Cissampelos capensis Rhizome Extract Induces Intracellular ROS Production, Capacitation and DNA Fragmentation in Human Spermatozoa
More than 3000 plants of notable phyto-therapeutic
value grow in South Africa; these include Cissampelos capensis,
commonly known in Afrikaans as dawidjie or dawidjiewortel. C.
capensis is the most significant and popular medicinal plant used by
the Khoisan as well as other rural groups in the Western region of
South Africa. Its rhizomes are traditionally used to treat male fertility
problems. Yet, no studies have investigated the effects of this plant or
its extracts on human spermatozoa. Therefore, this study aimed at
investigating the effects of C. capensis rhizome extract (CRE)
fractions on ejaculated human spermatozoa in vitro. Spermatozoa
from a total of 77 semen samples were washed with human tubular
fluid medium supplemented with bovine serum albumin (HTF-BSA)
and incubated for 2 hours with 20 μg/ml progesterone (P4) followed
by incubation with different concentrations (0, 0.05, 0.5, 5, 50, 200
μg/ml) of fractionated CRE (F1=0% MeOH, F2=30% MeOH,
F3=60% MeOH and F4=100% MeOH) for 1.5 hours at 37°C. A
sample without addition of CRE fractions served as control. Samples
were analyzed for sperm motility, reactive oxygen species (ROS),
DNA-fragmentation, acrosome reaction and capacitation. Results
showed that F1 resulted in significantly higher values for ROS,
capacitation and hyper-activation compared to F2, F3, and F4 with
P4-stimulated samples generally having higher values. No significant
effect was found for the other parameters. In conclusion, alkaloids
present in F1 of CRE appear to have triggered sperm intrinsic ROS
production leading to sperm capacitation and acrosome reaction
induced by P4.
[1] N. E. Skakkebaek, N. Jorgensen, K. M. Main, E. Rajpert-De Meyts, H.
Leffers, A. Andersson, A. Juul, E. Carlsen, G. K. Mortensen, T. K.
Jensen, J. Toppari, “Human fecundity declining,”Int J Androl, vol. 29,
2006, pp.2-11.
[2] J. R. Sherins, “How is male fertility identified? In: Robaire B., Pryor J.
L., Trailer J.M. (eds.), Handbook of Andrology. American Society of
Andrology. Allen Press 1995, PP. 48-51.
[3] E. O. Laumann, A. Park and R. C. Rosen, “Sexual Dysfunction in
United States, Prevalence & Predictors,” JAMA, no. 6, 1999, pp. 281.
[4] H. G. Tempest, S. T. Homa, E. J. Routledge, A. Garner, X. P. and D. K.
Zhai, “Plants used in Chinese medicine for the treatment of male
infertility possess antioxidant and antioestrogenic activity,”
SystBiolReprod Med,VOL. 54, 2008, PP. 185-195.
[5] J. B. Calixto, “Efficacy, safety, quality control, marketing and regulatory
guidelines for herbal medicines (phytotherapeutic agents),”. Braz J Med
Biol Res, vol. 33, 2000, pp. 179-189.
[6] R. Yuan, and Y. Lin, “Traditional Chinese medicine: an approach to
scientific proof and clinical validation,” Pharmcol Therapeut, vol. 86,
2000, pp. 191-198.
[7] C. K. Christopher and M. T. Ho·Hui, “Rise of Herbal and Traditional
Medicine in Erectile Dysfunction management,” CurrUrol Rep, vol. 12,
2011, pp. 470-478.
[8] A. Adimoelja, “Phytochemicals and the breakthrough of traditional
herbs in the management of sexual dysfunctions,” Int J Androl, vol. 2,
2000, PP. 82-84.
[9] E. Nocerino, M. Amato and A. A. Izzo, “The aphrodisiac and
adaptogenic properties of ginseng,”Fitoterapia, vol. 71, pp. S1–S5.
[10] N. Erasmus, M. C. Solomom, K. A. Fortuin, and R. R. Henkel, “Effect
of Eurycomalongifolia Jack (Tongkatali) extract on human spermatozoa
in vitro,” Andrologia, vol. 44, 2012, pp. 308-314.
[11] C. Gonzales, J. Leiva-Revilla, J. Rubio, M. Gasco and G. F. Gonzales,
“Effect of red maca (Lepidium meyenii) on prostate zinc levels in rats
with testosterone-induced prostatic hyperplasia,” Andrologia, vol. 44,
2012, pp. 362-369.
[12] P. F. Moundipa, S. Ngouela, P. Kamtchouing, E. Tsamo, F. M.
Tchouanguep and S. Carreau,“Effects of extracts from Hibiscus
macranthus and Basella alba mixture on testosterone production in vitro
in adult rat testes slices,” Asian J Androl, vol. 8, 2006, pp. 111-114.
[13] F. Lampiao, D. Krom and S. S. du Plessis, “The in vitro effects of
Mondia whitei on human sperm motility parameters,” Phytother Res vol.
22, 2008, pp.1272-1273.
[14] J. J. Meyer Marian, C. N. Rakuambo and A. A. Hussen, “Novel
xanthones from Securidacalongependunculata with activity against
erectile dysfunction,” JEthnopharmacol, vol. 119, 2008, pp. 559-603.
[15] E. E. van Wyk, B. van Oudtshoorn and N. Gericke,“Medicinal Plants of
South Africa,” 2nd edition; Briza Publications 2009, Pretoria, South
Africa.
[16] B. E. van Wyk and N. Gericke, “People’s Plants. A Guide to the Useful
Plants of Southern Africa,” Briza 2000, Pretoria, South Africa.
[17] B. E. Van Wyk, F. R. van Heerden and B. van Oudtshoorn, “Poisonous
Plants of South Africa,” Briza Publications 2002, Pretoria, South Africa.
[18] H. De Wet and B. E. van Wyk, “An ethno-botanical survey of southern
African Menispermaceae,”S Afr J Bot, vol. 74, 2008, pp. 2-9.
[19] M. J. Watt and M. G. Breyer-Brandwijk, “The medicinal and poisonous
plants of southern and eastern Africa,” 2nd edition. Livingston, London.
[20] C. A. Smith “Common names of South African plants,” Memoirs of the
botanical survey of South Africa, no. 35, 1966, department of agriculture
and technical services.
[21] E. Von Koenen, “Medicinal, Poisonous, and Edible Plants of Namibia,”
Klaus Hess Publishers/Verlag, Windhoek 2001, Namibia.
[22] S. M. Shalaweh, N. Erasmus, F. Weitz and R. R. Henkel, “Effect of
Cissampelos capensis rhizome extract on human spermatozoa in vitro,”
Andrologia, doi: 10.1111/and.12264,2014, pp. 1-10.
[23] R. Henkel, E. Kierspel, M. Hajimohammad, T. Stalf, C. Hoogendijk, C.
Mehnert, R. Menkveld, W. Schill and T. F. Kruger, “DNA
fragmentation of spermatozoa and assisted reproduction technology,”
Reprod Biomed Online, vol. 7,2013, pp. 477-484.
[24] R. Henkel, M. Hajimohammad, T. Stalf, C. Hoogendijk, C. Mehnert, R.
Menkveld, H. Gips, W. B. Schill and T. F. Kruger, “Influence of
deoxyribonucleic acid damage on fertilization and pregnancy,”
FertilSteril, vol. 81, 2004,pp. 965-672.
[25] W. Gorczyca, F. Traganos, H. Jesionowska and Z. Daryznkiewcz,
“Presence of strand breaks and increased sensitivity of DNA in situ to
denaturation in abnormal human sperm cells: analogy to apoptosis in
somatic cells,”Exp Cell Res, vol. 207, 2003, pp. 202-205.
[26] C. M. Green, S. M. Cockle, P. F. Watson and L. R. Fraser, “Fertilization
promoting peptide, a tripeptide similar to thyrotrophin-releasing
hormone, stimulates the capacitation and fertilizing ability of human
spermatozoa in vitro,” Hum Reprod, vol. 11, 1996, pp. 830-836.
[27] WHO, “Global Atlas of Traditional, Complementary and Alternative
Medicine,” vol. 1 and 2, World Health Organization 2005, Geneva.
[28] D. S. Fabricant and N. R. Farnsworth, “The value of plants used in
traditional medicine for drug discovery,” Environmental health
perspectives, vol. 109, 2001, pp. 69.
[29] H. Azaizeh, S. Fulder, K. Khalil and O. Said, “Ethnobotanical
knowledge of local Arab practitioners in the Middle Eastern region,”
Fitoterapia,vol. 74, 2003, pp. 98-108.
[30] R. A. Halberstein, “Medicinal plants: historical and cross-cultural usage
patterns,” Annals of epidemiology, vol. 15, 2005, pp. 686-699.
[31] P. J. Houghton, “The role of plants in traditional medicine and current
therapy,” J & C M, vol. 1, 1995, pp. 131-143.
[32] R. Yuan and Y. Lin, “Traditional Chinese medicine: an approach to
scientific proof and clinical validation,” Pharmacology & therapeutics,
vol. 86, 2000,pp. 191-198.
[33] M. Solomon and R. R. Henkel, “In vivo effects of Eurycomalongifolia
Jack (Tongkat Ali) extract on reproductive functions in the
rat,”Andrologia, vol. 46 ,2014, pp. 339-348.
[34] H. De Wet, F. R. van Heerden and B. E. van Wyk, “Alkaloidal variation
in Cissampeloscapensis (Menispermaceae),” Molecules, vol. 16, 2011,
pp. 3001-3009.
[35] E. DeLamirande, P. Leclerc and C. Gagnon, “Capacitation as a
regulatory event that primes spermatozoa for the acrosome reaction and
fertilization,” Mol. Hum. Reprod, vol. 3, 1997,pp. 175–194.
[36] P. Olds-Clarke, “Unresolved issues in mammalian fertilization,”Int. Rev.
Cyt, vol, 232, 2003, pp. 129–184.
[37] C. De Jonge, “Biological basis for human capacitation,” Hum. Reprod,
vol. 11, 2005, pp. 205–214.
[38] C. O’Flaherty, E. de Lamirande and C. Gagnon, “Positive role of
reactive oxygen species in mammalian sperm capacitation triggering and
modulation of phosphorylation events, Free Radic,”Biol. Med, vol. 41,
2006, pp. 528–540.
[39] E. DeLamirande and C. O'Flaherty, “Sperm activation: role of reactive
oxygen species and kinases,” Biochim Biophys Acta, vol. 1, 2008, pp.
106-115.
[40] B. Zhao, Y. Chen, X. Sun, M. Zhou, J. Ding, J. J. Zhan andL. J. Guo,
“Phenolic alkaloids from Menispermum dauricum rhizome protect
against brain ischemia injury via regulation of GLT-1, EAAC1 and ROS
generation,” Molecules, vol. 17, 2012, pp. 2725-2737.
[41] W. H. Chiu, S. J. Luo, C.L Chen, J. H. Cheng, C. Y. Hsieh, C. Y. Wang,
Huang, W. C. Su and C. F. Lin, “Vinca alkaloids cause aberrant ROSmediated
JNK activation, Mcl-1 downregulation, DNA damage,
mitochondrial dysfunction, and apoptosis in lung adenocarcinoma cells,”
Biochem Pharmacol, vol. 83, 2012,pp. 1159-1171.
[42] H. A. Jung, B. S. Min, T. Yokozawa, J. H. Lee, Y. S. Kim and J. S.
Choi, “Anti-Alzheimer and antioxidant activities of Coptidis Rhizoma
alkaloids,”Biol Pharm Bull, vol. 32 ,2009,pp. 1433-1438.
[43] Z. Slunska, E. Gelnarova, J. Hammerova, E. Tabrorska and I. Slainova,
“Effect of quaternary benzo(c)phenanthridine alkaloids sanguilutine and
chelilutine on normal and cancer cells,” Toxicol in vitro, vol. 24, 2010,
pp. 697-706. [44] R. Fayrer-Hosken, A. Stanley, N. Hill, G. Heusner, M. Christian, R. De
La Fuente, C. Baumann and L. Jones, “Effect of feeding fescue seed
containing ergot alkaloid toxins on stallion spermatogenesis and sperm
cells,” ReprodDomestAnim, vol. 47, 2012,pp. 1017-1026.
[45] R. Henkel, E. Kierspel, T. Stalf, C. Mehnert and R. Menkveld, “Effect of
reactive oxygen species produced by spermatozoa and leukocytes on
sperm functions in non-leukocytospermic patients,”FertilSteril, vol. 83,
2005, pp. 635-642.
[46] J. A. Martinez, A. Bello, L. L. Rubio, C. Rodriguez, L. Galan, E.
Caudales and J. L. Alvarez, “Calcium antagonist properties of the
bisbenzylisoquinoline alkaloid cycleanine,” FundamClinPharmacol, vol.
12, 1998, pp. 182-187.
[47] D. N. Guedes, J. M. Barbosa-Filho, V. S. Lemos and S. E. Cortes,
“Mechanism of the vasodilator effect of 12-O-methylcurine in rat aortic
rings,” J Pharm Pharmacol, vol. 54, 2002, pp. 853-858.
[48] B. T. Storey, C. L. Hourani and J. B. Kim, “A transient rise in
intracellular Ca2+ is a precursor reaction to the zonapellucida-induced
acrosome reaction in mouse sperm and is blocked by the induced
acrosome reaction inhibitor 3-quinuclidinyl benzilate,” MolReprodDev,
vol. 32, 1992, pp. 41-50.
[49] C. Gagnon and E. de Lamirande, “Controls of sperm motility. In: De
Jonge, C, Barratt C (Eds.), The Sperm Cell: Production, Maturation,
Fertilization and Regeneration,” Cambridge University Press, 2006,
Cambridge, UK, pp. 108–133.
[50] M. C. Chang, “Fertilizing capacity of spermatozoa deposited into the
fallopian tubes,” Nature J, vol. 168, 1951, pp. 697-698.
[51] C. R. Austin, “The capacitation of the mammalian sperm,” Nature J,
vol. 170, 1952, pp. 326.
[52] H. D. M. Moore and M. A. Akhondi,“Fertilizing capacity of rat
spermatozoa Is correlated With Decline in Straight-Line Velocity
Measured by Continuous computer-aided sperm analysis: epididymal rat
spermatozoa from the proximal cauda have a greater fertilizing capacity
in vitro than those from the distal cauda or vas deferens,” J Androl,
vol. 17, 1969, pp. 50-60.
[53] R. J. Aitken and K.M. West, “Analysis of the relationship between
reactive oxygen species production and leukocyte infiltration in fractions
of human semen separated on Percoll gradients,” Int J Androl, vol. 13,
1990, pp. 433-51.
[54] R. J. Aitken, E. Gordon and D. Harkiss, “Negative impact of oxidative
stress on the functional competence and genomic integrity of human
spermatozoa,” BiolReprod, vol. 59. 1998, pp. 1037-1046.
[55] R. J. Aitken and G. N. De Iuliis, “On the possible origins of DNA
damage in human spermatozoa,”Mol Hum Reprod, vol, 16, 2010,
pp. 3–13.
[1] N. E. Skakkebaek, N. Jorgensen, K. M. Main, E. Rajpert-De Meyts, H.
Leffers, A. Andersson, A. Juul, E. Carlsen, G. K. Mortensen, T. K.
Jensen, J. Toppari, “Human fecundity declining,”Int J Androl, vol. 29,
2006, pp.2-11.
[2] J. R. Sherins, “How is male fertility identified? In: Robaire B., Pryor J.
L., Trailer J.M. (eds.), Handbook of Andrology. American Society of
Andrology. Allen Press 1995, PP. 48-51.
[3] E. O. Laumann, A. Park and R. C. Rosen, “Sexual Dysfunction in
United States, Prevalence & Predictors,” JAMA, no. 6, 1999, pp. 281.
[4] H. G. Tempest, S. T. Homa, E. J. Routledge, A. Garner, X. P. and D. K.
Zhai, “Plants used in Chinese medicine for the treatment of male
infertility possess antioxidant and antioestrogenic activity,”
SystBiolReprod Med,VOL. 54, 2008, PP. 185-195.
[5] J. B. Calixto, “Efficacy, safety, quality control, marketing and regulatory
guidelines for herbal medicines (phytotherapeutic agents),”. Braz J Med
Biol Res, vol. 33, 2000, pp. 179-189.
[6] R. Yuan, and Y. Lin, “Traditional Chinese medicine: an approach to
scientific proof and clinical validation,” Pharmcol Therapeut, vol. 86,
2000, pp. 191-198.
[7] C. K. Christopher and M. T. Ho·Hui, “Rise of Herbal and Traditional
Medicine in Erectile Dysfunction management,” CurrUrol Rep, vol. 12,
2011, pp. 470-478.
[8] A. Adimoelja, “Phytochemicals and the breakthrough of traditional
herbs in the management of sexual dysfunctions,” Int J Androl, vol. 2,
2000, PP. 82-84.
[9] E. Nocerino, M. Amato and A. A. Izzo, “The aphrodisiac and
adaptogenic properties of ginseng,”Fitoterapia, vol. 71, pp. S1–S5.
[10] N. Erasmus, M. C. Solomom, K. A. Fortuin, and R. R. Henkel, “Effect
of Eurycomalongifolia Jack (Tongkatali) extract on human spermatozoa
in vitro,” Andrologia, vol. 44, 2012, pp. 308-314.
[11] C. Gonzales, J. Leiva-Revilla, J. Rubio, M. Gasco and G. F. Gonzales,
“Effect of red maca (Lepidium meyenii) on prostate zinc levels in rats
with testosterone-induced prostatic hyperplasia,” Andrologia, vol. 44,
2012, pp. 362-369.
[12] P. F. Moundipa, S. Ngouela, P. Kamtchouing, E. Tsamo, F. M.
Tchouanguep and S. Carreau,“Effects of extracts from Hibiscus
macranthus and Basella alba mixture on testosterone production in vitro
in adult rat testes slices,” Asian J Androl, vol. 8, 2006, pp. 111-114.
[13] F. Lampiao, D. Krom and S. S. du Plessis, “The in vitro effects of
Mondia whitei on human sperm motility parameters,” Phytother Res vol.
22, 2008, pp.1272-1273.
[14] J. J. Meyer Marian, C. N. Rakuambo and A. A. Hussen, “Novel
xanthones from Securidacalongependunculata with activity against
erectile dysfunction,” JEthnopharmacol, vol. 119, 2008, pp. 559-603.
[15] E. E. van Wyk, B. van Oudtshoorn and N. Gericke,“Medicinal Plants of
South Africa,” 2nd edition; Briza Publications 2009, Pretoria, South
Africa.
[16] B. E. van Wyk and N. Gericke, “People’s Plants. A Guide to the Useful
Plants of Southern Africa,” Briza 2000, Pretoria, South Africa.
[17] B. E. Van Wyk, F. R. van Heerden and B. van Oudtshoorn, “Poisonous
Plants of South Africa,” Briza Publications 2002, Pretoria, South Africa.
[18] H. De Wet and B. E. van Wyk, “An ethno-botanical survey of southern
African Menispermaceae,”S Afr J Bot, vol. 74, 2008, pp. 2-9.
[19] M. J. Watt and M. G. Breyer-Brandwijk, “The medicinal and poisonous
plants of southern and eastern Africa,” 2nd edition. Livingston, London.
[20] C. A. Smith “Common names of South African plants,” Memoirs of the
botanical survey of South Africa, no. 35, 1966, department of agriculture
and technical services.
[21] E. Von Koenen, “Medicinal, Poisonous, and Edible Plants of Namibia,”
Klaus Hess Publishers/Verlag, Windhoek 2001, Namibia.
[22] S. M. Shalaweh, N. Erasmus, F. Weitz and R. R. Henkel, “Effect of
Cissampelos capensis rhizome extract on human spermatozoa in vitro,”
Andrologia, doi: 10.1111/and.12264,2014, pp. 1-10.
[23] R. Henkel, E. Kierspel, M. Hajimohammad, T. Stalf, C. Hoogendijk, C.
Mehnert, R. Menkveld, W. Schill and T. F. Kruger, “DNA
fragmentation of spermatozoa and assisted reproduction technology,”
Reprod Biomed Online, vol. 7,2013, pp. 477-484.
[24] R. Henkel, M. Hajimohammad, T. Stalf, C. Hoogendijk, C. Mehnert, R.
Menkveld, H. Gips, W. B. Schill and T. F. Kruger, “Influence of
deoxyribonucleic acid damage on fertilization and pregnancy,”
FertilSteril, vol. 81, 2004,pp. 965-672.
[25] W. Gorczyca, F. Traganos, H. Jesionowska and Z. Daryznkiewcz,
“Presence of strand breaks and increased sensitivity of DNA in situ to
denaturation in abnormal human sperm cells: analogy to apoptosis in
somatic cells,”Exp Cell Res, vol. 207, 2003, pp. 202-205.
[26] C. M. Green, S. M. Cockle, P. F. Watson and L. R. Fraser, “Fertilization
promoting peptide, a tripeptide similar to thyrotrophin-releasing
hormone, stimulates the capacitation and fertilizing ability of human
spermatozoa in vitro,” Hum Reprod, vol. 11, 1996, pp. 830-836.
[27] WHO, “Global Atlas of Traditional, Complementary and Alternative
Medicine,” vol. 1 and 2, World Health Organization 2005, Geneva.
[28] D. S. Fabricant and N. R. Farnsworth, “The value of plants used in
traditional medicine for drug discovery,” Environmental health
perspectives, vol. 109, 2001, pp. 69.
[29] H. Azaizeh, S. Fulder, K. Khalil and O. Said, “Ethnobotanical
knowledge of local Arab practitioners in the Middle Eastern region,”
Fitoterapia,vol. 74, 2003, pp. 98-108.
[30] R. A. Halberstein, “Medicinal plants: historical and cross-cultural usage
patterns,” Annals of epidemiology, vol. 15, 2005, pp. 686-699.
[31] P. J. Houghton, “The role of plants in traditional medicine and current
therapy,” J & C M, vol. 1, 1995, pp. 131-143.
[32] R. Yuan and Y. Lin, “Traditional Chinese medicine: an approach to
scientific proof and clinical validation,” Pharmacology & therapeutics,
vol. 86, 2000,pp. 191-198.
[33] M. Solomon and R. R. Henkel, “In vivo effects of Eurycomalongifolia
Jack (Tongkat Ali) extract on reproductive functions in the
rat,”Andrologia, vol. 46 ,2014, pp. 339-348.
[34] H. De Wet, F. R. van Heerden and B. E. van Wyk, “Alkaloidal variation
in Cissampeloscapensis (Menispermaceae),” Molecules, vol. 16, 2011,
pp. 3001-3009.
[35] E. DeLamirande, P. Leclerc and C. Gagnon, “Capacitation as a
regulatory event that primes spermatozoa for the acrosome reaction and
fertilization,” Mol. Hum. Reprod, vol. 3, 1997,pp. 175–194.
[36] P. Olds-Clarke, “Unresolved issues in mammalian fertilization,”Int. Rev.
Cyt, vol, 232, 2003, pp. 129–184.
[37] C. De Jonge, “Biological basis for human capacitation,” Hum. Reprod,
vol. 11, 2005, pp. 205–214.
[38] C. O’Flaherty, E. de Lamirande and C. Gagnon, “Positive role of
reactive oxygen species in mammalian sperm capacitation triggering and
modulation of phosphorylation events, Free Radic,”Biol. Med, vol. 41,
2006, pp. 528–540.
[39] E. DeLamirande and C. O'Flaherty, “Sperm activation: role of reactive
oxygen species and kinases,” Biochim Biophys Acta, vol. 1, 2008, pp.
106-115.
[40] B. Zhao, Y. Chen, X. Sun, M. Zhou, J. Ding, J. J. Zhan andL. J. Guo,
“Phenolic alkaloids from Menispermum dauricum rhizome protect
against brain ischemia injury via regulation of GLT-1, EAAC1 and ROS
generation,” Molecules, vol. 17, 2012, pp. 2725-2737.
[41] W. H. Chiu, S. J. Luo, C.L Chen, J. H. Cheng, C. Y. Hsieh, C. Y. Wang,
Huang, W. C. Su and C. F. Lin, “Vinca alkaloids cause aberrant ROSmediated
JNK activation, Mcl-1 downregulation, DNA damage,
mitochondrial dysfunction, and apoptosis in lung adenocarcinoma cells,”
Biochem Pharmacol, vol. 83, 2012,pp. 1159-1171.
[42] H. A. Jung, B. S. Min, T. Yokozawa, J. H. Lee, Y. S. Kim and J. S.
Choi, “Anti-Alzheimer and antioxidant activities of Coptidis Rhizoma
alkaloids,”Biol Pharm Bull, vol. 32 ,2009,pp. 1433-1438.
[43] Z. Slunska, E. Gelnarova, J. Hammerova, E. Tabrorska and I. Slainova,
“Effect of quaternary benzo(c)phenanthridine alkaloids sanguilutine and
chelilutine on normal and cancer cells,” Toxicol in vitro, vol. 24, 2010,
pp. 697-706. [44] R. Fayrer-Hosken, A. Stanley, N. Hill, G. Heusner, M. Christian, R. De
La Fuente, C. Baumann and L. Jones, “Effect of feeding fescue seed
containing ergot alkaloid toxins on stallion spermatogenesis and sperm
cells,” ReprodDomestAnim, vol. 47, 2012,pp. 1017-1026.
[45] R. Henkel, E. Kierspel, T. Stalf, C. Mehnert and R. Menkveld, “Effect of
reactive oxygen species produced by spermatozoa and leukocytes on
sperm functions in non-leukocytospermic patients,”FertilSteril, vol. 83,
2005, pp. 635-642.
[46] J. A. Martinez, A. Bello, L. L. Rubio, C. Rodriguez, L. Galan, E.
Caudales and J. L. Alvarez, “Calcium antagonist properties of the
bisbenzylisoquinoline alkaloid cycleanine,” FundamClinPharmacol, vol.
12, 1998, pp. 182-187.
[47] D. N. Guedes, J. M. Barbosa-Filho, V. S. Lemos and S. E. Cortes,
“Mechanism of the vasodilator effect of 12-O-methylcurine in rat aortic
rings,” J Pharm Pharmacol, vol. 54, 2002, pp. 853-858.
[48] B. T. Storey, C. L. Hourani and J. B. Kim, “A transient rise in
intracellular Ca2+ is a precursor reaction to the zonapellucida-induced
acrosome reaction in mouse sperm and is blocked by the induced
acrosome reaction inhibitor 3-quinuclidinyl benzilate,” MolReprodDev,
vol. 32, 1992, pp. 41-50.
[49] C. Gagnon and E. de Lamirande, “Controls of sperm motility. In: De
Jonge, C, Barratt C (Eds.), The Sperm Cell: Production, Maturation,
Fertilization and Regeneration,” Cambridge University Press, 2006,
Cambridge, UK, pp. 108–133.
[50] M. C. Chang, “Fertilizing capacity of spermatozoa deposited into the
fallopian tubes,” Nature J, vol. 168, 1951, pp. 697-698.
[51] C. R. Austin, “The capacitation of the mammalian sperm,” Nature J,
vol. 170, 1952, pp. 326.
[52] H. D. M. Moore and M. A. Akhondi,“Fertilizing capacity of rat
spermatozoa Is correlated With Decline in Straight-Line Velocity
Measured by Continuous computer-aided sperm analysis: epididymal rat
spermatozoa from the proximal cauda have a greater fertilizing capacity
in vitro than those from the distal cauda or vas deferens,” J Androl,
vol. 17, 1969, pp. 50-60.
[53] R. J. Aitken and K.M. West, “Analysis of the relationship between
reactive oxygen species production and leukocyte infiltration in fractions
of human semen separated on Percoll gradients,” Int J Androl, vol. 13,
1990, pp. 433-51.
[54] R. J. Aitken, E. Gordon and D. Harkiss, “Negative impact of oxidative
stress on the functional competence and genomic integrity of human
spermatozoa,” BiolReprod, vol. 59. 1998, pp. 1037-1046.
[55] R. J. Aitken and G. N. De Iuliis, “On the possible origins of DNA
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@article{"International Journal of Biological, Life and Agricultural Sciences:70225", author = "S. Shalaweh and P. Bouic and F. Weitz and R. Henkel", title = "Cissampelos capensis Rhizome Extract Induces Intracellular ROS Production, Capacitation and DNA Fragmentation in Human Spermatozoa", abstract = "More than 3000 plants of notable phyto-therapeutic
value grow in South Africa; these include Cissampelos capensis,
commonly known in Afrikaans as dawidjie or dawidjiewortel. C.
capensis is the most significant and popular medicinal plant used by
the Khoisan as well as other rural groups in the Western region of
South Africa. Its rhizomes are traditionally used to treat male fertility
problems. Yet, no studies have investigated the effects of this plant or
its extracts on human spermatozoa. Therefore, this study aimed at
investigating the effects of C. capensis rhizome extract (CRE)
fractions on ejaculated human spermatozoa in vitro. Spermatozoa
from a total of 77 semen samples were washed with human tubular
fluid medium supplemented with bovine serum albumin (HTF-BSA)
and incubated for 2 hours with 20 μg/ml progesterone (P4) followed
by incubation with different concentrations (0, 0.05, 0.5, 5, 50, 200
μg/ml) of fractionated CRE (F1=0% MeOH, F2=30% MeOH,
F3=60% MeOH and F4=100% MeOH) for 1.5 hours at 37°C. A
sample without addition of CRE fractions served as control. Samples
were analyzed for sperm motility, reactive oxygen species (ROS),
DNA-fragmentation, acrosome reaction and capacitation. Results
showed that F1 resulted in significantly higher values for ROS,
capacitation and hyper-activation compared to F2, F3, and F4 with
P4-stimulated samples generally having higher values. No significant
effect was found for the other parameters. In conclusion, alkaloids
present in F1 of CRE appear to have triggered sperm intrinsic ROS
production leading to sperm capacitation and acrosome reaction
induced by P4.", keywords = "Capacitation, acrosome reaction, Cissampelos
capensis, DNA fragmentation, ROS.", volume = "9", number = "5", pages = "576-8", }
