Phytochemical Analysis and hypoglycemic potential of Filago hurdwarica (Wall. ex DC.) Wagenitz in alloxan induced diabetic mice

Plants have profound therapeutic benefits, more economical treatments, fewer side effects, and a relatively cheap cost, making them a source of drugs for protective, preventative, curative, or conducive purposes and creating novel phytomedicines. Plant derived medicines are relatively safe compared to synthetic medicines. Many plants have proved to successfully aid in the treatment of diabetes including Filago hurdwarica (Wall. ex DC.) Wagenitz. The current investigations were therefore designed to assess the phytochemical, antioxidant, antidiabetic, and antihyperlipidemic activities of F. hurdwarica . The phytochemical investigations and antioxidant activities of different extracts were carried out using standard chemical tests, DPPH, and H 2 O 2 scavenging assays. F. hurdwarica plant extract in Hydromethanolic solution were prepared by Soxhletation method and stored in refrigerator at 4°C for two days before use. Swiss Albino mice were made diabetic by a single dose of alloxan (150 mg/kg). Hydromethanolic plant extract and fractions of F. hurdwarica were screened for antidiabetic activity and given to the alloxan-induced diabetic mice at a concentration of 150-250 mg/kg of body weight in different groups of 6 diabetic mice each orally once a day for 15 days. Glibenclamide is also given to another group to as a standard drug to support the result at a dose of 10 mg/kg of body weight orally once a day for 15 days. Blood glucose levels and body weights of mice were measured on 0, 4, 7, 11 and 15 th days. The study found that the extract was safe up to the dose level of 2000 mg/kg and the dose response effect of chloroform extract (150-250 mg/kg) of F. hurdwarica showed expressive antihyperglycemic effects and also improved other altered biochemical parameters associated with diabetes. The FTIR and XRD spectra demonstrated the occurrence of phenols, alcohols, alkenes, alkyl halides, ketones, and aromatic compounds and confirmed the amorphous nature of the extract. GC-MS spectral analysis showed the tentative presence of 31 phytochemical constituents in the chloroform extract of F. hurdwarica with different retention time. To conclude, the chloroform extract (250 mg/kg) of F. hurdwarica revealed considerable antioxidant, antihyperglycemic, and antihyperlipidemic potential and is safe for treating diabetes and related complications.


Ethics statement
All the experimental protocols involving animal experiments were developed in accordance with the approval of an ethical committee of the Department of Botany, Abdul Wali Khan University, Mardan.The animals were given feed ad libitum and 5% dextrose solution after 30 min of alloxan administration in order to stave off the early hypoglycemic phase.At the termination of the experiments, the animals were sacrificed by cervical dislocation under isoflurane anesthesia as per ethics committee guidelines, and all endeavors were made to reduce sufferings.

Collection of plant materials
The whole plant of F. hurdwarica was collected from District Buner, Khyber Pakhtunkhwa, Pakistan.The taxonomic identity of the desired plant was determined by the plant taxonomists at the Department of Botany, Abdul Wali Khan University Mardan, Pakistan.The sample specimen was deposited under a voucher no.Bot.20192 (AWK) at the herbarium of Botany Department (David et al., 2017).

Preparation of extracts
F. hurdwarica whole plant was collected, dried under shade, and then grossly powdered by a mechanical grinder.The powdered plant was weighed (1 Kg) and extracted with 80% methanol by direct maceration and allowed to stand for 3-4 days (David et al., 2017).The mixture was filtered after 72 h of maceration using a muslin cloth and then by Whatman filters paper No. 1.The filtrates were collected and the solvent was vaporized using a rotary evaporator (Heidolph Laborota 4000, Germany) at 40-50 °C.The dried extract was conveyed into a vial and preserved in a refrigerator until utilization (Mbiri et al., 2016).Solvent fractions were then prepared by liquidliquid separation method.Thirty grams of the crude

Introduction
Diabetes mellitus (DM) is a chronic multi-etiological metabolic disorder characterized by inadequate glucose homeostasis with a deficiency of protein, carbohydrate and fat metabolism disorders resulting from insulin action, insulin secretion, or both the defects (Akhtar et al., 2016;Fatima et al., 2019).DM has been characterized by hyperglycemia, hyperlipidemia, hypoinsulinemia, hyperaminoacidemia and oxidative stress due to a drop in insulin (Jyothi et al., 2012).Diabetes mellitus is known to cause hyperlipidemia because of the interrelationship between carbohydrates and lipid metabolism and through various metabolic derangements.Among several metabolic derangements, insulin deficiency has been known to stimulate lipolysis in the adipose tissue and give rise to hyperlipidemia.It involves Long-term complications of skin, kidneys, eyes, nerves, and blood vessels (Elosta et al., 2012).
Diabetes has turned up as a major health problem worldwide with serious health-related and socioeconomic impacts on individuals and communities alike.In both developed and developing countries, diabetes mellitus has affected about 25 percent of the global population (Benalla et al., 2010;Emdin et al., 2015).The multiple factors related to the development of diabetes are called risk factors or predisposing factors.The factors which increase the risk of diabetes include a sedentary lifestyle, high family aggregation, age, nutritional status, obesity, and insulin resistance (Deepashree and Prakash, 2007).
In both developed and developing countries, herbal medicine has gained growing attention in the treatment and management of diabetes, due to its origin and fewer side effects (Hasani-Ranjbar et al., 2009;Rahimi et al., 2005).The discovery of antidiabetic drugs has shifted its attention to medicinal plants to offer new potential and competent drugs with fewer adversarial effects and lower costs (Marles and Farnsworth, 1995;Nabi et al., 2013).In the development of novel therapeutic agents, natural products from plants play a major role and have gained great attention as sources of bioactive substances, including hypoglycemic, hypolipidemic, and antioxidant agents (Akhtar et al., 2016).The data from ethnobotanical studies reports about 1200 rare plants that may possess antidiabetic ability worldwide (Arumugam et al., 2013).
Brazilian Journal of Biology, 2024, vol.84, e261518 3/15 Antidiabetic activity of Filago hurdwarica methanolic extract was draped in 200 mL of distilled water in a separating funnel and it was then fractioned subsequently with 200 mL of various solvents of increasing polarity, commencing from n-hexane, followed by chloroform, and ethyl acetate, each in triplicate.The different solvent fractions were collected in flasks and then dried in an oven at 40°C (Tarekegne et al., 2016).The dried fractions were then transferred into vials and kept in refrigerator until used.

Quantitative determination of phytoconstitutents
The quantitative analysis of phytochemical constituents in the methanolic extract and fractions were carried out spectrophotometrically for the presence of phenol, flavonoid, tannin, sugar and protein contents by the "Folin Ciocalteu method" (Chun et al., 2003), Aluminum chloride assay (Chantiratikul et al., 2009), Folin Ciocalteu assay, Phenol Sulphuric acid method (Masuko et al., 2005) and Lowry's method (Ali and Sayeed, 1988) respectively.

Antioxidant study
The antioxidant potential of extract and various fractions of F. hurdwarica, was determined by using DPPH and Hydrogen peroxide assay methods (Ayoola et al., 2008).Ascorbic acid was used as a reference compound.

Experimental animals
The male Swiss albino mice of 3-5 weeks and weighing 25-30 g were used in the experiment.The animals were permitted to acclimatize in the animal house at the VIR (Veterinary Research Institute), Peshawar for a period of two weeks.The animals were kept at a narrowly sustained temperature of 25 ± 2 o C under optimum laboratory conditions.The experimental animals were used with the approval of an ethical committee of the Department of Pharmacy, Abdul Wali Khan University Mardan, (0012/2019).

Induction of diabetes
The diabetes was induced in Swiss albino mice by giving a single intraperitoneal (IP) injection of 150 mg/kg of alloxan monohydrate (Sigma Aldrich, Germany) (Ahmed et al., 2010).The animals were given feed ad libitum and 5% dextrose solution after 30 min of alloxan administration to overcome the early hypoglycemic crisis (Carvalho et al., 2003).The mice whose blood glucose level was more than 200 mg/dl were considered diabetic (Kesari et al., 2006).The treatment with the plant extracts was started after 3 days of alloxan injection in diabetic mice.

Acute toxicity test
The primary toxicological effect of the extract of F. hurdwarica was determined based on OECD guidelines 2008:425 (Sosa et al., 2020) to choose the suitable dose for antidiabetic assessments before commencement of the main study.The animals were observed for 96 hours to note the presence or absence of noxiousness and behavioral signs like tremor, restlessness, sluggishness, diarrhea, weight reduction and paralysis at regular intervals (Jörns et al., 1997).

Experimental design
The experimental animals were randomly divided into nine groups with six animals in each group for the evaluation of antidiabetic activity.Group 1, were normal saline-treated only with Tween 80, Group 2, were diabetic control received only alloxan 150 mg/kg, Group, 3 were diabetic mice treated with glibenclamide (10 mg/kg) (Ragavan and Krishnakumari, 2006), Group 4 and 5, had diabetic mice treated with crude methanolic extracts of 150 mg/kg and 250 mg/kg respectively, Group 6, 7, 8, and 9, included alloxan-induced diabetic mice provided with n-hexane, chloroform, ethyl acetate, and aqueous extracts respectively at a concentration of 250 mg/kg (Alema et al., 2020;Tafesse et al., 2017).They were marked separately and the treatment was continued for fifteen (15) consecutive days.

Determination of blood glucose
The blood glucose level was checked for each group on day 0, 4, 7, 11, and 15 during the study period by collecting blood from the mice tail vein of overnight fasted mice using a glucometer (Roche Diagnostics, Germany) and oxidase-peroxidase reactive strips (Morikawa et al., 2007).The tails were then rubbed with ethanol to prevent infection.The fluctuations in the weight of the animals were also recorded on the same days.

Determination of serum and lipid profile
The blood samples for serum profile were collected from overnight fasted mice under diethyl ether anesthesia by retro orbital plexus puncture method and were kept aside for 30 min for clotting.The serum was separated by centrifuging the sample at 3000 rounds per minute, for 10 min at 25°C (Morikawa et al., 2007).It was subsequently analyzed for alkaline phosphatase (ALP), serum creatinine, total cholesterol (TC), triglycerides (TG), high-density lipoproteins (HDL) and low-density lipoproteins (LDL) respectively by the methods (Friedewald et al., 1972).The SGOT (Serum Glutamic Oxaloacetic Transaminase) and SGPT (Serum Glutamate Pyruvate Transaminase) activities were determined according to the method of (Reitman and Frankel, 1957).These biochemical assays were measured by using commercial diagnostic kits.

Histopathological examination
The liver, pancreas and kidney of mice were cautiously dissected at the termination of the experiment and preserved in 10% formaldehyde.These were then cut into smaller pieces and fixed with 10% formalin solution and immediately processed for histopathological evaluation by parrafin method.The tissue segments were primed with microtome and stained with a hematoxylin and eosin dyes so as to be observed under a light microscope (Dwivedi and Daspaul, 2013).

In vitro membrane stabilizing activity
The membrane-stabilizing effects of the extracts were determined on human erythrocytes to predict the in vitro antidiabetic activity (Omale and Okafor, 2008).The percentage of hemolysis or membrane stabilization was calculated using the following Equation 1: where, optical density of test sample =

Measurement of antioxidant enzyme activity
Filago hurdwarica was investigated for its antioxidant potential in terms of antioxidant enzymatic capacities to support its traditional medicinal utilization.Enzymatic antioxidant activities were investigated by carrying out the Superoxide dismutase (SOD), peroxidase (POD, and Catalase (CAT) assays, spectrophotometrically at 25ºC by monitoring the absorbance at 420 nm, 560 nm, and 240 nm respectively (Onsa et al., 2004).

FTIR analysis
The FTIR (Fourier transform infrared) spectrum was used to detect the typical functional groups in the powdered extract of F. hurdwarica and was recorded from KBr pellet in FTIR Spectroscope (Shimadzu, IR Affinity1, Japan), with a scan range from 400 to 4000 cm -1 with a resolution of 4-1 cm (Pramila et al., 2012).

XRD analysis
XRD measurement was performed to validate the phase analysis of chloroform extract of F. hurdwarica according to the methods of (Li et al., 2012;Rajakumar et al., 2017).

GC-MS exploration of extract
The GC-MS analysis for the tentative identification of the compounds in the chloroform extract of F. hurdwarica was carried out by using GC-MS spectral analysis (Thermo Scientific Co.) (Hema et al., 2010).Interpretation of mass spectrum obtained from GC-MS analysis was done using the database of National Institute of Standard and Technology (NIST) library.

Statistical Analysis
The data was quantified as the Mean ± Standard Deviation (SD).The statistical investigations were accomplished using ANOVA followed by Dunnet post comparison test with the aid of IBM Statistical Package for Social Scientist (SPSS 20) software for data analysis.The variations were considered significant at P<0.05.

Quantitative determination of phytoconstitutents
The quantitative estimation of phytochemicals revealed that the plant extracts of F. hurdawarica contained phenols, flavonoids, tannins, total sugar, and protein as shown in Table 1.

In Vitro Antioxidant assay
The chloroform extract of Filago hurdwarica possesses the strongest ability to scavenge against DPPH radical (Table 2, Figure 1) and Hydrogen peroxide (Table 3, Figure 2) as compared to other extracts.

Acute toxicity
The extracts have been shown to be safe in mice up to the dose level of 2000 mg/kg body weight.There were no sensory and motor alterations, no changes in body weight, no toxicological effects, and no lethality detected in any mice.

Glucose tolerance test
Alloxan treated diabetic mice showed increased levels of blood glucose compared to normal mice (Table 4, Figure 3).After treatment with chloroform extract (250 mg/kg) of F. hurdwarica induced a significant hypoglycemic effect (p˂ 0.01) by restoring glucose levels to normal and its effect was comparable to that caused by standard hypoglycemic agent, glibenclamide used in the study.

Changes in body weight
In diabetic mice, the body weight was significantly decreased compared to normal mice.The body weight decline was significantly increased (p˂ 0.01) by the crude methanolic extract and chloroform fraction (250 mg/kg) at the end of 15 days of treatment.Other extracts were not effective (Table 5, Figure 4).

Antihyperlipidemic activity of plant
There was a significant elevation in the levels of serum cholesterol, triglycerides, LDL and reduced HDL in diabetic mice.The oral administration of chloroform extract (250 mg/kg) significantly reduced (p ˂ 0.01) the levels of TC, TG, and LDL and significantly elevated the levels of HDL (Table 6, Figure 5).

Serum profile
A significant decrease (p< 0.01) in serum creatinine and substantial increase (p<0.01) in SGPT, SGOT, and ALP   were observed in diabetic mice which showed renal and liver dysfunction caused by diabetogenic agent alloxan.The chloroform extract (250 mg/kg) of F. hurdwarica significantly improved the above parameters to about normal (Table 7, Figure 6).

Antioxidant enzymes
Diabetes mellitus significantly reduced antioxidant enzymes like catalases (CAT), peroxidases (POD), and superoxide dismutase (SOD) levels and elevated the action of reactive oxygen species.The chloroform extract at the dose of 250 mg/kg increased significantly increased the activities of antioxidant enzymes.However other extracts treated group showed no significant difference in comparison to the glibenclamide treated group (Table 8, Figure 7).

In vitro membrane stabilizing activity
In hypotonic solution induced condition, chloroform extract showed the highest protection of erythrocyte membrane (83.71%)which was comparable to haemolysis inhibited by the standard acetyl salicylic acid (Table 9).

XRD analysis
No distinctive diffraction peaks were observed in the whole spectrum, indicating the amorphous nature of chloroform extract of F. hurdwarica (Figure 9).

GC-MS analysis
The GC-MS analytical spectrum tentatively identified 31 bioactive compounds in the chloroform extract of F. hurdwarica with different peak area and retention time (Table 11, Figure 10).The efficient compounds stigmasterol and ß-sitosterol as depicted in Figure 11, identified in the GC-MS spectrum possess potential hypoglycemic and antioxidant properties.

Discussion
The current investigations were designed to evaluate the antidiabetic, antihyperlipidemic, and antioxidant potential of Filago hurdwarica in alloxan-induced diabetic mice with the view to validate its traditional utilization in the management of diabetes mellitus (DM) in humans.Alloxan generates diabetes in mice by specific toxicity effects, destroying pancreatic β cells and exhibiting the production of free radicals both in vivo and in vitro (Carvalho et al., 2003;Kamalakkannan and Prince, 2003;Lenzen, 2008).The fasting blood glucose levels above 200 mg/dl on the fourth day of administration of alloxan using glucose strips confirmed the induction of diabetes.The plasma glucose-lowering activity of the extracts was compared with glibenclamide, a standard hypoglycemic agent, used to cure diabetes mellitus for many years, by invigorating insulin secretion from pancreatic beta cells (Tian et al., 1998).
The oral administration of crude methanolic extract and the chloroform fraction of F. hurdwarica demonstrated a significant lowering of blood glucose level in the alloxaninduced diabetogenic animal model.The antihyperglycemic effect of F. hurdwarica may be due to the presence of active hypoglycemic agents or their ability to prevent free radical formation induced by alloxan.The reduction in blood glucose by the extract may also be attributed to the enhancement of peripheral glucose utilization in abdominal muscle and augmenting the pancreatic secretion of insulin from β-cells of islets of Langerhans (Jadhav et al., 2009;Singh, 2011).
In alloxan-induced diabetic mice, the reduction of body weight may be due to the breaking down of tissue protein and muscle wasting via unattainability of carbohydrates and catabolism of fats (Gougeon et al., 2008).However, treatments with orally administered methanolic, and chloroform extracts of 250 mg/kg significantly improved the body weight compared with diabetic control which signifies its protective effect in controlling muscle wasting i.e. reversal of gluconeogenesis, and subduing the free radicals produced due to hyperglycemia (Ma et al., 2017;Mestry et al., 2016).
In diabetes mellitus, hyperlipidemia occurs as a result of the excess mobilization of fats from the adipose tissue due to the underutilization of glucose (Akpan et al., 2012;Okon et al., 2007).Besides, hyperglycemia the study also demonstrated irregularities in the lipid metabolism, significant elevation in serum TC, TG, LDL, while reduction in serum HDL levels was observed in the diabetic mice before treatment.The glibenclamide (Group III) and chloroform fraction of F. hurdwarica (Group VII) treated mice showed a significant reduction in TC, TG, and LDL compared to diabetic control mice and a significant increase in HDL level.This could be due to increase utilization of glucose that led to lipid peroxidation inhibition and controlling of lypolytic hormones (Adeneye et al., 2011).A number of plants have been reported to possess anti-hyperlipidemic effects in this manner due to the presence of various phytochemicals in the plants extracts (Chaudhry et al., 2016;Yusufoglu et al., 2015).
It is acknowledged that the diabetes is linked to irreversible hepatocellular damage by destroying the liver    Antidiabetic activity of Filago hurdwarica microsomal cell, leading to the proliferation of different enzymes including SGOT, SGPT, and ALP into the blood (Pari and Latha, 2002).The crude methanolic extract and chloroform fraction (250 mg/kg) significantly lowered the levels of SGOT, SGPT, and ALP in the plasma compared to the diabetic control group.The diabetic mice showed a significant increase in serum creatinine level, which is considered as a marker of impaired renal dysfunction (Preethi and Kuttan, 2009).F. hurdwarica chloroform extract significantly reduced the serum creatinine thus treating and preventing the progression of renal damage in diabetic mice.Reports have shown that the reversal of such changes in above parameters is allied with the presence of secondary metabolites in the extract (Gupta and Gupta, 2009;Lal et al., 2009;Sharma et al., 2014Sharma et al., , 2019)).
In diabetes mellitus, high glucose can inactivate antioxidant enzymes SOD, POD and, CAT by glycating these proteins thus producing induced oxidative stress, which in turn, causes lipid peroxidation, which leads to both rise in ROS (reactive oxygen species) and to the demonetization of free radical scavenging compounds (Baynes and Thorpe, 1999;Kakkar et al., 1998;Karasu, 1999).The extensive array of antioxidant enzymes, SOD, POD, and CAT shield the body from the hostile effects of free radicals that are produced in vivo under normal physiological circumstances (Halliwell, 1999).SOD scavenges superoxide anion to form hydrogen peroxide reducing the toxic effects derived from a secondary reaction.POD and CAT is involved in the reduction of hydrogen peroxide thus protecting the erythrocytes from reactive oxygen species (Karasu, 1999).Treatment with methanolic and chloroform extract has reversed the activities of these enzymatic antioxidants which could be a result of decreased lipid peroxidation and decreased utilization (Yin et al., 2014).
Several studies revealed that injury to membranes of RBCs due to hyperglycemia makes cells more susceptible to secondary damage through lipid peroxidation, haemolysis, and oxidation of haemoglobin (Halliwell, 1999).The chloroform extract of F. hurdwarica showed significant inhibition of hypotonic solutioninduced hemolysis of erythrocytes compared to control.The inhibition of erythrocyte lysis property of F. hurdwarica could be the possible mechanism for its anti-diabetic activity due to the presence of various biochemical compounds in the extract which exert a profound stabilizing effect on lysosomal membrane and their cations binding ability (Miliauskas et al., 2004;Oyedapo et al., 2004).
Various secondary metabolites isolated from diverse plant species have been shown to possess potent antihyperglycemic and glucose suppressive effects.These secondary metabolites include alkaloids, flavonoids, terpenoids, and phenols (Ivorra et al., 1989;Sharma et al., 2010).These might stimulate glycogenesis in the liver, insulin release from pancreatic ß-cells, or inhibiting glucose absorption in the gut, or increasing glucose utilization by the body (Sezik et al., 2005).As a result, these secondary metabolites which were found in F. hurdwarica crude extracts and chloroform fraction may, therefore, be accountable for the observed glucose suppressive and anti-hyperglycemic activity (Kumar et al., 2011).The antioxidant effect of the extract may involve the scavenging of free radicals as seen with DPPH and H 2 O 2, thereby protecting the cells from the damaging effect of the free radicals and successive complications.Antioxidants have been recognized to exhibit protective functions against oxidative stress and are associated with reduced risk of chronic complications (Gordon, 1990).The experimental finding revealed that the chloroform extract has significantly high DPPH and hydrogen peroxide radical scavenging activity which is an agreement with the previous studies (Ali and Sayeed, 1988;Asamenew et al., 2011).
The characteristic peaks of different functional groups from the FTIR spectroscopic characterization show the presence of relative active compounds in the sample that might be used as antidiabetic and antihyperlipidemic agents as reported in previous studies (Sidhu and Sharma, 2014;Wei et al., 2017).XRD analysis of F. hurdwarica showed no distinctive diffraction peaks in the whole spectrum suggesting the amorphous nature of the extract.However, the possible presence of amorphous phases could be attributed to the phenols, alkaloids, flavonoids, and tannins, which are evident from the qualitative and quantitative screening tests.Furthermore, the result of XRD confirmed the conclusion of FTIR analysis.These results are in accordance with previous studies suggesting the amorphous nature to the presence of phytoconstitutents (Bala et al., 2015;Rajaram et al., 2015;Varadharaj et al., 2019).The GC-MS spectral data tentatively identified that the chloroform extract contained several compounds that were found active in terms of antidiabetic and antioxidant potentials as evident from their reported literature.In diabetic context stigmasterol and ß-sitosterol exhibit antidiabetic and membrane-stabilizing effects (Nualkaew et al., 2015;Ramu et al., 2016;Zeb et al., 2017).Similarly, amiodarone, hexadecanoic acid, and 2,3-dihydroxypropyl ester are also used as hypoglycemic agents either alone or in combination (Lai et al., 2020).Other antidiabetic compounds identified included, 1-Monolinoleoylglycerol trimethylsilyl ether (Senthil et al., 2016), vitamin E (Bharti et al., 2012), and Dasycarpidan-1-methanol, acetate (Al-Gara et al., 2019).

Conclusion
The current research study revealed for the first time the antidiabetic and antioxidant potential of F. hurdwarica in alloxan-induced diabetic model.The study demonstrated that the chloroform extract of F. hurdwarica possess substantial antihyperglycemic, antihyperlipidemic, and antioxidant potential in alloxaninduced diabetic mice models thus extenuating its traditional utilization.The preliminary information obtained from the FTIR, XRD, and GC-MS procedures recommended the therapeutic significance of the plant.Consequently, further study is indispensable for hypoglycemic mechanism and isolation of bioactive constituents for auxiliary validation of the plant.Future research needs to focus on bioassay guided isolation and confirmed identification of the active principles using standards or spectral workup.This could inform better extraction and formulation procedures to lower the dose required, as well as lead to a validated phytochemical analysis of a standardized product.

Figure 3 .
Figure 3. Changes in blood glucose level of mice.Figure 4. Changes in body weight of mice.

Figure 4 .
Figure 3. Changes in blood glucose level of mice.Figure 4. Changes in body weight of mice.

Figure 7 .
Figure 7. Antioxidant enzymes activity of plant extracts.
Data presented as mean ± SEM of each triplicate test.

Table 4 .
Effect of extracts of F. hurdwarica on blood glucose.

Table 5 .
Effect of extracts of F. hurdwarica on body weight Values are expressed as mean ±SEM n=6 in each group.*P < 0.05; **P < 0.01, significant results compared with diabetic control.

Table 6 .
Effect of extracts of F. hurdwarica on lipid profile.

Table 7 .
Effect of extracts on serum profile.

Table 8 .
Effect of F. hurdwarica extracts on antioxidant enzymes.

Table 9 .
Effect of plant extracts on RBCs membrane stabilization.
Data are expressed as mean ± SD (n=3).