Analysis of Biochemical Compounds in ‘gynura Procumbens’

Analysis of Biochemical Compounds in gynura ProcumbensExtraction of Biochemical Compounds in gynura Procumbens Using 3 Mixture Design and GcmsName Syazliana Syamira Binti HashimNOVEMBER 2014genus genus genus genus genus Gynura procumbens or cognise locally in Malaysia as Sambung Nyawa or in chinese, it is called Nan Hui Ye is a whole kit and caboodle that was ground in South easterly Asia usually inThailand, Indonesia and Malaysia.This put belongs to the Asteraceae Family.It is a famous conventional herbaceous plant in South einsteinium Asia for the doment of diabetic, in high spirits cholesterol level, crabby person, high blood insistency, kidney discomfort and inflammation.The leave study aims to study the antidiabetic medicate application of Gynura procumbens leaves to manage diabetes and to study the authorization toxicity for many of the biochemical compounds in this plant leaves by using the three salmagundi designing and epitome by CGMS technique.In a mixtu re experiment, the independent factors argon proportions of different components of a blend. The fact that the properties of the different factors must sum to hundred% complicates the design as well as the summary of mixture experiments.In this work,a mixture design forget be applied to optimize solvent mixtures for selective extraction of biochemical compounds from Gynura procumbens leaves.Pressurized solvent extractions(PSE) will be performed at 313K and 20MPa,using homogeneous,ethanol(EtOH) and O solvent mixtures.The ope blackleging conditions to extract biochemical compounds from Gynura procumbens leaves will be optimized.The ranges of extraction temperatures and pressures ar 25 60 and 8 25MPa, respectively.GC coupled with MS detection will be utilise to characterize the extracts. baseThe modern lifestyle has become a major aff rectify to the wellness of people well-nigh the world. there are many cases that involve arteriosclerosis, obesity, diabetes mellitus and cancer s and these indispositions remind us about our unhealthy lifestyles and thus give rise in circulative system diseases such(prenominal) as high blood pressure, pre-hypertension and heart disease in positive countries (Johnson and Turner, 2005 Kearney et al., 2005).From the previous time,plants contri savee often been characterd to cure world diseases (Stepp and Moerman,2001 Yesilada, 2005).Today,with the arising threat of intractable diseases,a investigate is to focus on the finding bioactive molecules from plants(Amos et al.,2003 Fatehi et al.,2005).The one such plant is Sambung Nyawa or its scientific name is Gynura procumbens is a constituent of the Asteraceae family(Wiart,2002).Gynura procumbens is commonly utilize as a traditionalistic medicinal plant in Malaysia to treat many diseases. Recently, the use of herbal medicines for complementary discourses of some diseases has been popular and researchers constitute shown that these herbs are commonly used by cancer p atients to manage their diseases (Riboli and Norat, 2003 Van Duijnhoven et al., 2009).Pharmacologic studies in any case have proved that Gynura procumbens possesses anti-herpes simplex virus, (Nawawi et al.,1999), anti-ulcerogenic activities (Mahmood et al., 2010) anti-inflammatory drug (Iskander et al.,2002), and anticancer properties(Agustina et al.,2006).Problem account Gynura procumbens had been presumption attention in the pharmacology of antidiabetic medicinal plants probably because of its solid evidences and efficiency in the traditional vigilance of diabetes mellitus.But in contrast,the scientific reports on the antidiabetic activity of this plant had their own conflicts and inconsistent.For example,Zhang and topaz had report that 95% ethanol extract improved glucose tolerance in STZ-induced diabetic rats, but not in normal rats(Zhang and Tan).These authors excessively reported that its aqueous extract to exert significant anti hyperglycemic action in STZ-induced diab etic rats.But after that,Akowuah et al., on the contrary indicated its glucose lowering effect in normal rats(Akowuah et al.,2001).In the most recent study, the extract of Gynura procumbens was reported to picture significant elevation in the fasting blood glucose(FBG) levels of normal rats but a decrease in diabetic rats(Hassan et al.,2010).Eventhough Gynura procumbens is useful in the treatment of many ailments,however there is a little toxicological instruction that is available regarding to the safety due to the repeated exposure.Thus,a study need to be conducted to improve these reports and give more useful cultures about its widespread uses. therefore,the pose research is to study the antidiabetic activity of Gynura procumbens leaves to manage diabetes and also to study the say-so toxicity for many of the biochemical compounds in Gynura procumbens leaves.Objectives To study the antidiabetic activity of Gynura procumbens leaves to manage diabetes.To study the potential tox icity for many of the biochemical compounds in Gynura procumbens leaves.2.0 Literature reexamine2.1 General Overview of Gynura procumbensGynura procumbens that is well-known in South East Asia has traditionally been used to cure rash, eruptive fevers, kidney disease, constipation, migraines, diabetes mellitus, hypertension and cancer(Perry,1980).Recently, pharmacological studies have state that this plant has anti-hyperglycaemic(Akowuah et al.,2001 Akowuah et al.,2002),anti-hyperlipideaemic(Zhang and Tan,2000),anti-inflammatory(Iskander et al.,2002) and blood hypertension reduction capabilities(Lam et al.,1998 Kim et al.,2006).The advantages of the traditional use of Gynura procumbens leaves have been supported the isolation and identification of several manageable active chemical constituents from this herb, including flavonoids, tannins, saponins and terpenoids(Akowuah et al.,2002).Gynura procumbens is usually used as a traditional medicinal plant in Malaysia to cure many ailmen ts. It is an annual ever- putting green shrub with a fleshy stem and purple tint.In Malaysia,this plant is called as Sambung nyawa, and it is commonly used in South-East Asia,especially in Malaysia, Indonesia, and Thailand.In bitterness of the various uses over long time periods, little toxicological information is available regarding safety following repeated exposure to Gynura procumbens. Currently, Malayan authorities are paying more attention to the safety and potential toxicity of botanicals, including medical plants and edible materials. Therefore,the prey of the present study was to appropriate scientific data on the safety of Gynura procumbens, that focus on the acute and 90 long time (13weeks) sub-chronic toxicity of a wood alcohol extract that was orally administered to Sprague Dawley (SD) rats.Gynura procumbens has recently received particular attention in the pharmacology of antidiabetic medicinal plants,probably because of its admitted empirical evidence and effici ency in the traditional management of diabetes mellitus.Besides that, these study designs are not targeted at pictorial product uncovering or production of standardized herbal forms. Satisfactory research on medicinal plants beyond screening for biological activity should be conducted with the objective to systematically standardize and possess them into natural products or dosage forms which should effectively complement or supplement existing conventional measures(Ali et al.,2012).2.2 Current search of Gynura procumbensThere is presently a good deal of research interest to utilize plant compounds against human diseases, that includes hypertension. The present study investigated the make of different extracts and portions from leaves of Gynura procumbens on rat atrial concretion in vitro. Isolated left and right atria were mounted in a 20-ml pipe organ bath and they were allowed to equilibrate for 15 minutes before the application of the extracts or fractions. The extracts ( methanol extract (ME) and petroleum-ether extract (PE) ) and the fractions (chloroform fraction(CHL), ethyl-acetate fraction (EA), n-butanol fraction (NB) and water fraction (WA) of the methanol extract) were tested at three concentrations (0.25, 0.5 and 1.0 mg/ml), with a b-adrenergic protagonist (isoprenaline) as a control. exclusively the data on the contraction responses were log-transformed and had been analyzed. When overt to the different extracts, both atria tended to exhibit greater contractive responses with the NB whereas cardiac contractions had a tendency to be reduced with most new(prenominal) extracts. For a given extract, the contraction responses were particularly greater at 0.5 mg/ml for the right atrium and at 1 mg/ml for the left atrium. For the further analysis is to focus on the NB fraction revealed that positive inotropism was greater in left atria exposed to highly-concentrated F2 and F3 sub-fractions.Taken together,the results suggest that NB extracts a nd fractions from the Gynura procumbens-leaf methanol extract have positive inotropic activities and, hence, can be considered as an preference or traditional medicine against increase blood pressure in humans or can be used in strategies aimed at finding antihypertensive biomolecules from an accessible source.In each of these studies, the approach was not bioassay directed, an approach basis to the discovery of novel natural products from their natural sources and or to increase their standardization preparation to formulate into dosage forms for human usage after clinical trials. In summary, there is little analysis on the importance of the different extracts and fractions of Gynura procumbens on heart tissues. Therefore, the present study examined the effects of Gynura procumbens leaves extracts and fractions of the active extract on the contraction of left and right atria (LA and RA, respectively) isolated from Sprague Dawley (SD) rats with the aim of exploring the possibilitie s of its standardization or industrial scaling up for natural products against hypertension.2.3 Functions of Gynura procumbensIn South-East Asia, this plant is widely distributed and has often been used to treat diseases. In Indonesia, leaves of the Compositae family are routinely used for treatment in kidney diseases, eruptive fevers, rash, hypertension, diabetes mellitus, and hyperlipidemia(Perry,1980). In Thailand, where this plant is empirically used against topical inflammation, rheumatism, and viral ailments, Iskander and colleagues have reported the anti-inflammatory actions of its extracts(Iskander et al.,2002).Recently, pharmacological investigations in Singapore have shown that extracts of Gynura procumbens reduced blood serum cholesterol and triglyceride levels in streptozotocin induced diabetic rats(Zhang and Tan,2000). In Malaysia, evidence exists that Gynura procumbens has antidiabetic properties(Akowuah et al.,2001 Bohari et al.,2006). In this country, the currency o f hypertension is high, but the levels of awareness, control, and treatment are still low(Rampal et al.,2008).This species has been used as a traditional medicine to treat inflammation, cancer, rheumatism, and viral infections. Among the various ethnobotanical practices of this plant, the application as a poultice for several(a) skin diseases was found to be a universal usage in many countries (Perry, 1980). The leaves of this plant are not toxic(Rosidah et al., 2009). Gynura procumbensfrom Asteraceae family has been used as a traditional remedy for various skin diseases in certain areas of Southeast Asia(Junoh et al.,2011).2.4 Related DiseasesThere were many attempts to control change magnitude blood pressure that have been used as strategies to control arterial hypertension(Collins et al.,1990 Lang et al.,2001). Eventhough the treatment of arterial hypertension has been shown to be quite efficacious in reducing cardiovascular mortality rate and morbidity (Collins et al.,1990), hypertension control at the population level has been generally considered as not enough yet. In the United Kingdom, for instance, 94% of hypertensive patients still have their blood pressure higher than the normal(Colhoun et al.,1998), whereas in the United States, there is only 27% had a normal blood pressure following treatment(Burt et al.,1995). This intractability of hypertension has not changed, and recently there is a noticeable increase in hypertension cases(Kearney et al.,2004) about 26% of adults worldwide in 972 million are known to have hypertension(Kearney et al.,2005).Current optimism based on pharmacological therapy, which consists of developing drug formulations, is severely impeded by drug interactions, dose dependence, and adverse effects such as the possibility of depression. All in all,it is clear that the cultivation of alternative strategies complementary to existing control methods is needed.2.5 Acute and Sub-chronic ToxicityThe Gynura procumbens methanol ex tract was suspended in 1% (w/v) carboxylmethylcellulose (CMC) and it was administered orally (1250, 2500, and 5000 mg/kg) in single doses to both female and males rats (n = 10 5 males and 5 females). The rats need to fast overnight due to dosing. The general behaviour of the rat was continuously monitored for 1 hour after dosing, periodically during the first 24 hour (with special attention given during the first 4 hour), and periodical thereafter for a total of 14 days. All animals were humanely killed by inhalation after 14 days.The selected vital organs were excised, weighed, and macroscopically examined(Ghosh,1984). robust rats of both sexes were randomly assigned to control and treatment groups (n = 10 5 males and 5 females). The Gynura procumbens methanol extract was suspended in 1% CMC and administered orally on a daily basis for 90 days (13 weeks) at doses of 125, 250, or 500mg/kg. Based on the report by Suharmiati (2003), traditional healers typically prescribe seven leav es each day for an adult, which is equivalent to a 125 mg/day dose. Therefore, doses ranging from of 125500mg/(kg day) were selected as the tested doses. All rats were anesthetized under inhalation at the end of 13 weeks. Blood samples were amass via cardiac puncture into non-heparinized and EDTA containing tubes for biochemical and haematological analyses, respectively (Winarto, 2003 Petterino and Argentino-Storino, 2006). After cardiac puncture, the rats were killed by clavicle dislocation.Those vital organs were excised, weighed, and macroscopically examined and then they were meliorate in 10% formalin for histopathological study.2.6 Diabetes MellitusDiabetes causes aerobic stress through the autooxidation of glucose, protein glycation and lipid peroxidation and as a consequence, structural and functional alterations occur in the lungs of diabetics, as well as other organs like the heart, brain, eyes and kidneys. There is growing, well-established evidence regarding such chan ges in diabetic lungs(Gulay et al.,2014). Oxidative stress plays an grievous role in the pathogenesis of diabetes mellitus, contributing not only to the development, but also to the progression of diabetes and its related complications. The immunosuppressive and antioxidant effects of resveratrol in attenuating the increased oxidative stress due to responses of -cells to leukocyte activation have been implicated in the bar of type 1 diabetes mellitus. Resveratrol affords advantageous effects by decreasing the oxidative injury and the recruitment of the nutritive precapillary arterioles in the context of disease states associated with insulin resistance, such as metabolic syndrome, pre-diabetes, and type 2 diabetes mellitus.Globally, more than 366 million individuals currently live with diabetes mellitus, with this figure expected to rise to 552 million by 2030 (Whiting et al., 2011). The piece Health Organization estimates that around 1.1 million deaths occur per year as a result of the disease (World Health Organization, 2011). There are around 2.9 million individuals in England currently have diagnosed with diabetes, a currency of 4.5% (NHS The cultivation Centre, 2011). Projections for England estimate that by 2030 over 4.5 million people will be living with diabetes (APHO, 2011). The number of individuals that are living with diabetes means that if there was found to be an excess in mortality caused by the disease, this could result in a large number of premature deaths.The relation between diabetes and increased all-cause death, particularly from cardiovascular disease (CVD) and renal disease, is well recognized. C There is a limited amount of evidence relating to diabetes and respiratory disease mortality (Dawson et al., 2008De Marco et al., 1999andMurugan and Sharma, 2008).Current evidence suggests a biological link between inflammation, reduced levels of adiponectin inwardly the body and the development of diabetes and respiratory disease (Seveno aks Stockley, 2006). De Marco et al. concluded that these whitethorn be caused by differences in the severity of diabetes or differing in national treatment cultures (De Marco et al., 1999). Therefore, research is required that further explores the relation between diabetes and mortality from cancer and respiratory disease. Current research indicates that, for mortality from a number of causes, it may be diabetes-related comorbidities that increase an individuals risk of death rather than the diabetes itself. (Mohammadi et al., 2007) Morbidity and mortality of type 2 diabetes mellitus (DM) is closely related to development of cardiovascular disease (Haffner et al.,1998).Although the manifestations and complications of diabetes mellitus have been extensively studied, the knowledge regarding the effects of diabetes on bone mineral metabolism is still limited and inconsistent. Since Albright and Reifenstein(Anwar et al.,2008)first reported the occurrence of osteoporosis in patients wi th disadvantageously controlled diabetes in 1948, many authors have attempted to describe alterations of bone mineralization in diabetic subjects by using various techniques and biochemical markers(Asbun and Villareal,2006).The types of diabetes caused by other conditions or found in increased frequency with other conditions (imply an etiologie relationship) be considered a third subclass of diabetes mellitus that is the diabetes that associated with certain conditions and syndromes. This subclass has been divided according to the known or suspected etiologie relationships.The class gestational diabetes has been restricted to women in whom glucose intolerance develops or is discovered during pregnancy (National Diabetes Data Group,1979).