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The influences of medicinal plants Juniperus Phoenicea (Araar), Hyphaene thebaica (Doum), Anastatica hierochuntica (Kafta) and Cleome droserifolia (Sammo) as antidiabetic agents were investigated using female pregnant albino rats. Female rats were injected with 60 mg/kg b.w. alloxan to induce diabetes. Diabetic rats treated orally with the methanol extracts of tested plants till the 19 day of gestation. The present studies include the frequencies of chromosomal aberrations and nucleic acid system of liver in the female pregnant rats and their embryos. The results showed that injection of alloxan caused highly significant increase in chromosomal aberrations as well as in blood glucose levels as a result of diabetes in pregnant females. It also caused a high incidence of chromosomal deviation in embryos and decreased the liver soluble protein contents of female rats and their embryos. These effects in alloxanized animals were treated and improved by ingestion of the methanol extracts of the tested plants (Araar, Doum, Kafta and Somma) in which under their treatments, the inceased level of blood glucose of diabetic rats was deceased. Ingestion with the plants methanolic extracts improved and normalized the effects of diabetes in nucleic acids values of liver tissues. These were accompanied with nucleases (RNAase and DNAase) activities. The inhibited activities of both DNA ase and RNA ase of pregnant rats and their embryos were stimulated and readjusted around the normal values. Also administration of the plants methanol extracts decreased the percentage of chromosomal aberrations in the female rats and embryos. It is concluded that there are some biochemical dynamics which might occur in the metabolism of glucose, nucleic acids and proteins in order to prevent or to reduce the oxidative stress of diabetes by flavonoides treatment.
Diabetes mellitus has become one of the most frequent complications â accompanying chronic diseases with high morbidity and mortality (Adeghate, 1999 b). It is a metabolic disorder characterized by hyperglycemia resulting from lack of insulin, lack of insulin effect, or both. Two types of diabetes are recognized; type 1, known as insulin â dependent diabetes (IDDM), and type II, known as non â insulin â dependent diabetes (NIDDM) (Hansen, 1998). In type I diabetes, the absolute deficiency of insulin attributed to βâcell dysfunction leads to hyperglycemia, enhanced lipolysis, protein catabolism and ketosis (Hansen, 1998). Type II diabetes impairs the ability of insulin to stimulate both muscle glucose uptake and splanchnic glucose uptake, thus contributes to induce hyperglycemia (Basu et al., 2000). The severity and frequency of the late degenerative complications are high in patients with either type I or type II diabetes mellitus (Adeghate, 1999 a). The available methods for treatment of diabetes mellitus include diet or diet and oral hypoglycemic drugs or diet and insulin (Boon et al., 1999). In traditional practices, medicinal plants are used to control diabetes mellitus in many countries. This caused an increase in the number of experimental and clinical investigations directed toward the validation of the hypoglycemic (ElâRidi, 2001) and / or anti â diabetic (AlarconâAguilara et al., 1998) properties of different medicinal plants. It has been reported that the chard (Beta Vulgaris L.var. cicla) extract increased pancreatic beta cells in Streptozotocin (STZ) â diabetic rats (Bolkent et al., 2000). Water extract of smallantus Sonchifolius (yacon) leaves fed to normal and STZ diabetic rats also showed hypoglycemic effects (Aybar et al., 2001). Moreover, a long â term administration of olive leaves or Cinnamon bark caused significant improvement in tissue injury induced by STZ treatment (Onderoglu et al., 1999). The water extract of Anastatica hierochuntica, in the dose given and rout of administration used by Tarek and Mamdouh (2002) has hypoglycemic effect in both normoglycemic and diabetic rats. The objectives of the present study are to evaluate the hypoglycemic effect of the methanolic extracts of J. phoencea, H. thebaica, A. hierochuntica and Cleome droserifolia in normal and Alloxan â diabetic pregnant female rats and their embryos.
2.1.Animals Adult diabetic and normal albino rats (72 rats) weighing (120 â 150 g) were maintained in a well ventilated animal house. Animals were housed in large spacious polypropylene cages with free access to food and ad libitum during the course of experiment (Lane â patter and Pearson, 1971). Males and diabetic females were orally administered with the different treatments once daily, for one week, then two females were housed with one male overnight, and successful mating was confirmed by vaginal plug. The presence of vaginal plug was considered as the zero gestational day. The pregnant female's administration with the different treatments continuous during pregnancy till the 19th day of gestation. 2.2. Chemicals: Alloxan: recrystallized, used to produce insulin â dependent diabetes experimentally. 2.3. Plant matrials: The methanol extracts of the four tested plants: C. droserfolia (sommo), J. Phoenicia (araar), A. hierochuntica (kafta) and H. thebaica (doum) were used in the present studies as antidiabetic agents. 2.4.  Experiments Pregnant females albino rats were divided into six groups (36 rats) Group I control group (normal pregnant females) was orally administered with distilled water,Group 2 animals were injected with alloxan 60 mg/kg (Lazaro, 1949) for only one injection and orally administered with distilled water (pregnant diabetic control), Group 3     pregnant diabetic rats were administered orally with 150 mg/kg/day with methanolic extract of sammo,Group 4p pregnant diabetic rats administered orally with 150 mg/kg/day with methanolic extract of kafta,group 5 pregnant diabetic rats administered orally with 150 mg/kg/day with methanolic extract of araar and group 6    pregnant diabetic rats administered orally with 150 mg/kg/day with methanolic extract of doum. Another six groups (36 rats) of nonpregnant females albino rats were used as controls for previous six groups, respectively for blood glucose values comparison. Females were sacrificed at day 19th of gestation and the chromosomes prepared from bone marrow cells of mothers according to Yosida et al. (1977). In case of embryo, cells were prepared according to the method of Romagnano et al. (1985) to study any genotoxic effect. For biochemical studies blood and livers of the females and their embryos were collected. Livers chilled up for analysis and blood plasma was prepared by centrifugation at 3000 rpm and chilled up for analysis (Astrwov, 1974). 2.5. Slide preparation: To study the chromosomal aberrations in bone marrow cells of females (mothers), 50 metaphases from each animal were examined. In the case of embryo cells, chromosomal aberrations of liver in 900 cells per each group were examined. Determination of blood glucose was adopted using the method of Trinder (1969). Liver tissues were homogenized by the method of Peares (1985) for determination of RNA and DNA. The DNA concentrations were determined in the supernatant according to Dische method (1955) using diphenylamine reaction which produced blue color when heated with DNA. The RNA concentrations were determined using oricinol method according to Schneider method (1957). Total soluble protein in liver was determined according to Lowary et al. (1951) method. The activity of liver nucleases was determined according to Bergmeger (1974). 2.6. Date analysis Intergroup differences of cytogenetic and biochemical studies were analyzed statistically by using standard T-test according to Sokall and Rohlf (1995).
3.1. Cytogenetic results: The present result in Table (1) showed the frequencies of structural and numerical chromosomal aberrations as well as the mitotic activity induced by alloxan and also the effect of administration of the methanol extracts of the plants studied in female bone marrow cells. Injection with alloxan increased the frequencies of individual and total chromosomal aberrations in female bone marrow cells when compared with the control. This increase was found to be statistically significant at (P
The increase in number of diabetic patients have motivated scientists to find new method to cure diabetes (Adeghate, 1999 b). In the present study, the effect of the metabolic extracts of J. phoencea (Araar), H.thebaica (Doum), A. hierochuntica (kafta) and C. droserifolia (Sammo) were tested on normoglycemic as well as alloxan diabetic female pregnant rats and their embryos. The diabetogenic effect of alloxan and STZ was reported by several investigators (Choi et al. 1991; Adeghate, 1999 b; Wang et al., 2000; Mohamed Bnouham et al., 2002). It was indicated in the present study by the significant increase in blood glucose level in alloxan injected rats. It was also indicated by the highly significant increase in somatic chromosomal aberrations of pregnant female and their embryos. The effect of STZ and alloxan relate to their structure as a DNA alkylating agent, which leads to necrosis of pancreatic beta cells and thus to a state of insulin - dependent diabetes mellitus (Elsner et al., 2000). As alloxanized diabetes may either increase the entevance rate of glucase into blood from liver (stimulated hepatic glycogenolysis or gluconeogenesis) and may be reduced the rate of removal of glucose from blood by different tissues which decreased storage or utilization of glucose. These may be due to the absent of insulin. Alloxan acted directly promptly and specifically on the βâcells of pancreas (Chatterjea and Shinde, 2002). On the other hand, the results indicated that administration of the methanol extracts of the present studied plants improved, but not completely normalized, the diabetogenic action induced by alloxan. They showed hypoglycemic effects on blood glucose level in plant â treated diabetic females and embryos when compared with untreated diabetics. They also caused a significant decrease in the percentage of chromosomal aberrations caused by alloxan in both mothers and embryos. The four methanolic extracts had a great amount of flavonoid compounds which did not alter the low level of insulin (Ismail et al., 2007). This means that the reducing effect of methanol extracts (flovonoid compunds) on blood glucose of diabetic animals was not related to the insulin content in blood. Flavonoids ingestien into diabetic animals increased blood adiponectin level, which produced a hypoglycemic action of insulin sensitivity (Comds et al., 2002; Yamauchi et al., 2003; Ismail et al., 2007). Similarly, it is believed that the presence of flavone glycosidic components in the plant extract of Anastatia hierochuntia (Khalifa, 1980), is responsible for reducing the blood glucose level in STZ â diabetic rats. Such hypoglycemic effect could be through increased serum insulin levels provided by repair/ regeneration of the endocrine pancreas. Results of liver total soluble protein nucleic acids and nucleases showed that diabetes caused a great harmful and disturbance in liver metabolism including either in liver metabolites content or in enzymes activity, but the medicinal plants methanol extracts (included falvonoids) improved and normalized these harmful and disturbed effects of diabetes on livers metabolism as well as their metabolites. The present results are in agreement with those of Salah et al. (2010). They found that flovonoids of the metabolic extracts of Kafta, Araar, Somma and Doum improved and treated diabetes in case of blood glucose, and liver contents of total soluble protein, RNA and DNA as well as the activity of enzymes related to nucleic acid such as nucleases (RNAase and DNAase). They also found that it inhibited the percentage of chromosomal aberrations in male somatic and germ cells. It means that, the effects of diabetes oxidative stress on liver protein and nucleic acids as well as their nucleases activity was readjusted and improved by ingestion of alcoholic extracts of the four present medicinal plants. On the other hand, there is no any available data was observed in literature about these items in pregnant animals and their embryo, except those of our work on male albino rats. The present findings of DNA content in liver of pregnant females and their embryos are confirmed by the results of chromosomal abberrations in which DNA content was deceased and chromosomal aberrations were increased by diabetes. Also, RNA content data confirmed the results of protein content which is in agreement with the findings of nucleases activity. This means that the present results confirmed each other. The results showed that plants methanol extracts (including flavonoides) might cause an adaptation of certain enzymes system in the animal tissues of diabetic animals which provide the necessary enzymatic mechinary to cope with the increased flow of nucleotides during the experimental period by converting it rapidly into DNA and RNA (m-RNA, r-RNA and t-RNA). However, flavonoides may play a role in the improvement of replication and transcription enzymes activity to produce DNA and RNA. In addition, the efficiency of flavonoides against the oxidative stress of diabetes is not completely known (Coskun et al., 2005 and Ismail et al., 2007). Flavonoides were in conjugation with adiponectin may play a role in binding of insulin with its receptor. Moreover, these compounds may interfere with signal sequences of insulin inside the cell (Anton et al., 2007 and Abdel-Magid, 2007). The induction of flavonoides extracts elevated the adiponectin levels in the diabetic rats, which showed glucose lowering effects and improvement in insulin resistance and sensitivity and diabetic oxidative stress as well as the whole body metabolism (Abdel-Magid, 2007) From the present results, it can be concluded that there are some biochemical dynamics which might occur in the metabolism of glucose, nucleic acids and proteins in order to prevent or to reduce the oxidative stress of diabetes by flavonoides treatment. The decrease of blood glucose with increase soluble protein and nucleic acids content as well as stimulation of nucleases activity of liver tissues in diabetic female pregnant rats and their embryos by flavonoides induction subsequently is considered as results of diabetes oxidative stress which was treated by flavonoides compounds. Also, the results suggested that, the stimulation of protein biosynthesis processes was marked by increase the RNA content as well as stimulation of nucleases activity. The enzymes may supply transcription stage of protein biosynthesis with the needed precursors nucleotides (Adams et al., 1993).
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