Phytosterols in hull-less pumpkin seed oil, rich in Δ7-phytosterols, ameliorate benign prostatic hyperplasia by lowing 5α-reductase and regulating balance between cell proliferation and apoptosis in rats

  • Xin-cong Kang Horticulture College, Hunan Agricultural University, Changsha, Hunan, P. R. China; Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Hunan Agricultural University, Changsha, Hunan, P. R. China; State Key Laboratory of Subhealth Intervention Technology, Changsha, Hunan, P. R. China; and Hunan Co-Innovation Center for Utilization of Botanical Functional Ingredients, Changsha, Hunan, P. R. China
  • Tian Chen Horticulture College, Hunan Agricultural University, Changsha, Hunan, P. R. China
  • Jia-li Zhou Horticulture College, Hunan Agricultural University, Changsha, Hunan, P. R. China
  • Peng-yuan Shen Horticulture College, Hunan Agricultural University, Changsha, Hunan, P. R. China
  • Si-hui Dai Horticulture College, Hunan Agricultural University, Changsha, Hunan, P. R. China
  • Chang-qing Gao Department of Laboratory Animals, Xiang Ya Hospital, Central South University, Changsha, Hunan, P. R. China
  • Jia-yin Zhang Horticulture College, Hunan Agricultural University, Changsha, Hunan, P. R. China; Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Hunan Agricultural University, Changsha, Hunan, P. R. China; State Key Laboratory of Subhealth Intervention Technology, Changsha, Hunan, P. R. China
  • Xing-yao Xiong Horticulture College, Hunan Agricultural University, Changsha, Hunan, P. R. China; Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Hunan Agricultural University, Changsha, Hunan, P. R. China; and Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, P. R. China
  • Dong-bo Liu Horticulture College, Hunan Agricultural University, Changsha, Hunan, P. R. China; Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Hunan Agricultural University, Changsha, Hunan, P. R. China; State Key Laboratory of Subhealth Intervention Technology, Changsha, Hunan, P. R. China; and Hunan Co-Innovation Center for Utilization of Botanical Functional Ingredients, Changsha, Hunan, P. R. China
DOI: https://doi.org/10.29219/fnr.v65.7537
Keywords: lower urinary tract symptoms; 5α-reductase inhibitor; MAPK pathway; apoptosis; proliferation; phytochemicals; Curcurbita pepo

Abstract

Background: Pumpkin seed oil is widely used to treat benign prostatic hyperplasia (BPH), a common disease in elder men. However, its active components and mechanism have remained to be elucidated.

Objective: The objective of the present study was to investigate the active components of pumpkin seed oil and its mechanism against BPH.

Design: Total phytosterol (TPS) was isolated from hull-less pumpkin (Cucurbita pepo L. var. Styriaca) seed oil and analyzed by gas chromatography/mass spectrometry (GC/MS). Three phytosterols were purified by preparative HPLC (high performance liquid chromatography) and confirmed by NMR (nuclear magnetic resonance). TPS (3.3 mg/kg body weight, 1 mL/day/rat) was administered intragastrically to the testosterone propionate-induced BPH rats for 4 weeks. The structure changes of prostate tissues were assessed by hematoxylin & eosin (H&E) staining. The expression of androgen receptor (AR) and steroid receptor coactivator 1 (SRC-1) was analyzed by immunohistochemistry, while that of 5α-reductase (5AR), apoptosis, or proliferation-related growth factors/proteins was detected by real-time quantitative polymerase chain reaction or western blotting.

Results: The ∆7-phytosterols in TPS reached up to 87.64%. Among them, 24β-ethylcholesta-7,22,25-trienol, 24β-ethylcholesta-7,25(27)-dien-3-ol, and ∆7-avenasterol were confirmed by NMR. TPS treatment significantly ameliorated the pathological prostate enlargement and restored histopathological alterations of prostate in BPH rats. It effectively suppressed the expressions of 5AR, AR, and coactivator SRC-1. TPS inhibited the expression of proliferation-related growth factor epidermal growth factor, whereas it increased the expressions of apoptosis-related growth factor/gene transforming growth factor-β1. The proliferation-inhibiting effect was achieved by decreasing the ERK (extracellular signal-regulated kinase) phosphorylation, while apoptosis was induced by Caspase 3 activation through JNK (c-Jun N-terminal kinase) and p38 phosphorylation.

Conclusion: TPS from hull-less pumpkin seed oil, with ∆7-phytosterols as its main ingredients, is a potential nutraceutical for BPH prevention.

Downloads

Download data is not yet available.

References


  1. Egan KB. The epidemiology of benign prostatic hyperplasia associated with lower urinary tract symptoms: prevalence and incident rates. Urol Clin North Am 2016; 43(3): 289–97. doi: 10.1016/j.ucl.2016.04.001

  2. Marberger M. Drug insight: 5alpha-reductase inhibitors for the treatment of benign prostatic hyperplasia. Nat Clin Pract Urol 2006; 3(9): 495–503. doi: 10.1038/ncpuro0577

  3. Foster CS. Pathology of benign prostatic hyperplasia. Prostate Suppl 2000; 9: 4–14. doi: 10.1002/1097-0045(2000)45:9+​<4::aid-pros3>3.0.co;2-q

  4. Untergasser G, Madersbacher S, Berger P. Benign prostatic hyperplasia: age-related tissue-remodeling. Exp Gerontol 2005; 40(3): 121–8. doi: 10.1016/j.exger.2004.12.008

  5. Carson C, 3rd, Rittmaster R. The role of dihydrotestosterone in benign prostatic hyperplasia. Urology 2003; 61(4 Suppl 1): 2–7. doi: 10.1016/s0090-4295(03)00045-1

  6. Papatsoris AG, Papavassiliou AG. Molecular ‘palpation’ of BPH: a tale of MAPK signalling? Trends Mol Med 2001; 7(7): 288–92. doi: 10.1016/s1471-4914(01)02015-9

  7. Yue J, Lopez JM. Understanding MAPK signaling pathways in apoptosis. Int J Mol Sci 2020; 21(7): 1–22. doi: 10.3390/ijms21072346

  8. Kim EH, Larson JA, Andriole GL. Management of benign prostatic hyperplasia. Annu Rev Med 2016; 67: 137–51. doi: 10.1146/annurev-med-063014-123902

  9. Allkanjari O, Vitalone A. What do we know about phytotherapy of benign prostatic hyperplasia? Life Sci 2015; 126: 42–56. doi: 10.1016/j.lfs.2015.01.023

  10. Hong H, Kim CS, Maeng S. Effects of pumpkin seed oil and saw palmetto oil in Korean men with symptomatic benign prostatic hyperplasia. Nutr Res Pract 2009; 3(4): 323–7. doi: 10.4162/nrp.2009.3.4.323

  11. Nishimura M, Ohkawara T, Sato H, Takeda H, Nishihira J. Pumpkin seed oil extracted from Cucurbita maxima improves urinary disorder in human overactive bladder. J Tradit Complement Med 2014; 4(1): 72–4. doi: 10.4103/2225-4110.124355

  12. Morakul B, Teeranachaideekul V, Buraphacheep Junyaprasert V. Niosomal delivery of pumpkin seed oil: development, characterisation, and physical stability. J Microencapsul 2019; 36(2): 120–9. doi: 10.1080/02652048.2019.1607597

  13. Stevenson DG, Eller FJ, Wang L, Jane JL, Wang T, Inglett GE. Oil and tocopherol content and composition of pumpkin seed oil in 12 cultivars. J Agric Food Chem 2007; 55(10): 4005–13. doi: 10.1021/jf0706979

  14. Montesano D, Blasi F, Simonetti MS, Santini A, Cossignani L. Chemical and nutritional characterization of seed oil from Cucurbita maxima L. (var. Berrettina) pumpkin. Foods 2018; 7(3): 1–14. doi: 10.3390/foods7030030

  15. Klippel KF, Hiltl DM, Schipp B. A multicentric, placebo-controlled, double-blind clinical trial of beta-sitosterol (phytosterol) for the treatment of benign prostatic hyperplasia. German BPH-Phyto Study group. Br J Urol 1997; 80(3): 427–32. doi: 10.1046/j.1464-410x.1997.t01-1-00362

  16. Wilt TJ, MacDonald R, Ishani A. beta-sitosterol for the treatment of benign prostatic hyperplasia: a systematic review. BJU Int 1999; 83(9): 976–83. doi: 10.1046/j.1464-410x.1999.00026.x

  17. Wang R, Kobayashi Y, Lin Y, Rauwald HW, Fang L, Qiao H, et al. A phytosterol enriched refined extract of Brassica campestris L. pollen significantly improves benign prostatic hyperplasia (BPH) in a rat model as compared to the classical TCM pollen preparation Qianlie Kang Pule’an Tablets. Phytomedicine 2015; 22(1): 145–52. doi: 10.1016/j.phymed.2014.10.001

  18. Tsai YS, Tong YC, Cheng JT, Lee CH, Yang FS, Lee HY. Pumpkin seed oil and phytosterol-F can block testosterone/prazosin-induced prostate growth in rats. Urol Int 2006; 77(3): 269–74. doi: 10.1159/000094821

  19. Schmittgen TD, Livak KJ. Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc 2008; 3(6): 1101–8. doi: 10.1038/nprot.2008.73

  20. Vipin K. Garg WRN. Occurrence of Δ5-sterols in plants producing predominantly Δ7-sterols: studies on the sterol compositions of six cucurbitaceae seeds. Phytochemistry 1986; 25(11): 2591–7. doi: 10.1016/s0031-9422(00)84516-1

  21. Zhang X, Cambrai A, Miesch M, Roussi S, Raul F, Aoude-Werner D, et al. Separation of Delta5- and Delta7-phytosterols by adsorption chromatography and semipreparative reversed phase high-performance liquid chromatography for quantitative analysis of phytosterols in foods. J Agric Food Chem 2006; 54(4): 1196–202. doi: 10.1021/jf052761x

  22. Berges RR, Windeler J, Trampisch HJ, Senge T. Randomised, placebo-controlled, double-blind clinical trial of beta-sitosterol in patients with benign prostatic hyperplasia. Beta-sitosterol Study Group. Lancet 1995; 345(8964): 1529–32. doi: 10.1016/s0140-6736(95)91085-9

  23. Awad AB, Fink CS, Williams H, Kim U. In vitro and in vivo (SCID mice) effects of phytosterols on the growth and dissemination of human prostate cancer PC-3 cells. Eur J Cancer Prev 2001; 10(6): 507–13. doi: 10.1097/00008469-200112000-00005

  24. Cabeza M, Bratoeff E, Heuze I, Ramirez E, Sanchez M, Flores E. Effect of beta-sitosterol as inhibitor of 5 alpha-reductase in hamster prostate. Proc West Pharmacol Soc 2003; 46: 153–5. PMID: 14699915.

  25. Roehrborn CG. Pathology of benign prostatic hyperplasia. Int J Impot Res 2008; 20 Suppl 3: S11–8. doi: 10.1038/ijir.2008.55

  26. Wang X, Lin WJ, Izumi K, Jiang Q, Lai KP, Xu D, et al. Increased infiltrated macrophages in benign prostatic hyperplasia (BPH): role of stromal androgen receptor in macrophage-induced prostate stromal cell proliferation. J Biol Chem 2012; 287(22): 18376–85. doi: 10.1074/jbc.M112.355164

  27. Kaplan SA, Roehrborn CG, McConnell JD, Meehan AG, Surynawanshi S, Lee JY, et al. Long-term treatment with finasteride results in a clinically significant reduction in total prostate volume compared to placebo over the full range of baseline prostate sizes in men enrolled in the MTOPS trial. J Urol 2008; 180(3): 1030–2; discussion 2–3. doi: 10.1016/j.juro.2008.05.004

  28. Youn DH, Park J, Kim HL, Jung Y, Kang J, Lim S, et al. Corrigendum: berberine improves benign prostatic hyperplasia via suppression of 5 alpha reductase and extracellular signal-regulated kinase in vivo and in vitro. Front Pharmacol 2019; 10: 541. doi: 10.3389/fphar.2019.00541

  29. Minutoli L, Rinaldi M, Marini H, Irrera N, Crea G, Lorenzini C, et al. Apoptotic pathways linked to endocrine system as potential therapeutic targets for benign prostatic hyperplasia. Int J Mol Sci 2016; 17(8): 1–15. doi: 10.3390/ijms17081311

  30. Kyprianou N, Tu H, Jacobs SC. Apoptotic versus proliferative activities in human benign prostatic hyperplasia. Hum Pathol 1996; 27(7): 668–75. doi: 10.1016/s0046-8177(96)90396-2

  31. Meloche S, Pouyssegur J. The ERK1/2 mitogen-activated protein kinase pathway as a master regulator of the G1- to S-phase transition. Oncogene 2007; 26(22): 3227–39. doi: 10.1038/sj.onc.1210414

  32. MacCorkle RA, Tan TH. Mitogen-activated protein kinases in cell-cycle control. Cell Biochem Biophys 2005; 43(3): 451–61. doi: 10.1385/CBB:43:3:451

  33. Tsai SC, Huang WW, Huang WC, Lu CC, Chiang JH, Peng SF, et al. ERK-modulated intrinsic signaling and G(2)/M phase arrest contribute to the induction of apoptotic death by allyl isothiocyanate in MDA-MB-468 human breast adenocarcinoma cells. Int J Oncol 2012; 41(6): 2065–72. doi: 10.3892/ijo.2012.1640

  34. Youn DH, Park J, Kim HL, Jung Y, Kang J, Jeong MY, et al. Chrysophanic acid reduces testosterone-induced benign prostatic hyperplasia in rats by suppressing 5alpha-reductase and extracellular signal-regulated kinase. Oncotarget 2017; 8(6): 9500–12. doi: 10.18632/oncotarget.13430

  35. Wells A, Gupta K, Chang P, Swindle S, Glading A, Shiraha H. Epidermal growth factor receptor-mediated motility in fibroblasts. Microsc Res Tech 1998; 43(5): 395–411. doi: 10.1002/(SICI)1097-0029(19981201)43:5<395::AID-JEMT6>3.0.CO;2-T

  36. Davis RJ. Signal transduction by the JNK group of MAP kinases. Cell 2000; 103(2): 239–52. doi: 10.1016/s0092-8674(00)00116-1

  37. Ropiquet F, Giri D, Lamb DJ, Ittmann M. FGF7 and FGF2 are increased in benign prostatic hyperplasia and are associated with increased proliferation. J Urol 1999; 162(2): 595–9. doi: 10.1016/S0022-5347(05)68632-6

  38. Lucia MS, Lambert JR. Growth factors in benign prostatic hyperplasia: basic science implications. Curr Urol Rep 2008; 9(4): 272–8. doi: 10.1007/s11934-008-0048-6

  39. Poncet N, Guillaume J, Mouchiroud G. Epidermal growth factor receptor transactivation is implicated in IL-6-induced proliferation and ERK1/2 activation in non-transformed prostate epithelial cells. Cell Signal 2011; 23(3): 572–8. doi: 10.1016/j.cellsig.2010.11.009

  40. Sreenivasulu K, Nandeesha H, Dorairajan LN, Rajappa M, Vinayagam V, Cherupanakkal C. Gene expression of insulin receptor, insulin-like growth factor increases and insulin-like growth factor-binding protein-3 reduces with increase in prostate size in benign prostatic hyperplasia. Aging Male 2018; 21(2): 138–44. doi: 10.1080/13685538.2017.1401994

  41. Yang Y, Chisholm GD, Habib FK. Epidermal growth factor and transforming growth factor alpha concentrations in BPH and cancer of the prostate: their relationships with tissue androgen levels. Br J Cancer 1993; 67(1): 152–5. doi: 10.1038/bjc.1993.26

  42. Hennenberg M, Schreiber A, Ciotkowska A, Rutz B, Waidelich R, Strittmatter F, et al. Cooperative effects of EGF, FGF, and TGF-beta1 in prostate stromal cells are different from responses to single growth factors. Life Sci 2015; 123: 18–24. doi: 10.1016/j.lfs.2014.12.006

  43. Wei H, Wu G, Shi D, Song S, Zhang X, Lei Y, et al. Total flavan glycoside from Abacopteris penangiana rhizomes and its acid hydrolysate: characterisation and anti-benign prostatic hyperplasia potential. Food Chem 2012; 134(4): 1959–66. doi: 10.1016/j.foodchem.2012.03.128

  44. Hayward SW, Wang Y, Cao M, Hom YK, Zhang B, Grossfeld GD, et al. Malignant transformation in a nontumorigenic human prostatic epithelial cell line. Cancer Res 2001; 61(22): 8135–42. PMID: 11719442.

  45. Ilio KY, Sensibar JA, Lee C. Effect of TGF-beta 1, TGF-alpha, and EGF on cell proliferation and cell death in rat ventral prostatic epithelial cells in culture. J Androl 1995; 16(6): 482–90. doi: 10.1002/j.1939-4640.1995.tb00568.x

  46. Martikainen P, Kyprianou N, Isaacs JT. Effect of transforming growth factor-beta 1 on proliferation and death of rat prostatic cells. Endocrinology 1990; 127(6): 2963–8. doi: 10.1210/endo-127-6-2963

  47. Derynck R, Zhang YE. Smad-dependent and Smad-independent pathways in TGF-beta family signalling. Nature 2003; 425(6958): 577–84. doi: 10.1038/nature02006

Published
2021-12-02
How to Cite
Kang X.- cong, Chen T., Zhou J.- li, Shen P.- yuan, Dai S.- hui, Gao C.- qing, Zhang J.- yin, Xiong X.- yao, & Liu D.- bo. (2021). Phytosterols in hull-less pumpkin seed oil, rich in Δ7-phytosterols, ameliorate benign prostatic hyperplasia by lowing 5α-reductase and regulating balance between cell proliferation and apoptosis in rats. Food & Nutrition Research, 65. https://doi.org/10.29219/fnr.v65.7537
Issue
Vol 65 (2021)
Section
Original Articles
Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Authors retain copyright of their work, with first publication rights granted to SNF Swedish Nutrition Foundation. Read the full Copyright- and Licensing Statement.

 

 

Crossref
Scopus
Google Scholar
Europe PMC