EVALUATION OF PHYTOCHEMICAL COMPOSITION OF PLANTS OF GENUS HAWTHORN (CRATAEGUS L.) GROWING IN LITHUANIA INTRODUCTION For the last fifteen years, the attention of doctors and their patients on drugs and food supplements of herbal origin has been continuously increasing. The growing interest in raw plant materials is expanding the possibilities of use of these materials in the industries of medicine, cosmetics and food as well as encouraging assessment and description of natural resources using new modern methods. Scientific research on natural herbal resources has bee receiving special attention in the European Union. In recent years, a number of important documents have been published in the European Union, which declare the aims of ensuring the use and quality of natural herbal resources, such as Good Agricultural Practice document, Points to consider on Good agricultural and collection practice for starting materials of herbal origin, a long term vision of the future of herbal biotechnology in Europe Plants for the Future. Cardiovascular system diseases constitute one of the most urgent health problems. The range of effective herbal preparations for the target phytotherapy of cardiovascular system diseases is not wide. Hawthorn preparations are a promising drug for the treatment of cardiovascular system diseases, whose pharmacological effects have been substantiated by scientific studies. Throughout the territory of Lithuania, three kinds of hawthorn can be found growing... [to full text]

Tyrimo tikslas: įvertinti bioaktyvių junginių kiekybinės sudėties bei antioksidantinio aktyvumo įvairavimą vienapiestės gudobelės žaliavų mėginiuose, rinktuose skirtinguose Lietuvos vietovėse. Tyrimo uždaviniai: Įvertinti suminio flavonoidų kiekio ir suminio fenolinių junginių kiekio įvairavimą vienapiestės gudobelės vaisių ir lapų mėginiuose, rinktuose skirtingose Lietuvos vietovėse; įvertinti chlorogeno rūgšties ir flavonoidų (epikatechino, hiperozido, izokvercitrino, viteksino-2“-O-ramnozido, rutino) kiekinį pasiskirstymą C.monogyna žaliavose efektyviosios skysčių chromatografijos metodu; ištirti gudobelių vaisių ir lapų žaliavų antioksidantinį aktyvumą ir įvertinti jo įvairavimą naudojant DPPH ir ABTS metodus; pritaikyti klasterinę analizę vienapiesčių gudobelių vaisių ir lapų mėginių grupavimui, iškiriant reikšmingai besiskiriančias antioksidantiniu aktyvumu grupes. Tyrimo metodai: Flavonoidų kiekiui įvertinti naudoti spektrofotometriniai ir efektyviosios skysčių chromatografijos metodai, suminis fenolinių junginių kiekis įvertintas spekrofotometriškai. Antioksidantinis aktyvumas nustatytas spektrofotometriniu metodu, naudojant DPPH ir ABTS testus. Tyrimo objektas: Lietuvos skirtinguose regionuose rinktos Crataegus monogyna Jacq. vaisių ir lapų žaliavos. Tyrimo rezultatai: Flavonoidų kiekis lapų žaliavose įvairavo nuo 0,47 iki 2,34 proc., vaisių žaliavoje nuo 0,12 iki 0,91proc.. Suminis fenolinių junginių kiekis lapų mėginiuose nuo 5,68 iki 11,80 proc., vaisių... [toliau žr. visą tekstą] / The aim of the research: to evaluate the variety of quantitative composition of bioactive combination and antioxidant activity in the samples of C. monogyna selected in different parts of Lithuania. The objectives of the research: to evaluate the variety of total amount of flavonoids and total amount of phenolic combinations in the samples of C. monogyna leaves and fruit; to evaluate the quantitative distribution of of chlorogenic acid and flavonoids in C. monogyna raw materials by the method of effective chromatography of liquid; to analyse the antioxidant activity of hawthorn leaves and fruit and evaluate the variety of it using DPPH and ABTS methods; to apply cluster analysis of sample grouping of monopistil howthorn leaves and fruit marking out groups having definetely different level of antioxidant activity. The methods of the research: the methods of spectrophotometry and efective liquid chromatography were used to evaluate the amount of flavonoids and the total amount of phenolic combinations. The antioxidant activity was established using DPPH and ABTS tests. The object the research: Crataegus monogyna Jacq. raw material of leaves and fruit selected in different parts of Lithuania. The results of the research: the amount of flavonoids in leaves differed from 0,47 to 2,34%; in fruit it difffered from 0,12 to 0,91 %. Total amount of phenolic combinations in the samples of leaves differed from 5,68 to 11,80 percent; in raw material of fruit differed from 1,03 to 2,78%... [to full text]

Polyploidization is exceptionally cosmopolitan in plants. One common observation is that polyploids inhabit larger geographic distributions than their diploid progenitors. Differences in distribution between cytotypes are largely attributed to differences in mating system and ecological breadth among cytotypes. In Crataegus series Douglasianae, allopolyploids have larger ranges than their diploid progenitors. Range size increase is coupled with a shift to predominant asexuality in polyploids. This thesis explores 2 additional hypotheses that may contribute to differences in distribution among cytotypes: ecological breadth and dispersal ability. We find evidence that tetraploid C. douglasii occurs in a wider range of habits and has a greater dispersal ability than diploid C. suksdorfii. Overall, we suggest that differences in mating system, ecological breadth, and dispersal ability have contributed individually and collaboratively to differences in distribution among cytotypes of Crataegus series Douglasianae. Large ranges in polyploids may help maintain cytotype diversity by providing buffering capacity against demographic stochasticity.

Polyploidization is exceptionally cosmopolitan in plants. One common observation is that polyploids inhabit larger geographic distributions than their diploid progenitors. Differences in distribution between cytotypes are largely attributed to differences in mating system and ecological breadth among cytotypes. In Crataegus series Douglasianae, allopolyploids have larger ranges than their diploid progenitors. Range size increase is coupled with a shift to predominant asexuality in polyploids. This thesis explores 2 additional hypotheses that may contribute to differences in distribution among cytotypes: ecological breadth and dispersal ability. We find evidence that tetraploid C. douglasii occurs in a wider range of habits and has a greater dispersal ability than diploid C. suksdorfii. Overall, we suggest that differences in mating system, ecological breadth, and dispersal ability have contributed individually and collaboratively to differences in distribution among cytotypes of Crataegus series Douglasianae. Large ranges in polyploids may help maintain cytotype diversity by providing buffering capacity against demographic stochasticity.

Herbs have been used for many centuries in diverse civilizations for the treatment of heart disease. Only a few natural supplements claim to have direct cardiovascular actions including hawthorn (Crataegus spp.) and berberine derived from the Berberidaceae family. Several different studies indicate important cardiovascular effects of hawthorn and berberine. For example, both exert positive inotropic effects and have been used in the treatment of congestive heart failure. Recently, it was shown that hawthorn extract preparations cause negative chronotropic effects in a cultured neonatal murine cardiomyocyte assay independent of beta-adrenergic receptor blockade. The aim of this study was to further characterize the effect of hawthorn extract to decrease the contraction rate of cultured cardiomyocytes. We hypothesized that hawthorn extract may be acting through muscarinic receptors to decrease contraction rate of cardiomyocytes. Atrial and ventricular cardiomyocytes were treated with hawthorn extract in the presence of atropine or himbacine. Changes in the contraction rate of cultured cardiomyocytes revealed that both muscarinic antagonists significantly attenuated the negative chronotropic activity of hawthorn extract. Using quinuclidinyl benzilate, L-[benzylic-4,4'-3H] ([³H]-QNB) as a radioligand antagonist, the effect of a partially purified hawthorn extract fraction to inhibit muscarinic receptor binding was quantified. Hawthorn extract fraction 3 dose-dependently inhibited [³H]-QNB binding to mouse heart membranes. These findings suggest that muscarinic receptors may be involved in the negative chronotropic effect of hawthorn extracts in neonatal murine cardiomyocytes. Berberine exhibits variable positive and negative chronotropic effects in different species. Our first aim was to examine the effect of berberine in a cultured neonatal murine cardiomyocyte assay. Our study demonstrates that berberine has significant negative chronotropic actions on cardiomyocytes which is not an effect of beta-adrenergic receptor blockade. Pertussis toxin (PTX), a Gi/o protein inhibitor, blocked the negative chronotropic activity of berberine. Muscarinic, adenosine, opioid, and α₂ receptors are coupled through a G-protein (Gi/o) to adenylyl cyclase in an inhibitory fashion. Activation of these receptors are primarily responsible for PTX-sensitive negative chronotropic effects in heart. We hypothesized that berberine may be acting through one of these receptor type to decrease contraction rate of cardiomyocytes. For this purpose, we studied the effects of the muscarinic-receptor antagonists, atropine, himbacine, or AF- DX 116 on the negative chronotropic activity of berberine. Muscarinic antagonists completely blocked the effect of berberine on contraction rate of cardiomyocytes, whereas the bradycardic effect of berberine was not inhibited by the opioid, adenosine, or α2 receptor antagonists naloxone, CGS 15943, or phentolamine, respectively. Using [³H]QNB as a radioligand, we demonstrated that berberine bound to muscarinic receptors of adult mouse heart membranes with relatively high affinity. Furthermore, berberine dose-dependently inhibited [³H]QNB binding to muscarinic M2 receptors exogenously expressed in HEK 293 cells. Therefore, the findings of the present study suggest that berberine has muscarinic agonist effects in cultured neonatal murine cardiomyocytes, potentially explaining reported physiological effects of berberine. Cardiac hypertrophy represents the most important factor in the development of congestive heart failure. We investigated the inhibitory effect of berberine on hypertrophy of H9c2 cells. In rat heart-derived H9c2 myoblast cells treated with different hypertrophic agonists such as insulin growth factor II (IGF-II), arginine vasopressin (AVP), phenylephrine, and isoproterenol, protein content and size of cells were significantly increased compared to control group. However, the number of H9c2 cells after treatment with hypertrophic agonists did not differ significantly compared to control. The increases in area of cells and protein content induced by the hypertrophic agonists were inhibited by treatment with berberine in a concentration-dependent manner. Our findings have provided the first scientific evidence that berberine may have an inhibitory effect on hypertrophy of heart-derived cells, and provide a rationale for further studies to evaluate berberine's cardiac activity. / Graduation date: 2010