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The effects of Cecropia obtusifolia and Cecropia polyphlebia on lung capacity in relation to alkaloid concentration Mai Lee Xiong Department of Microbiology, University of Wisconsin Madison ABSTRACT For centuries, people have depended on nature t o alleviate their ailments. A popular natural remedy for asthma is the use of Cecropia adenopus (Rivero 2008). In this study, I looked at two Cecropia spp., C. obtusifolia and C. polyphlebia found in Monteverde, Costa Rica. Cecropia obtusifolia is foun d at lower elevation and has a mutualism with Azteca ants while C. polyphlebia is found at higher elevation in the Cloud Forest with no ant mutualism. A previous study by Ross (2003) found that ingestion of C. obtusifolia leaf decoction increased lung cap acity by three to four percent compared to a placebo group. A possible reason is the presence of alkaloids (Schmeller & Wink 1998) in leaves of Cecropiaceae (Morton 1981). I predicted that since C. obtusifolia has a mutualism with ants, it would have les s alkaloids compared to C. polyphlebia thus rendering it less effective as an anti asthmatic agent. I followed the protocols from Ross (2003) for tea preparation and lung capacity measurement and Gottheiner (1998) for alkaloid extraction. I found no inc rease in lung capacity from drinking Cecropia decoctions. The mean rate of change, which is the final lung capacity measurement minus the initial divided by the number of samples in a group, for the control is 0.136 L with a standard error of 0.066; C. ob tusifolia 0.180 L and 0.090; and C. polyphlebia 0.172 L and 0.100 (one way ANOVA; F 2,52 = 0.056, p = 0.946). Further, while C. polyphlebia may have more alkaloids, this was not significant enough to make a difference in lung capacity. RESUMEN Por cen turias los seres humanos han dependido de la naturaleza para aliviar sus males. Un remedio popular para el asma es el uso de Cecropia adenopus. Estudie Cecropia obtusifolia y Cecropia polyphlebia en Monteverde, Costa Rica. Cecropia obtusifolia se encuent ra a elevaciones mas bajas y tiene un mutualismo con hormigas Azteca mientras que C. polyphlebia se encuentra a mayores alturas sin mutualismo con las hormigas. Un estudio previo encontr que las infusiones de C. obtusifolia incrementan la capacidad pulm onar entre un tres y cuatro por ciento comparado con un placebo, probablemente por la precencia de alcaloides. Debido a que C. polyphebia no tiene un mutualismo con hormigas deberia tener una mayor concentracin de alcaloides, por lo tanto un mayor efecto antiasmatico. A pesar de que se encontr una mayor concentracin de alcaloides en C. polyphebia no se encontraron diferencias en el efecto de los tratamientos en la capacidad pulmonar. INTRODUCTION The World Health Organization (WHO 2008) estimated t hat 80% of the population in some Asian and African countries depends on traditional medicine, probably due to the fact that more than one billion people do not have access to health care services and modern medicine (Carr 2004). In Belize, at least 75% o f its population receives primary health care from traditional practitioners (Balick & Mendelsohn 1992), and this may well be true for most developing countries. For ailments, like asthma, where there are an estimated 300 million people suffering, and is the most common chronic disease for children (WHO 2008) a cheap, readily available traditional cure is important for people in the developing world.
For centuries, humans have used the Cecropia trees as herbal remedies to alleviate asthma (Raintree Nutri tion 2007). Cecropia trees are prevalent in the Neotropics, growing in disturbed areas such as roadside, forest edge, and landslide easily accessible for humans to use. Of particular interest are the two Cecropia species in the Monteverde area, C. obtusi folia and C. polyphlebia Cecropia obtusifolia has been shown to control hypertension (Salas et al. 1987; Lima Landman et al. 2007), hypoglycemia (Roman Ramos et al. 1991; Andrade Cetto & Wiedenfeld 2001), and act as a diuretic (Vargas and Montero 1996), as well as an analgesic depressor (Perez Guerrero et al. 2001). Ross (2003) found that consuming C. obtusifolia leaf decoctions increased lung capacity by three to four percent compared to a placebo group. This is the first clinical demonstration of the pharmacological potential of Cecropia spp. for asthma, bronchitis, and other respiratory diseases. A set of important compounds shown to have medicinal value are alkaloids. Schmeller and Wink (1998) reported on the different uses of alkaloid compounds as painkillers, stimulants and hallucinogens by humans. In nature, alkaloids are secondary compounds, known to be produced by plants as a chemical defense against herbivores, microorganism, viruses, and other plants (Schmeller & Wink, 1998). Although C. o btusifolia contains alkaloid, flavonoids, tannins (Morton 1981) and other compounds, its main defense against herbivores is a mutualistic relationship with Azteca ants. C. polyphlebia growing at higher altitudes, has lost its ant partner and should have a higher concentration of alkaloids and other secondary compounds in its leaves to compensate (Scalley 1993). Because Cecropia spp have been used traditionally to control asthma and Ross (2003) found C. obtusifolia to increase lung capacity, it is likely that the more chemically protected C. polyphlebia would do an even better job. Therefore, this study examines if ingestion of leaf decoction of Cecropia polyphlebia will have a higher increase in lung capacity compare to C. obtusifolia and determines if C. polyphlebia contains higher level of alkaloids. METHODS Cecropia obtusifolia and C. polyphlebia were identified by location (altitude), number of leaflets, length of inflorescences, and the presence (or absence) of ant mutualism, following guidelin es by Zuchowski (2007) and Longino (2005). Cecropia obtusifolia an ant mutualistic plant, occurs in disturbed areas of moist to wet forest from 1000 to 1500 m elevation, and has 10 13 long leaflets with inflorescence spikes to more than 50 cm long. In contrast, C. polyphlebia has no mutualism with ants, short inflorescence spikes less than 6 cm long, 10 11 leaflets, and occurs above 1500 m in the Cloud Forest. Study sites The leaves of C. obtusifolia were gathered from four individual plants on r oadsides near Centro Panamericano de Idiomas (CPI) Monteverde and El Hotel de Montana (1350 m). Cecropia polyphlebia leaves were collected in the Monteverde Cloud Forest near the Biological Station of Monteverde (1550 m). Cecropia Tea and Lung Capacity Experiment
I collected one young leaf per individual plant for a total of four individuals for each species. The leaves were boiled and made into tea according to Ross (2003) with the exception that I increased the amount of leave tissue per one liter of water from two grams to four grams. The placebo tea was made from diluted chamomile ( Matricaria recutita : Asteraceae) tea. I made the teas in bulk and assigned treatments randomly to participants on the same day each participant to one treatment. A t otal of 55 volunteers participated, 18 for each of the treatment groups and 19 for the control. The average participant was a 21 year old college female who exercised at least twice a week and had no pre existing respiratory diseases. The participants we re instructed to refrigerate the tea and drink 100 mL of tea after breakfast for five days. I measured the lung capacity of an individual by having the person blow a balloon in one breath, and this was completed before the individual had consumed the tea as the initial lung capacity measurement. Another lung capacity measurement was taken after the fifth day as the final lung capacity. The volume of the balloon, and thus the lung capacity of the participant, was determined through water displacement. Alkaloid Extraction from C. obtusifolia and C. polyphlebia I completed the same leaf collection for my alkaloid extraction. The species were collected and dried in a drying box for a day. Then, the dried leaves were ground and soaked in methanol overnig ht. The rest of the alkaloid extraction procedures were from Gottheiner (1998). All the samples were analyzed with a MRC Spectrophotometer, Model UV 200 RS at 540 nm, and measured in percent transmittance (% T). RESULTS There was no statistical signifi cant between the treatment groups and placebo group even though the alkaloid concentration of C. polyphlebia was slightly higher than C. obtusifolia Cecropia Tea Lung Capacity Experiment Comparing measurements of the initial and final lung capacity, I found no difference between Cecropia teas and the placebo chamomile tea (one way ANOVA; F 2,52 = 0.056, p = 0.946). Figure one shows the mean rate of change (L) which is the final lung capacity measurement minus the initial divided by the number of samples in each group for each treatment with the standard errors. The rate of change for the control is 0.136 L with a standard error of 0.066; C. obtusifolia 0.180 L and 0.090; and C. polyphlebia 0.172 L and 0.100. Although Cecropia obtusifolia and C. polyph lebia have a slightly higher rate of change compared to the control, the overlap of the standard errors indicates that there is no statistical difference between the two treatment groups and the placebo group.
Alkaloid Extraction of C. obtusifol ia and C. polyphlebia The percent transmittance (% T) for Cecropia obtusifolia was 97 and for C polyphlebia 95. This shows that C. polyphlebia have a slightly higher concentration of alkaloid because the C. polyphlebia sample absorbed more light. How ever, the exact alkaloid concentration is not known because a standard curve was not generated. DISCUSSION My experiment examined the effects of ingestion of Cecropia obtusifolia and C. polyphlebia leaf decoction on lung capacity. I expected to find th at C. polyphlebia would have a higher increase in lung capacity due to the fact that it lacks ants, and thus, it likely increases its synthesis of alkaloids and other secondary compounds for protection against herbivores and pathogens. My results indicate d that while C. polyphlebia contained a slightly higher concentration of alkaloid than C. obtusifolia there was no difference in its impact on lung capacity. In fact, neither Cecropia species increased lung capacity compared to the control. This was a s urprising result since a previous student investigator found a three to four percent lung capacity increase using C. obtusifolia both in short time effectiveness (measured one minute after leaf ingestion) and long term effectiveness (measured after five da ys), respectively (Ross Figure 1. The average rate of change for each treatment group Cecropia obtusifolia and C. polyphlebia have a slightly higher rate of change compared to the control group but it is not statistically significant.
2003). I even doubled the strength of the decoction by doubling the grams of leaves per liter of water from two grams (Ross 2003) to four grams, expecting to find a subsequent increase in lung capacity. While I cannot explain why Ross (2003) found different result, an herbal medicine website (Rivero 2008) and a scientific paper (Alonso 1998) recommend Cecropia leaves decoction to be 20 40 grams per one liter of water for the treatment of asthma. The expected effect is one hour a fter tea consumption and should be consumed two to four times daily. Clearly, this is an area that needs to be explored in more depth. For future studies, it is pertinent to have better lung capacity measuring equipment as this will insure increased a ccuracy. Any decoction of Cecropia leaves should follow the recommended amount of 20 40 grams per one liter of water and should be taken at least twice a day. In addition, a standard curve should be generated to calculate the alkaloid concentration of Cecropia obtusifolia and C. polyphlebia This may validate the need to decoct more leaves per one liter of water. Lastly, the importance of Cecropia trees and its effect on asthma cannot be ignored. The discovery of such treatment would mean that million s of sufferers of asthma or other respiratory diseases would benefit tremendously, especially when access to health care services is limited. ACKNOWLEDGEMENTS I want to thank Alan and Karen Masters for their guidance and support you guys rock my world! A thank goes to Jose (Moncho) Calderon for his help in getting the Cecropia leaves where my short stature cannot reach and my materials and equipments; and also, to Yi men Araya for always being a friend and helping me any way possible that he can. I want to acknowledge the CIEE, EAP and SE program, professors, TAs and the cooks (also, for the use of the refrigerator) for their participation as tea takers for my experiment. An enormous thanks to Virginia, my homestay grandma, for the use of her kitchen an d kitchen utensils and for putting up with me when I was up at two o'clock in the morning making my tea. And lastly, thanks to anyone that I might have missed, I just want to say that even if the help was small it helped in aiding the progression of my pr oject. LITERATURE CITED Alonso, J. 1998. Tratado de Fitomedicina. Bases clinicas y farmacologicas. Isis Ediciones, Buenos Aires. 258 as found in Consolini, A.E. & Migliori, G.N. 2005. Cardiovascular effects of the South American medicinal plant Cecro pia pachystachya (ambay) on rats. J of Ethnopharm 96: 417 422 Andrade Cetto, A. & Wiedenfeld, H. 2001. Hypoglycemic effect of Cecropia obtusifolia on streptozotocin diabetic rats. J of Ethnopharm 78(2 3): 145 49 Balick, M.J. & Mendelsohn, R. 1992. Assess ing the economic value of traditional medicines from tropical rain forests. Conservation Biology 6(1): 128 30 Carr, D. 2004. Improving the Health of the World's Poorest People in Health Bulletin. Population Reference Bureau. 12 May 2009. http://www.prb.org/pdf/ImprovingtheHealthWorld_Eng.pdf Consolini, A.E. & Migliori, G.N. 2005. Cardiovascular effects of the South American medicinal plant Cecropia pachystachya (ambay) on rats. J of Ethn opharm 96: 417 422 Consolini, A.E., Ragone, M.I., Migliori, G.N., Conforti, P., Volonte, M.G. 2006. Cardiotonic and sedative effects on Cecropia pachystachya Mart. (ambay) on isolated rat hearts and conscious mice. J of Ethnoparmacology 106:90 96 Gotthein er, D.M. Fall 1998. An analysis of the relationship between alkaloids and herbivory in Cecropia obtusifolia and Cecropia polyphlebia EAP Fall 1998 Lima Landman, M.T.R., Borges, A.C.R., Cysneiros, R.M., De Lima, T.C.M., Souccar, C., Lapa, A.J. 2007. Antihy pertensive effect of a standardized aqueous extract of Cecropia glaziovii Sneth in rats: An in vivo approach to the hypotensive mechanism. Phytomedicine 14:314 320.
Longino, J.T. 2005. The Cecropia Azteca association in Costa Rica. 12 May 2009. http://academic.evergreen.edu/projects/ants/ANTPLANTS/CECROPIA/Cecropia.html Rivero, R. 2008. Treatment of the asthma with Cecropia adenopus. Mama Herb. 15 May 2009. http://www.mamaherb.com/Treatments/View.aspx?id=5878&conditionId=0&specificConditionId=62 Morton, J. 1987. Atlas of Medicinal Plants of Middle America Charles C. Thomas Publishers, USA (1981) as found in Perez Guerrero, C., Herrera, M.D., Ortiz, R., Alvarez de Sotomayor, M., Fernandez, M.A., 2001. A pharmacological study of Cecropia obtusifolia Bertol aqueous extract. J of Ethnopharm 76:279 284. Perez Guerrer o, C., Herrera, M.D., Ortiz, R., Alvarez de Sotomayor, M., Fernandez, M.A., 2001. A pharmacological study of Cecropia obtusifolia Bertol aqueous extract. J of Ethnopharm 76:279 284. Raintree Nutrition, Inc. 2007. Tropical Plant Database: embauba. 15 May 2009. http://www.rain tree.com/cecropia.htm Roman Ramos, R., Flores Saenz, J.L., Partida Hernandez, G., Lara Lemus,A., Alarcon Aguilar, F., 1991. Experimental study of the hypoglycemic effect of some anti diabetic plants. Archivos de Investigacion Medica (Mexico) 22:87 93. Ross, C. Fall 2003. Effects of Cecropia obtusifolia leaf ingestion on lung capacity. Tropical Ecology and Conservation p. 207 Schmeller, T. & Wink, M. 1998. Utilization of alkaloids in m odern medicine. Alkaloids: Biochemistry, Ecology, and Medicinal Applications Roberts & Wink (eds). Institute for Pharmaceutical Biology, University of Heidelberg, Germany. Plenum Press, New York. p. 435 (book from web) Salas, Y., Brenes, J.R., Morales, O. M., 1987. Antihypertensive effect of Cecropia obtusifolia (Moraceae) leaf extract on rats Revista de Biologa del Tropico 35:127 130 Vargas Howell, R., Ulate Montero, G., 1996. Diuretic effect of Cecropia obtusifolia (Moraceae) on albino rats. Revista de Biologa del Tropico 44:93 96. World Health Organization (WHO). 2008. Traditional Medi cine 12 May 2009. http://www.who.int/mediacentre/factsheets/fs/34/en/index.html Zuchowski, W. 2005. A Guide to Tropical Plants of Costa Rica A Zona Tropical Publication.
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Xiong, Mai Lee
Los efectos de Cecropia obtusifolia y Cecropia polyphlebia en la capacidad pulmonar en relacin con la concentracin de alcaloides
The effects of Cecropia obtusifolia and Cecropia polyphlebia on lung capacity in relation to alkaloid concentration
For centuries, people have depended on nature to alleviate their ailments. A popular natural remedy for asthma is the use of Cecropia adenopus (Rivero 2008). In this study, I looked at two Cecropia spp., C. obtusifolia and C. polyphlebia, found in Monteverde, Costa Rica. Cecropia obtusifolia is found at lower elevation and has a mutualism with Azteca ants while C. polyphlebia is found at higher elevation in the Cloud Forest with no ant mutualism. A previous study by Ross (2003) found that ingestion of C. obtusifolia leaf decoction increased lung capacity by three to four percent compared to a placebo group. A possible reason is the presence of alkaloids
(Schmeller & Wink 1998) in leaves of Cecropiaceae (Morton 1981). I predicted that since C. obtusifolia has a mutualism with ants, it would have less alkaloids compared to C. polyphlebia, thus rendering it less effective as an anti-asthmatic agent. I followed the protocols from Ross (2003) for tea preparation and lung capacity measurement and Gottheiner (1998) for alkaloid extraction. I found no increase in lung capacity from drinking Cecropia
decoctions. The mean rate of change, which is the final lung capacity measurement minus the initial divided by the number of samples in a group, for the control is 0.136 L with a standard error of 0.066; C. obtusifolia, 0.180 L and 0.090; and C. polyphlebia, 0.172 L and 0.100 (one-way ANOVA; F2,52 = 0.056, p = 0.946). Further, while C. polyphlebia may have more alkaloids, this was not significant enough to make a difference in lung capacity.
Durante siglos los seres humanos han dependido de la naturaleza para aliviar sus dolencias. Un remedio natural popular para el asma es el uso de Cecropia adenopus. Estudie Cecropia obtusifolia y Cecropia polyphlebia en Monteverde, Costa Rica.
Text in English.
Tropical Ecology 2009
Ecologa Tropical 2009
t Monteverde Institute : Tropical Ecology