A and RNA contents and also nucleases (RNAase and DNAase) activity. The material and Methods:The study was performed in six groups. The first group was the normal healthy control; the second group was rats injected with alloxan (diabetic control).
Every one of the other four groups (diabetic rats) was ingested with each of the methanolic extracts of Kafta, Doum, Sammo and Araar (individually).
The results: The results of chromosomal analysis showed that, diabetic rats had a highly significant increase of chromosomal aberrations compared to that of normal healthy control.
Animals which treated with the four plants methanolic extracts showed significant improvements in the frequencies of chromosomal aberrations. In case of biochemical analysis, blood glucose level was significantly increased but immunoglobulins levels were decreased in diabetic albino rats. Diabetes increased serum total lipids, cholesterol, triglycerides and LDL but serum HDL was decreased. RNA and DNA as well as inhibited the nucleases (RNAase and DNAase) activity of both organs (liver and testes) tissue. The conclusions: It is concluded that diabetes is much harmful in the animal body, whilst the induction of flavonoids extract (by methanol) of Kafta, Somma, Araar and Doum reduced these harmful of diabetes. Depending on the severity of the metabolic abnormality diabetes may asymptomatic, or may be associated to symptoms (thirst polyuria and weight loss) or may be progress to ketoacidosis and coma [2].
It is associated with both acute and chronic complecation, chronic hyperglycemia cause damage to the eye, kidneys, liver, nerves, heart and blood vessels [3]. Flavonoids are among the compounds that commonly occur in plants and are frequent components of the human diet.
The antioxidative characters of these compounds was reported before in several studies [4,5]. Diabetes related to oxidative stress has potentially serious consequences and limited treatment research has focused on arresting the oxidative damage that may be involved in disease initiation or progression. MATERIALS AND METHODS 2.1. Chemicals Alloxan: recrystallized, used to produce insulin – dependent diabetes experimentally. Plant Matrials Methanolic extracts of Anastatica hierochuntica (Kafta) Hyphaene thebaica (Doum) Cleome droserfolia (Sammo) Juniperus phoenicea (Araar) are used as antidiabetic plants. Animals Adult male rats (36 rats) (Rattus norvegicus) weighted 100-120 gm obtained from a close random bread colony at National Research Center. The animals had free access to standard pelleted diet and tap water.
Standard diet consists of casein 15%, cotton seed oil 10%, salt mixture 4%, vitamins mixture 1%, starch 65% and cellulose 5% [7]. Experimental Design Adult male rats were divided equally into six groups: 1) Normal group (control healthy group) received only distilled water. Chromosomal Analysis The male rats were sacrificed 24h. Biochemical Studies At the end of the experimental period, animals were killed by decapitation. Blood, liver and testes of the experimental animals were collected. Liver and testes chilled up for analysis and blood serum was prepared by centrifugation at 3000 rpm and chilled up for analysis. Total soluble protein of both organs tissue was carried out according to Lawary et al [16] method. DNAase and RNAase activities of liver and testes tissues were dete rmined according to the methods described byBergmeyer [17].
Statistical Analysis Data for chromosomal analysis were analyzed using Chi-square test [18]. A statistical significance between the treated and control groups was done.
Somatic Cells (Bone Marrow Cells) The chromosomal examination in this study showed that the structural aberrations include gaps, breaks, fragments, deletions and endomitosis.
Numerical aberrations involved polyploidy, hyperploidy (included cells with 41 and 43 chromosomes) and hypoploidy (included 38 and 39 chromosomes) were also studied and showed differences from that of the normal control group (Table 1), in the four tested plants and Alloxan group (diabetic rats). Spermatogonia The chromosomal examination of spermatogonial cells in diabetic animals after and before treatment with plants showed more frequencies of structural and numerical aberrations of chromosomes than the normal group (Table 2). Spermatocytes Table 3 presents the results of structural and numerical abnormalities in spermatocytes of all studied groups.
The observed types of abnormalities were x-y univalent, autosomal univalent, rings and chain. Diabetic rats (Alloxanized) showed a highly significant increase at level (p Table 3) and showed a highly significant increase (P Cytogenetic studies showed that Araar and Doum had the lowest percentages of total structural aberrations, but generally statistical analysis showed that there were no significant differences between the four medicinal plants used. Mean percentages ±SE of chromosomal aberrations in rat bone marrow cells after treatment with Kafta, Doum, Sammo and Araar. Mean percentages ±SE of chromosomal aberrations in rat spermatogonial cells after treatment with Kafta, Doum, Sammo and Araar. Mean percentages of chromosomal aberrations in rat spermatocytes after treatment with Kafta, Doum, Sammo and Araar. These results showed that diabetes significantly decreased total soluble protein to 67% and 73% at normal control, respectively for liver and testes.
The ingestion of methanolic extracts of the present studied plants improved these abnormal contents which were 101-102% and 107-110% at normal control for liver and testes tissues, respectively of diabetic rats.
Influences of Kafta, Doum, Somma and Araar methanolic extracts on blood glucose and total soluble protein of liver and testes tissues of diabetic male adult albino rats.
Influences of Kafta, Doum, Somma and Araar methanolic extracts on DNA and RNA contents of liver and testes tissues of diabetic male adult albino rats. In contrast, the administration of methanolic extracts of the four medicinal plants (Flavonoids) increased and improved the reduced values of RNA and DNA of diabetic animals which readjusted to those of normal control either in liver or testes which were 101-117% at those of normal healthy control rats.
Consequently, the activity of RNAase and DNAase (nuclease enzymes related to nucleic acids system) were determined in liver and testes tissue of the experimental animals. The obtained results are shown in Table 6" target="_self"> Table 6, which observed that significant inhibitions were found in the DNAase and RNAase activity of the both organs tissue of diabetic rats related to healthy control. The inhibited activity by diabetes was ranged between 50% and 61% at normal control for the two nucleases activity (DNAase and RNAase). This may be due to the oxidative stress in diabetes coexists with reduction in the antioxidant defense system and subsequently leads to the complications of diabetes [38]. Results of Table 6 clearly showed that significant stimulation were caused in the activity of both nucleases of liver and testes tissues of the diabetic rats by ingestion of the methanolic extracts of Kafta, Somma, Doum and Araar. The activity values were improved around those of the normal healthy control in which the DNAase activity was 101-103% at control in liver and testes tissues of diabetic rats ingested with each of the four methanolic extracts but for RNAase the activity was 102% in liver and 107-112% in testes tissues of diabetic rats relative to normal healthy control by the same treatments.
Diabetic rats ingested Sammo extract gave the highest levels of the three immunoglobulins when compared with the other used three medicinal plants.
The results observed that total lipid, LDL-C and total triglycerides (levels of serum of diabetic rats) amounted about 2 fold, while the total cholesterol of serum diabetic rats content was 1.5 fold than that of normal healthy control but HDL-C level of diabetic rats Table 6.
Influences of Kafta, Doum, Somma and Araar methanolic extracts on DNAase and RNAase activity of liver and testes tissues of diabetic male adult albino rats.
Data in Table 8 found that all the used plants methanolic extracts significantly improved the lipid profile of diabetic rats serum in which total lipids, triglycerides, cholesterol and LDL-C contents in serum of the diabetic rats were significantly decreased and the values of ingested diabetic rats were around that of the healthy normal control, but HDL-C level of serum diabetic rats was significantly increased and their levels were around or more than that of healthy ones. The ingestion of Sammo methanolic extracts into diabetic rats produced the highest effects on lipids profile of diabetic rats serum. DISCUSSION Diabetes mellitus is one of the most common metabolic diseases. In many countries the incidence of diabetes mellitus has reached a high percentage. In a survey many of non-insulin-dependent diabetes was found to resort to the use of medicinal plants to treat their diseases [20]. In fact, phytotherpay has been widely used because of the low cost and the easy availability of medicinal plants. However, the studies on antidiabetic plants used are relatively recent and has begun to evolve in the last few years. Moreover, until recently, there has been no extensive documentation of data on antidiabetic plants.
Alloxan produce insulin-dependent diabetes animals experimentally. It reduced the number of dividing cells causing inhibition in the mitotic index than normal. Alloxan and streptozotocin (STZ) can destroy ?-cells causing significant increase in blood glucose level. This increase in circulating glucose level is believed to contribute to ?-cell dysfunction [21].
This effect relates to their structure as a DNA alkylating agents, w.hich leads to necrosis of pancreatic beta cells and thus to a state of insulin-dependent diabetes mellitus [22]. The action of these drugs could also be due to alternation in membrane fatty acid content, which may affect Na+, K+ - ATPase activity, membrane fluidity, and fatty acid content [23]. Oral treatment with methanolic extracts of Kafta, Doum, Sammo and Araar inhibited the frequency of chromosomal aberrations significantly in both somatic and germ cells.
It showed a hypoglycemic effect on blood glucose level in these animals. They reported that diabetes caused an oxidative stress include stimulation lipid peroxidation and protein glycation as well as inhibition of antioxidative enzymes with alteration in the structure and function of collagen and basement membranes as well as reduced tissue soluble protein. Natural flavonoids such as those of Kafta, Somma, Araar and Doum methanolic extracts have been shown to reduce this oxidative stress in experiemental diabetes [31, 32]. The lowering effect of plant flavonoids on diabetes may be due to the increase in insulin-receptor binding followed by increasing the signal sequences [33] or the antioxidant of these flavonoids.
The oxidative stress usually results in stimulated superoxide dismutase activity and inhibited insulin binding to insulin receptor [34]. These effects of diabetes were improved by flavonoids treatment (presented in the methanolic extracts of the studied medicinal plants, [35]. The improved effects of methanolic extracts on diabetic rats are in agreement with those of Halima – Abdou et al., [29] who found that in pregnant rats, flavonoids extracts improved the injurious effects of diabetes on the activities of nucleases. The stimulation of nucleases activity of diabetic rat tissues under the treatments of flavonoids of plant methanolic extracts may be due to that diabetes caused in animal an oxidative stress which produced great chromosomal aberrations. These damaged DNA and blocked the action of RNA polymerase and so prevent replication process.
The resulting damage must be repaired under the induction of flavonoid treatment if DNA is return to its genetic integrity. The DNA damage is excised by the sequential of nuclease leaving a short gap which is filled in by action of polymerase.
Recent studies have revealed that adiponectin plays important roles in the regulation of lipid metabolism. For example, it increases free fatty acid oxidation in the muscle, enhances hepatic insulin action and inhibits foam cell formation by reducing the accumulation of lipids in human macrophages and prevents atherosclerosis. These may be due to stimulation of protein biosynthesis process to produce the specific factors (included antioxidative enzymes) required to improve and treat the oxidative stress of diabetes by replication and transcription needs a lot amount of nucleotides, which obtained in the tissue cells by hydrolysis of DNA and RNA via nucleases [39]. Press Inc., New York, San Francisco and London, 447-511.




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