Evaluation of Antidiabetic and Anti- Hyperlipidemic Activity of Flower lantana Camara Linn Extract in Streptozotocin-Induced Diabetic Rats Model

Diabetes mellitus is a chronic disorder of carbohydrate, fat, and protein metabolism, mainly due to insulin deficiency or resistance [1] STZ-induced diabetes in rats mimics human Type 1 diabetes and is commonly used in preclinical studies. Hyperglycemia often leads to hyperlipidemia, increasing the risk of cardiovascular complication [2] Lipids - Diabetes mellitus produces abnormal changes in the lipid profile. It will lead the cells to become more susceptible to lipid peroxidation [3]. Experimental studies show that the presence of polyunsaturated fatty acids in cell membrane leads to attack by free radicals due to the presence of multiple bonds [4]. Lipid hyper peroxides (LHP) through intermediate radical reactions produce such fatty acids that generate highly reactive and toxic lipid radicals. They form new lipid hyperperoxides [5]. A critical biomarker of oxidative stress is lipid per oxidation. It is the most explored area of research when it comes to ROS [6] Malondialdehyde (MDA) is formed as a result of lipid peroxidation that can be used to measure lipid peroxides after reaction with thiobarbituric acid. These conditions lead to increased levels of markers of oxidative stress and dyslipidemia Theories behind elevationin liver function parameters in diabetes state that the liver helps to maintain normal blood glucose concentration during fasting and postprandial states. Loss of insulin effect on the liver leads to glycogenolysis and increase in production of hepatic glucose. Abnormalities of triglyceride storage and lipolysis in insulin-sensitive tissues such as the liver are an early manifestation of conditions characterized by insulin resistance. It can also be detectable earlier than fasting hyperglycaemia, the precise genetic, environmental and metabolic factors and sequence of events that lead to the underlying insulin resistance [7]. The excess in free fatty acids found in the insulin-resistant state is known to be toxic to hepatocytes. Putative mechanisms include high concentration of cell membrane disruption, toxin formation, mitochondrial dysfunction and inhibition and activation of key steps in the regulatory of metabolisms [8]. Other potential explanations for elevated transaminases in insulin-resistant states include peroxisomal beta-oxidation, recruited inflammatory cells and oxidant stress from reactive lipid peroxidation [9].

MATERIALS AND METHODS

Plant Collection- Fresh Lantana camara flowers were collected from [local nursery], identified and authenticated by a botanist- Dr. Sandeep K. Verma (Professor and Head of Dept. in Botany) SAGE University Indore (M.P.) [Voucher No. IOS /Bot/SLF-033]. [10]

Extraction -The flowers were shade-dried, powdered, and extracted with hydroalcoholic solvent (ethanol: water, 70:30) using Soxhlet apparatus. The extract was evaporated, dried, and stored at 4°C. Extraction method by Soxhletapparatus Around 500 gms dried flower of lantana camara lcoarsely powdered weighed and filled in Soxhlet apparatus for extraction. First the powdered drug was defatted with petroleum ether (Pet. Ether) (60°C-80°C); Defatted drug was then dried and again filled in soxhlet apparatus for successively extraction. Theextractionwascarriedoutforaperiodof72 extract obtained was dried in vacuum to remove excess solvent and were weighed for the determination of % yield11

Percentage yield estimation Dried extracts were calculated for percentage yield estimation using formula-

%yield=Practical yield /TheoreticalyieldX100

Lantana Camera linn Plant (flower) = 36.19 %

Acute Toxicity Study -The primary goal of the acute toxicity study was to determine the level of negative effects that the extract had on the experimental animal’s specific organs at an oral dose of 2000 mg/kg. The extract was safe at 2000 mg/kg, showing no deaths or behavioral changes, and is classified as low toxicity (LD?? > 2000 mg/kg) under OECD guidelines (2001). Additionally, a negligible variation in the weight of the vital organs of animals from the normal control group and the extract-treated group shows that extract did not produce any sensitivity, change, or acute organ damage.  Extracts preparation of flower LantanacamaraLinn.12

Preliminary phytochemical screening of the extract

Note: (-) Absence, (+) Presence and (++) present with high intensity of the colour.

Selection of animals

Wistar albino rats of either sex between 2 and 3 months of age weighing 150-200 g42 rats were used which were procured from the central animal house of College of Pharmacy, Swami Vivekanand College of Pharmacy, Indore (M.P.), India. All animals were housed in an animal room under normal condition of 25±1⁰C, 12-h light and dark cycle. The animals were allowed free to access commercial rat pallet diet and water. The bedding materials of the cages were changed every day. All the experimental procedures were carried out in accordance with the Committee for the Purpose of Control and Supervision of Experiments on Animals (IAEC) guidelines. IAEC Registration No. IAEC/SVCP/2025/Feb/01. The study designs were approved by the Institutional Animal Ethical Committee of College of Pharmacy,

Streptozotocin (STZ) induced diabetes in rats

After fasting 18 hours, the rats were injected intraperitoneal injection through tail vein with a single dose of 50 mg/kg Streptozocin, freshly dissolved in citrate buffer (pH 4.5). After injection, the rats had free access to food and water and were given 5% glucose solution to drink overnight to counter hypoglycemic shock²². Diabetes in rats was observed by moderate Polydipsia and marked Polyuria. The diabetes was confirmed by estimating the blood glucose level after 3 days by glucometer based one glucose oxidation method. Rats having blood glucose level more than 250mg/dl were selected for further study.13

Experimental Data

• Body Weight (g)- Body weight is are liable indirect marker of the metabolic state of diabetic rats. Improvement in body weight in treatment groups suggests restoration of insulin action and better glucose utilization.

Fasting Blood Glucose (mg/dL) In rat diabetes models, FBG > 200 mg/dL is considered diabetic. Reduction toward normal values indicates antidiabetic activity of the treatment.

• OGTT (mg/dL) OGTT in rats is an essential parameter to evaluate glucose tolerance and antidiabetic potential of drugs or plant extracts

• HbA1c (%) at Day 21- Diabetes in rats, HbA1c increases significantly due to chronic hyperglycemia. Reduction in HbA1c after treatment confirms sustained antihyperglycemic activity. HbA1c is often measured at the end of the study (e.g., 21 or 28 days

• Lipid Profile(mg/dL)- Lipid profile in diabetic rats is an essential parameter to assess antidiabetic and antihyperlipidemic activity of drugs plant extracts. Restoration toward normal values suggests cardioprotective potential.