Assessment of the Antidiabetic Properties of Essential Oil from Cannabis sativa

Received 28 Sep, 2023

Accepted 26 Dec, 2023

Published 31 Dec, 2023

This study used the hydrodistillation method to extract essential oil from Cannabis sativa (C.sativa) leaves, while its chemical composition was examined through gas chromatography-mass spectrometry. Experiments were conducted on alloxan-induced hyperglycemic rabbits (albino), with metformin as the reference medication for comparative analysis to evaluate its potential in managing diabetes. Chemical analysis showed that the principal components of C.sativa essential oil were caryophyllene (46.63%), d-limonene (7.123%) and cis-β-farnescene (9.115%), humulene (13.153%) and trans-α-bergamotene (6.592%). The minor components (˂5%) include α-pinene (3.720%), β-myrcene (1.910%), cineole (2.340%), valencene (1.614%), β-bisabolene (1.897%) and unidentified components (4.201-4.452%) were respectively. Intraperitoneal administration of C. sativa oil (0.7, 0.9 and 1.2µl/kg b.wt.) to hyperglycemia for 14 days resulted in a noteworthy decrease in both hepatic and fasting blood glucose levels. Meanwhile, there was a marked increase in the hepatic concentration of glycogen. The oil administration exhibited a reduced yet favourable anti-hyperglycemic potential compared to the reference antidiabetic drug. The study's findings indicate that the essential oil derived from  C. sativa leaves cultivated in Pakistan is characterized as a Beta Caryophyllene chemotype. Remarkably, the oil demonstrated significant efficacy in lowering glucose levels and showed promise in mitigating hyperglycemia-induced dyslipidemic complications in rabbits induced with alloxan.

INTRODUCTION

Diabetes mellitus is a persistent metabolic disorder marked by compromised insulin function or insufficient insulin production, which poses a substantial global health burden. As the prevalence of diabetes continues to rise, there is an escalating need for innovative therapeutic approaches to complement existing treatments(Campbell & Newgard, 2021; Sugandh et al., 2023; Shankar, 2023). IIn recent years, natural products derived from various plant sources have gathered attention for their potential antidiabetic properties. One such candidate is the essential oil extracted from C. sativa, commonly known as marijuana or hemp, has been traditionally employed for its medicinal properties. Beyond its well-documented psychotropic effects, this plant exhibits a rich chemical profile, with the essential oil representing a complex mixture of bioactive compounds (Chaachouay et al., 2023; Mittal et al., 2023; Rizzo et al., 2023). Preliminary studies suggest that certain constituents within the essential oil may possess antidiabetic attributes, making it a compelling subject for further investigation (Malabadi et al., 2023; Malabadi et al., 2023).

This research systematically evaluates the antidiabetic potential of the essential oil derived from C. sativa. By employing rigorous experimental methodologies and comprehensive analyses, we seek to elucidate the impact of essential oil on key markers of diabetes, including glucose metabolism and insulin sensitivity. Through this assessment, we aspire to contribute valuable insights into the therapeutic potential of C. sativa essential oil as a novel and natural intervention for managing diabetes. The findings from this study may deepen our understanding of the plant's medicinal properties and pave the way for the development of alternative and effective antidiabetic agents.

EXPERIMENTAL MATERIALS AND PROCEDURES

Instrument and chemicals
Major equipment used was a supercritical fluid extractor, spectrophotometer and centrifuge machine. All chemicals like Dimethyl sulfoxide (DMSO) employed in this study, meeting analytical-grade standards, were obtained from Merck.

Plant Material
In September-October 2022, fresh C.sativa plants were collected from District Chaniot Punjab, Pakistan. The plant specimens underwent further identification and authentication by a respected Taxonomist at the Department of Botany, University of Agriculture Faisalabad, Pakistan. The freshly collected leaf sample was promptly processed to extract essential oil (EO).

Isolation of Cannabis sativa essential oil
The extraction of essential oils from the leaves employed a batch process utilizing supercritical fluid at temperatures of 50°C and pressures of 90 bar. Each batch of supercritical fluid extraction utilized approximately 15kg of fresh C.sativa plant sample. Throughout the extraction process, continuous control of gas supply, pressure and temperature was maintained. The resulting oil yield was expressed as a percentage (% W/W). Post-extraction, The EOs were kept in vials made of black glass at temperatures between 0°C and 4°C until subjected to analysis.

Analysis of essential oil
A Perkin Elmer Clarus 600 gas chromatograph–mass spectrometer (GC–MS) system featuring an FID detector was used to analyze the essential oil. Helium gas was employed as the carrier, maintaining a consistent flow rate of ±1 ml/min. The mass transfer line and injector temperatures were set at 250°C and 300°C, respectively. The oven temperature was programmed to ramp up from 40°C (held for 2 minutes) to 270°C at a rate of 2°C/min, followed by an isothermal hold for 10 minutes, and ultimately increased to 300°C at a rate of 10°C/min. Diluted samples (1/100, v/v, in dichloromethane) of 1μl were injected in split mode with a split ratio of 120:1. The percentages of essential oil constituents were determined based on peak areas. Identification of the chemical constituents of C.sativa essential oil was achieved by analyzing GC retention time and calculating retention indices based on n-alkanes (C6–C24). Individual compounds were identified by comparing their mass spectra using NIST 2005 v.2.0 and Wiley Access Pak v.7 (2003) of GC–MS systems (Sahoo et al., 2022; Yan et al.,2023)

Preparation of Alloxan solution and standard drug
Alloxan monohydrate was dissolved in regular saline to form the alloxan solution. Glimepiride 1mg and Gum tragacanth were obtained from Shifa Pharmacy, Susan Road, Faisalabad. 2% suspension of Gum traganth was prepared by dissolving it in 100 ml water. Glimepiride was dissolved into this 2% gum suspension and made up to 10 ml volume.

Administration of Essential oil samples and standard drug
Essential oil doses of C. sativa
The extracted essential oil from C. sativa, obtained through supercritical fluid extraction at 50°C and 90 Bar pressure, was utilized. The quantity/volume of essential oil assigned to each albino rabbit was determined based on weight. The calculated volumes (0.7, 0.9, and 1.2 μl/kg) were meticulously triturated in 2 ml of a 2% gum tragacanth suspension, and the final volume was adjusted to approximately 5 ml by adding gum tragacanth suspension. This resulting suspension was then orally administered to each albino rabbit using a gastric tube connected to a 10 ml graduated syringe. The gastric tube was carefully introduced into the stomach via the esophagus, and the plunger was slowly pressed to administer the prescribed dose. Glimepiride was also administered by suspending it in 5 ml of a 2% gum tragacanth solution.

Induction of diabetes in Swiss albino rabbits
All groups, except group I, were induced into a diabetic state by intravenously injecting 150 mg/kg body weight of alloxan. Following the alloxan injection, all surviving rabbits' blood glucose (BG) levels were assessed using a BG testing kit. Adult albino rabbits exhibiting BG levels in the range of approximately 250-300 mg/dl were identified as diabetic and subsequently included in subsequent experimental studies (Baghel et al.,2023)

Grouping of animals
Eighteen albino rabbits in good health were selected and distributed randomly into four groups, as outlined in Table 1. Group IV was further divided into three subgroups (n=3) to facilitate the administration of different doses of C. sativa essential oil. Each rabbit had an average body weight within the range of 1.5-2kg. The rabbits underwent a one-week acclimatization period before the commencement of the experiment. Throughout the study, the animals were provided with regular seasonal fodder, and water was made available to adult albino rabbits at all times (Baghel et al.,2023).

Table 1:

Diet and administration timetable for adult albino rabbits throughout the 0-14 day experimental period

Gp #	Treatments	No of animals	Subgroups	Feeding and Drug Administration schedule
I	Untreated control of the routine feed		3	Routine normal feed 0 to 14 days
II	Untreated control on alloxan at 150 mg/kg body weight		3	Routine feed+ alloxan intravenously
III	Treated control on the synthetic antidiabetic drug: Glimepiride at 800μg/kg body weight orally		3	Routine feed + alloxan+ Glimepiride in 2% gum tragacanth as vehicle 4 to 14 days
IV	C. sativa E. oil SCFE at 50 C, 90 bar	I	3	Routine feed +alloxan++0.7μl/kg E. oil, 4to 14 days
		II	3	Routine feed +alloxan++0.9μl/kg E. oil, 4 to 15 days
		III	3	Routine feed +alloxan++1.2μl/kg E. oil, 4 to 15 days

Hypoglycemic Investigations
Blood Sample Collection
Blood samples were obtained from the jugular vein of each animal on days 0, 5, 10, and 15. In addition to the designated sampling days, samples were aseptically gathered at 0, 2, 4, 8, 16, and 24 hours on each scheduled sampling day. After clotting, serum was isolated via centrifugation and stored at 4ºC in a refrigerator.

Blood Glucose Measurement
The glucose level in the blood samples was assessed using the Kit method (glucose GOD-PAP, UK). The application of the glucose oxidase method ensured precise and reliable results. Currently, the glucose kit method is the most straightforward and extensively employed technique in this context.

Statistical Analysis
The results underwent evaluation through a Two-Way Analysis of Variance. The statistical distinctions between groups were examined using Duncan's Multiple Range Test at a 5% level of significance (Dash et al., 2023)

RESULTS AND DISCUSSION

Essential Oil Yield and Composition
Table 2 presents the yield of C. sativa essential oil obtained through supercritical fluid extraction. An optimal yield of 0.031 ± 0.02% was achieved at temperatures and pressures of 50°C and 90 Bar, respectively. The highest quality oil can be extracted at low temperatures (Yousefi et al., 2019). Current findings regarding the yield and explanation of supercritical fluid extraction (SCFE) closely align with the outcomes reported by (Rajput et al., 2023) for the extraction of essential oil from clove buds.

Table 2:

Extraction yield (%) of C. sativa essential oil by SCFE

Sr No.	Temperature and pressure	Weight (Kg)	Distilled water Vol. (litr)	Time of Extraction (hr)	Oil (g)	Yield %
1	50 ºC, 90 Bar	12	Nil	3hr	3.8	0.031±0.02a
2		12	Nil	3hr	3.76	0.029±0.01b
3		12	Nil	3hr	3.74	0.025±0.03c
Values are presented as the mean ± standard deviation of three samples. Superscript letters within the same column indicate significant differences at P ≤ 0.05 among the extractions of C. sativa EO

GC MS analysis of SCFE C. sativa essential oil
The chemical compositional data of the EOs from C. sativa essential oil obtained by SCFE are reported in Table (3) and showed in Fig.(1). Fifty compounds were identified in the EOs of C. sativa extracted at 50ºC, 90 bar extraction temperature and pressure respectively. At an extraction temperature of 50ºC and 80 Bar, the primary constituents of C. sativa essential oil included caryophyllene (46.63%), humulene (13.153%), d-limonene (7.123%), cis-β-farenscene (9.115%), and trans-α-bergamotene (6.592%). Additionally, the essential oils contained various minor components, with α-pinene (3.720%), β-myrcene (1.910%), cineole (2.340%), valencene (1.614%), β-bisabolene (1.897%), and unidentified components (4.201-4.452%) among them. Other components included many unidentified components (1.802–0.01%) and many trace components (˂1%), such as p-cymol, allo-aromadendrene, gamma-murolene (δ-murolene), α-cubebene etc. The overall identified constituents were categorized as monoterpene hydrocarbons (12 components) at 24.91%, sesquiterpene hydrocarbons (14 components) at 31.50%, oxy-monoterpenes (3 components) at 6.32%, oxy sesquiterpenes (1 component) at 2.27%, others (2 components) at 4.44%, and unidentified (UI) compounds (12 components) at 30.45% in the C. sativa essential oil extracted at 50°C, 90 bar.

The hydrocarbons identified in C. sativa encompass n-alkanes spanning from C9 to C39, including 2-methyl, 3-methyl, and certain dimethyl alkanes. In essential oils obtained through extraction and steam distillation, the primary alkane detected was the n-C29 alkane nonacosane, constituting 56.8% and 11.7%, respectively. Other notable alkanes included hentriacontane, heptacosane, pentacosane, 2, 6-dimethyltetradecane and hexacosane. The significant variation in essential oil compositions could be attributed to the extraction method employed and the extraction temperature (Pieracci et al., 2021).

Fig. 1: GC MS chromatogram of C. sativa SCFE EO at 50°C, 90 Bar

A diverse array of volatile compounds, spanning various chemical classes such as alcohols, ketones, hydrocarbons, and esters, has been identified in C. sativa samples. The volatile fraction's chemical composition in Cannabis-derived products has been thoroughly documented, primarily clarified through the analysis of their essential oils (Eržen et al., 2021; Rock et al., 2021). Aromatic compounds were extracted from C. sativa using supercritical CO₂ extraction coupled with online fractionation. The predominant constituents in the essential oil included terpinolene (6.33%), caryophyllene (31.24%), myrcene (11.83%), α-pinene (10.08%), α-humulene (9.75%), caryophyllene oxide (6.17%) and β-pinene (3.65%). The composition of the essential oil exhibited notable quantitative variations when compared to essential oils extracted from different fiber hemp inflorescences, as previously reported by (Mazzara et al., 2022). However, the presence of these primary constituents was verified. The essential oil demonstrated a higher percentage of sesquiterpenes (54.23%) and associated oxygenated compounds (12.64%) compared to hydrocarbon monoterpenes (35.31%) and oxygenated monoterpenes (1.24%).

Table 3:

Chemical composition of C. sativa essential oil obtained by SCFE at different temperature and pressure conditions

Peak #	Compound	RT	RI	50°C, 90 Bar
1	Α-Thujene	5.085	925.23	0.068±0.05b
2	Α-pinene	5.265	939.53	3.720±0.05a
3	Camphene	5.64	950.57	0.080±0.07b
4	Β-phellandrene	6.26	966.23	0.043±0.02a
5	Β-pinene	6.37	972.15	1.081±0.06a
6	Β-myrcene	6.71	915.62	1.910±0.01b
7	(+)-4-carene	7.516	978.11	0.178±0.12b
8	P-Cymol	7.762	1020.56	0.415±0.05a
9	D-limonene	7.906	1037.28	7.123±0.05a
10	Cineole	7.996	1040.34	2.340±0.06a
11	Trans-β-ocimene	8.172	1044.09	0.186±0.11a
12	Β-ocimene	8.521	1054.56	0.714±0.12a
13	Gamma-terpinene	8.906	1059.82	0.611±0.04b
14	(+)-4-carene	9.962	1062.11	0.118±0.07b
15	Nonanal	10.51	1063.23	0.023±0.02
16	Terpinen-4-ol	13.38	1178.21	0.047±0.06a
17	Ylangene	21.28	1181.21	0.064±0.09b
18	Copaene	21.47	1185.63	0.130±0.02c
19	U.i	22.72	1193.22	0.043±0.10c
20	U.i	23.08	1195.61	0.643±0.06a
21	U.i	23.09	1197.54	0.927±0.06b
22	Caryophyllene	23.27	1431.53	46.63±0.03a
23	U.i	23.56	1440.32	0.060±0.12c
24	(E)-β-famesene	23.72	1455.34	0.113±0.01
25	Trans-α bergamotene	23.89	1459.33	6.592±0.08a
26	U.i	24.17	1460.22	0.046±0.02a
27	U.i	24.19	1462.91	-
28	Aromandendrene	24.41	1463.22	0.189±0.04b
29	Humulene	24.58	1464.11	13.153±0.09a
30	Cis-β-farnesene	24.74	1465.34	9.115±0.05a
31	Alloaromadendrene	24.85	1467.53	0.373±0.08a
32	U.i	24.99	1473.44	0.060±0.01b
33	Gamma-muurolene	25.49	1487.73	0.396±0.04c
34	Β-curcumene	25.6	1491.32	0.188±0.05a
35	U.i	25.61	1493.56	-
37	U.i	25.86	1503.44	-
38	Valencene	26.22	1510.55	1.614±0.07a
40	U.i	26.43	1514.66	-
41	Α-Bulnesene	26.63	1515.69	0.149±0.03c
42	Β-bisabolene	26.74	1520.32	1.897±0.04a
43	U.i	26.94	1523.31	1.171±0.03b
44	U.i	27.32	1526.33	0.018±0.01c
45	U.i	27.33	1527.85	-
46	Naphthalene,1,2,3,4,4a,7-hexahydro-1,6-dimethyl-4-(1-methylethyl)-	27.63	1533.99	1.902±0.01a
47	1hcycloprop[e]azulene,1a,2,3,4,4a,5,6,7boctahydro1,1,4,7tetramethyl-, [1a (1a.α.,4.α.	27.85	1535.63	0.057±0.02c
48	U.i	28.05	1539.07	0.120±0.04c
49	Caryophyllene oxide	28.37	1543.21	0.892±0.12b
50	U.i	28.62	1544.54	0.186±0.10a
51	U.i	29.54	1546.22	0.025±0.01b
52	U.i	30.13	1547.13	0.029±0.04
53	U.i	30.51	1548.9	4.352±0.04a
54	U.i	31.24	1550.07	0.153±0.10c
55	U.i	33.55	1553.34	0.990±0.05a
56	U.i	31.24	1554.66	0.240±0.01b
57	U.i	33.54	1557.72	0.667±0.03a

	1.	Values represent the mean ± standard deviation of three samples of each C. sativa EO, individually analyzed in triplicate. Different superscript letters in the same row indicate significant differences (p<0.05)
	2.	Compounds are presented in the order of elution from the ULTRA Dimethyl polysiloxane-packed capillary column
	3.	Retention indices are relative to n-alkanes (C6–C24) on the MS Clarus 600 column
	4.	RT signifies identification based on retention time; RI signifies identification based on retention index; MS signifies identification based on the comparing mass spectra. t = traces<0.05

Determination of Antidiabetic activity
Diabetes mellitus is a metabolic disorder marked by hyperglycemia, glucosuria, and negative nitrogen balance. It primarily results from insufficient insulin secretion in the pancreatic beta cells and desensitization of insulin receptors. The prevalence of this condition has escalated to epidemic levels in the current century (Hichri et al., 2019). Currently, various medications like biguanides and sulfonylureas exist to alleviate hyperglycemia in diabetes mellitus. However, these drugs come with associated side effects. Hence, it is imperative to explore new compound classes to address these issues (Derici et al., 2021).C. sativa has been employed for various medicinal purposes, including the treatment of conditions like diabetes and as an early remedy for snakebites. Given its historical use in indigenous diabetes treatments, this study explores the potential of C. sativa essential oil and its isolated component fractions to improve insulin sensitivity (Kim et al., 2023; Sabir et al., 2020).

Hypoglycemic Investigation
The blood glucose levels were assessed in both standard and experimental rabbits, revealing a promising antidiabetic effect of C.sativa essential oil, as presented in Tables Tables (4 & 5) and Fig. (2-4). The outcomes in Table (4) demonstrate a significant reduction in fasting blood glucose levels (p < 0.05) over the 24-hour study period after administering a dose of 0.7 ml/kg glucose to the rabbits. In the case of chronic administration of alloxan, highly significant variations (p < 0.001) were observed between the investigational and diabetic control rabbits, leading to a substantial reduction in fasting blood glucose levels. Comparative analysis with the results of the standard antidiabetic drug exhibited significant variations (p < 0.05) and (p < 0.001). The doses of essential oil and isolated fractions 0.7ml/kg, 0.9ml/kg, and 1.2 ml/kg significantly lowered blood glucose levels in all days from 0 to 14 days and showed maximum reduction with 1.2 ml/kg, was found at 14 days. 1.2ml/kg dose showed maximum reduction of glucose 243.4 mg/100ml followed by 0.9ml/kg dose, 256.5mg/100ml> 0.7ml/kg, 258.75 mg/100ml respectively as shown in Table (4).

However, there is no previous report or data available about the antidiabetic potential of C. sativa essential oil (Akhtar et al., 2020). studied the in vivo impact of C. sativa extract on blood coagulation, fat metabolism, and glucose metabolism was investigated in both standard and streptozocin-induced diabetic rats, yielding promising results. In the liver tissue of both diabetic and normal rats, there was a general decrease in glycogen content, with C. sativa extract-treated rats exhibiting a significantly lower amount (P<0.001) compared to the untreated control rats.

In this study, alloxan was selected to induce a diabetic state in mice. Alloxan is a specific toxin known for its capacity to destroy pancreatic β-cells, leading to a primary deficiency of insulin without impacting other types of islets (Caixeta et al., 2020: Aju et al., 2019). Put differently, alloxan induces hyperglycemia by significantly diminishing insulin release through the destruction of the β-cells in the islets of Langerhans (Longkumer et al., 2021). Given the increasing inclination toward incorporating natural remedies alongside conventional treatments, plants traditionally employed may offer a valuable reservoir of potential new hypoglycemic compounds (Madariaga et al., 2023). Many authors have previously documented the potential antidiabetic properties of essential oils derived from medicinal plants (Nazir et al., 2021; Usman et al., 2020). Several plants have been identified to exhibit hypoglycemic effects. The proposed mechanisms underlying these effects involve the augmentation of insulin secretion from the β-cells of the islets of Langerhans or the liberation of bound insulin. In simpler terms, the hypoglycemic effects of plant essential oils may be ascribed to their insulin-mimicking actions (Top of FormBottom of Form Rocamora et al., 2020). t could be inferred that C.sativa similarly exerts its hypoglycemic activity. Therefore, the potential mechanism of action for the essential oil and isolated fractions could be linked to evocative impact akin to hypoglycemic sulfonylureas, which involves the facilitation of channels responsible for insulin secretion, membrane depolarization, and the initiation of Ca²⁺ influx. This signifies a pivotal initial stage in insulin secretion (Mukai et al., 2022; Campbell et al., 2021). The essential oil might additionally function by enhancing glucose utilization in peripheral tissues, particularly considering that alloxan treatment results in the permanent destruction of β-cells (Wariyapperuma et al., 2020: Patle et al., 2021).

Table 4:

Mean ±SEM levels of blood glucose expressed in mg/100ml at various time intervals at 0 day

Treatments	Doses	0hr	2hr	4hr	8hr	16hr	24hr	Total
Control		93.65	95.54	96.2	90.63	93.59	91.39	93.5
Alloxan at 150 mg/kg b.w		254.13	257.17	256.63	257.71	259	256.55	256.87c
Glimepiride at 800μg/kg b .w		235.7	238.6	241.42	244.59	248.68	251.09	243.3 b
E. oil (SCFE, 50°C)	0.7 ml/Kg	242.09	247.94	250.08	255.17	259.35	263.75	253.06
E.oil (SCFE, 50°C)	0.9 ml/Kg	237.84	245.69	247.83	252.92	257.1	261.5	250.48c
E.oil (SCFE, 50°C)	1.2ml/Kg	234.34	235.84	237.19	239.4	242.9	248.4	239.68a
Means sharing similar letters in a row or in a column is statistically non-significant (P>0.05). Small letters represent comparison among interaction means and capital letters are used for the overall mean

Table 5:

Mean ±SEM levels of blood glucose expressed in mg/100ml from day 0 to 14

Doses	Treated groups	0 day (mg/100mL)	5 days (mg/100mL)	10 days (mg/100mL)	14 days (mg/100mL)
	Control	93.5	93.39	91.39	91.39
	Alloxan at 150 mg/kg b.w	256.87	296.87	282.87	276.87
	Glimepiride at 800μg/kg b .w	243.35	269.09	256.09	251.09
0.7 ml/Kg	E.oil (SCFE, 50°C)	253.06**	272.75***	268.75***	258.75**
0.9 ml/Kg	E.oil (SCFE, 50°C)	250.48**	270.5***	266.5***	256.5**
1.2ml/Kg	E.oil (SCFE, 50°C)	239.68***	257.4**	253.4**	243.4***
Values are given as mean ± SEM, (n = 3); ** p<0.05 *** p<0.001, Duncan test as compared to diabetic control (Group-2)

Fig. 2: Blood glucose level of treated groups at 5th day

Fig. 3: Blood glucose level of treated groups at 10th day

Fig. 4: Blood glucose level of treated groups at 5th day

 

CONCLUSION

In conclusion, the investigation into the antidiabetic properties of the essential oil derived from C. sativa leaves has revealed promising outcomes. Through meticulous chemical analysis, the composition of the essential oil was elucidated, highlighting significant constituents such as caryophyllene, d-limonene, cis-β-farnescene, humulene, and trans-α-bergamotene, along with minor components including α-pinene, β-myrcene, cineole, valencene, β-bisabolene, and unidentified compounds. The experiment, conducted on alloxan-induced hyperglycemic rabbits over 14 days, demonstrated a significant decrease in both hepatic glucose levels and fasting blood glucose following the intraperitoneal administration of C. sativa oil. Additionally, a notable increase in hepatic glycogen concentration was observed. While the anti-hyperglycemic potential of the oil was slightly reduced compared to the reference antidiabetic drug metformin, it nevertheless exhibited a favourable impact. The study classifies the essential oil as a Beta Caryophyllene chemotype, sourced explicitly from Cannabis sativa leaves cultivated in Pakistan. Markedly, the oil demonstrated substantial efficacy in lowering glucose levels and showed promise in addressing hyperglycemia-induced dyslipidemia complications. These findings underscore the potential of C. sativa essential oil as a candidate for further exploration in the quest for effective strategies in the management of diabetes. Continued research is warranted to elucidate the underlying mechanisms and to ascertain their viability for clinical applications.

ACKNOWLEDGEMENT

We want to thank all the authors who contributed to this paper. Their expertise, insights, and feedback were invaluable for shaping the research question, methodology, analysis, and discussion. We also appreciate their patience and cooperation throughout the writing and revision process. Without their support and collaboration, this paper would not have been possible.

CONFLICT OF INTEREST

No conflict of interest exists among the authors

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