogenesis of mouse mammary epithelium.Dev Biol 2007;306(1):193-207.
12 Ferin M.The Hypothalamic-hypophyseal-ovarian axis and the menstrual cycle.Global Library of Women's Medicine,2008-11,cited 2017-03-01.Available from URL:http://www.glowm.com/resources/glowm/cd/pages/v5/v5c006.html?SESSID=i2lc0la2uamk1o946gavmolc44.
13 Arendt LM,Kuperwasser C.Form and function:how estrogen and progesterone regulate the mammary epithelial hierarchy.J Mammary Gland Biol Neoplasia 2015;20(1-2):9-25.
14 Samoli E,Trichopoulos D,Lagiou A,et al.The hormonal profile of benign breast disease.Br J Cancer 2013;108(1):199-204.
15 Pesiri V,Totta P,Marino M,et al.Ubiquitin-activating enzyme is necessary for 17β-estradiol-induced breast cancer cell proliferation and migration.IUBMB Life 2014;66(8):578-585.
16 Salazar M,Lerma-Ortiz A,Hooks GM,et al.Progestin-mediated activation of MAPK and AKT in nuclear progesterone receptor negative breast epithelial cells:The role of membrane progesterone receptors.Gene 2016;591(1):6-13.
17 O'Leary B,Finn RS,Turner NC.Treating cancer with selective CDK4/6 inhibitors.Nat Rev Clin Oncol 2016;13(7):417-430.
2 Schillace RV,Skinner AM,Pommier RF,et al.Estrogen receptor,progesterone receptor,interleukin-6 and interleukin-8 are variable in breast cancer and benign stem/progenitor cell populations.BMC Cancer 2014;14:733.
3 Visscher DW,Frank RD,Carter JM,et al.Breast cancer risk and progressive histology in serial benign biopsies.JNatl Cancer Inst 2017;109(10):djx035.
4 Verkooijen HM,Bouchardy C,Vinh-Hung V,et al.The incidence of breast cancer and changes in the use of hormone replacement therapy:a review of the evidence.Maturitas 2009;64(2):80-85.
5 Zhang G,Li D,Guo H,et al.Modulation of expression of p16 and her2 in rat breast tissues of mammary hyperplasia model by external use of rupifang extract.J Tradit Chin Med 2012;32(4):651-656.
6 Qian LQ,Pei XH,Xu ZY,et al.Clinical observation on treatment of hyperplasia of mammary gland by Lirukang Granule.Chin J Integr Med 2007;13(2):120-124.
7 Liu X,Shi Y.Treating breast hyperplasia in TCM.Zhong Yi Lin Chuang Yan Jiu 2015;7(7):147-148.
8 Wu F,Li H,Jin L,et al.Deer antler base as a Traditional Chinese Medicine:a review of its traditional uses,chemistry and pharmacology.J Ethnopharmacol 2013;145(2):403-415.
9 Sun L,Guo DH,Liu F,et al.A mouse model of mammary hyperplasia induced by oral hormone administration.Afr J Tradit Complement Altern Med 2017;14(4):247-252.
10 Huebner RJ,Neumann NM,Ewald AJ.Mammary epithelial tubes elongate through MAPK-dependent coordination of cell migration.Development 2016;143(6):983-993.
11 Fata JE,Mori H,Ewald AJ,et al.The MAPK(ERK-1,2)pathway integrates distinct and antagonistic signals from TGFalpha and FGF7 in morphogenesis of mouse mammary epithelium.Dev Biol 2007;306(1):193-207.
12 Ferin M.The Hypothalamic-hypophyseal-ovarian axis and the menstrual cycle.Global Library of Women's Medicine,2008-11,cited 2017-03-01.Available from URL:http://www.glowm.com/resources/glowm/cd/pages/v5/v5c006.html?SESSID=i2lc0la2uamk1o946gavmolc44.
13 Arendt LM,Kuperwasser C.Form and function:how estrogen and progesterone regulate the mammary epithelial hierarchy.J Mammary Gland Biol Neoplasia 2015;20(1-2):9-25.
14 Samoli E,Trichopoulos D,Lagiou A,et al.The hormonal profile of benign breast disease.Br J Cancer 2013;108(1):199-204.
15 Pesiri V,Totta P,Marino M,et al.Ubiquitin-activating enzyme is necessary for 17β-estradiol-induced breast cancer cell proliferation and migration.IUBMB Life 2014;66(8):578-585.
16 Salazar M,Lerma-Ortiz A,Hooks GM,et al.Progestin-mediated activation of MAPK and AKT in nuclear progesterone receptor negative breast epithelial cells:The role of membrane progesterone receptors.Gene 2016;591(1):6-13.
17 O'Leary B,Finn RS,Turner NC.Treating cancer with selective CDK4/6 inhibitors.Nat Rev Clin Oncol 2016;13(7):417-430.