Exploring the Chemical Constituents, Antioxidant, Xanthine Oxidase and COX Inhibitory Activity of Commiphora gileadensis Commonly Grown Wild in Saudi Arabia.

The use of the synthetic drugs has increased in the last few decades; however, these drugs exhibit various side effects. Scientists are therefore seeking alternatives from natural sources. Commiphora gileadensis has long been used to treat various disorders. It is commonly known as bisham or balm of Makkah. This plant contains various phytochemicals, including polyphenols and flavonoids, with biological potential. We found that steam-distilled essential oil of C. gileadensis exhibited higher antioxidant activity (IC50, 22.2 µg/mL) than ascorbic acid (IC50, 1.25 µg/mL). The major constituents (>2%) in the essential oil were ß-myrcene, nonane, verticiol, ß-phellandrene, ß-cadinene, terpinen-4-ol, ß-eudesmol, α-pinene, cis-ß-copaene and verticillol, which might be responsible for the antioxidant and antimicrobial activity against Gram-positive bacteria. The extract of C. gileadensis exhibited inhibitory activity against cyclooxygenase (IC50, 450.1 µg/mL), xanthine oxidase (251.2 µg/mL) and protein denaturation (110.5 µg/mL) compared to standard treatments, making it a viable treatment from a natural plant source. LC-MS analysis revealed the presence of phenolic compounds such as caffeic acid phenyl ester, hesperetin, hesperidin, chrysin and transient amounts of catechin, gallic acid, rutin and caffeic acid. The chemical constituents of this plant can be explored further to investigate its wide variety of therapeutic potential.

Antibacterial Effects of Commiphora gileadensis Methanolic Extract on Wound Healing.

Commiphora gileadensis (CG) is a small tree distributed throughout the Middle East. It was traditionally used in perfumes in countries in this area. In Saudi Arabia, it was used to treat wounds burns and as an antidote to scorpion stings. This study aimed to evaluate the antimicrobial activity and cutaneous wound healing efficiency of the CG extracts using microbiological tests, rate of wound contraction and histopathological changes. CG plant were extracted using the methanol extraction technique; then, the methanolic extract was characterized using liquid chromatography coupled with mass spectrometry (LC−MS). Afterwards, a six-millimetre (mm) excision wound was induced in 60 male Balb/c mice. Mice were classified into two classes; each class consisted of three groups of 10 mice. In the non-infected wound class, the group I was assigned as control and received normal saline. Group II received gentamicin treatment, and group III treated with CG-methanolic extract. In the Staphylococcus aureus-infected class, group IV received normal saline, and groups V and VI were treated with gentamicin and CG-methanolic extract, respectively. The colonization of infected wounds was determined using colony-forming units (CFUs), and the percentage of wound contraction was measured in all groups. Finally, the histopathologic semi-quantitative determination of wound healing was evaluated by inflammatory cell infiltration, the presence of collagen fibres and granulation tissue, and the grade of re-epithelization. Composition analysis of the methanolic extract confirmed the presence of a high amount of ceramide (69%) and, to a lesser extent, hexosylceramide (18%) and phosphatidylethanolamine (7%) of the total amount. Additionally, there was a statistically significant difference between the percentage of wound contraction in the CG-treated and control groups in both Staphylococcus aureus-infected and non-infected wounds (p < 0.01). The colonization of the infected wounds was lower in the group treated with CG than in the control group (p < 0.01). In both non-infected and infected wounds, the CG-treated group showed significant statistical differences in inflammatory cell infiltration, collagen fibres, re-epithelization and granulation tissue formation compared with the control group (p < 0.01). The CG extract possesses antibacterial and anti-inflammatory properties that induce wound healing.

Forgotten Perfumery Plants - Part I: Balm of Judea.

Perfumes have always been products of great importance, mainly composed of natural, valuable and vegetal raw materials. Today, some of them have completely disappeared in perfumery, even though they are part of our cultural heritage and were commonly used in the past. Balm of Judea is one of the most noble, rare and fascinating ingredient long used in perfumery and medicine, that is missing today. After years of research, we collected a resin and an essential oil (steam distillation of fresh aerial parts) from Commiphora gileadensis (L.) C.Chr. native from Saudi Arabia and cultivated in Israel. The aims of this study were to i) identify the main reasons of the loss of the balm of Judea, ii) characterize the volatile composition of the resin and the essential oil and iii) evaluate their olfactory profile and assess their biological activity. Eighty-three compounds were identified in the resin, by a combination of GC-MS and GC/FID techniques, using direct injection and HS-SPME. α-Pinene (24.0 %), sabinene (43.8 %), ß-pinene (6.3 %) and cymene (3.6 %) were the main identified compounds, giving an intense, terpenic and lemony smell to the resin. Anti-inflammatory, wound-healing and whitening activities were highlighted. Sabinene (22.7 %), terpinen-4-ol (18.7 %), α-pinene (14.4 %) and cymene (13.6 %) were identified as the main components of the essential oil, giving a spicy, woody and lemony fragrance. Anti-inflammatory and whitening activities were emphasized.

Anticoagulative activity of Commiphora gileadensis, aspirin, and heparin on blood coagulation profiles in naïve mice.

Objective: Commiphora gileadensis is a small tree under the genus Commiphora. Previous studies showed medical applications, such as antibacterial and antihypertensive, for C. gileadensis. Methods: Sixty naïve mice were classified into six groups: control, C. gileadensis sap-treated group, C. gileadensis methanol extract-treated group, C. gileadensis acetone extract-treated group, heparin-treated group, and aspirin-treated group. Blood samples from each mouse in the six groups were collected in EDTA, sodium citrate, and heparin tubes. The body weight of each mouse was measured at the beginning and end of the experiment. Furthermore, complete blood count, kidney and renal function tests, coagulation profiles, prothrombin time (PT), activated partial thromboplastin time (aPTT), international normalized ratio (INR), D-dimer, and fibrinogen concentrations were estimated for each mouse. Results: The sodium, potassium, chloride, blood urea nitrogen, creatinine, alanine transaminase, and aspartate transaminase levels did not show statistical differences between all groups. Moreover, PT, aPTT, and INR were prolonged in the C. gileadensis sap, methanol, and acetone extracts-treated mice compared with those in the heparin and aspirin-treated groups (P < 0.01). D-dimer and fibrinogen concentrations did not show significant statistical differences between all groups. Conclusion: The current study concludes that the C. gileadensis sap, methanol, and acetone extracts prolonged PT, aPTT, and bleeding time in naïve mice more than heparin and aspirin. This means that the C. gileadensis extracts may have antithrombotic activity and may be used in the future to resolve intravascular thrombosis in patients having prosthetic valves.

UPLC-qTOF-MS phytochemical profile of Commiphora gileadensis leaf extract via integrated ultrasonic-microwave-assisted technique and synthesis of silver nanoparticles for enhanced antibacterial properties.

The utilization of metallic nanoparticles in bio-nanofabrication holds significant potential in the field of applied research. The current study applied and compared integrated ultrasonic-microwave-assisted extraction (US/MICE), ultrasonic extraction (USE), microwave-assisted extraction (MICE), and maceration (MAE) to extract total phenolic content (TPC). In addition, the study examined the antioxidant activity of Commiphora gileadensis (Cg) leaf. The results demonstrated that the TPC of US/MICE exhibited the maximum value at 59.34 ± 0.007 mg GAE/g DM. Furthermore, at a concentration of 10 µg/mL, TPC displayed a significant scavenging effect on DPPH (56.69 %), with an EC50 (6.48 µg/mL). Comprehensive metabolite profiling of the extract using UPLC-qTOF-MS/MS was performed to identify active agents. A total of 64 chromatographic peaks were found, out of which 60 were annotated. The most prevalent classes of metabolites found were polyphenols (including flavonoids and lignans), organic compounds and their derivatives, amides and amines, terpenes, and fatty acid derivatives. Transmission electron microscopy (TEM) revealed the aggregate size of the synthesized nanoparticles and the spherical shape of C. gileadensis-mediated silver nanoparticles (Cg-AgNPs). The nanoparticles had a particle size ranging from 7.7 to 42.9 nm. The Cg-AgNPs exhibited more inhibition zones against S. aureus and E. coli. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of Cg-extract, AgNPs, and Cg-AgNPs were also tested. This study demonstrated the feasibility of using combined ultrasonic-microwave-assisted extraction to separate and extract chemicals from C. gileadensis on a large scale. These compounds have potential use in the pharmaceutical industry. Combining antibacterial and biocompatible properties in materials is vital for designing new materials for biomedical applications. Additionally, the results showed that the biocompatibility of the Ag-NPs using C. gileadensis extracts demonstrated outstanding antibacterial properties.

Population Genetics, Genetic Structure, and Inbreeding of Commiphora gileadensis (L.) C. Chr Inferred from SSR Markers in Some Mountainous Sites of Makkah Province.

Commiphora gileadensis (L.) C. Chr is a perennial plant existing mainly in the southern and western mountains of the Arabian Peninsula. In the Makkah province, the remaining populations are threatened by many factors such as overcutting, overgrazing, and urban developments. These dangers are expected to be aggravated by the progression of aridification factors arising from climate change. To overcome the decline in remaining populations of this valuable species, a timely evaluation of the population's genetic variables and genetic structure is vital for the conservation of existing C. gileadensis populations. In this study, we used 61 SSR primers to achieve this objective. Only 50 loci showed polymorphisms, which led to further analysis of the population genetics for 600 genotypes that were collected from 50 populations of C. gileadensis found in 10 different sites in the Makkah region: Gebel Al Muliesaa, Wadi Albathna, Wadi Houra, Wadi Albaidaa, Wadi Elebiedia, Gebel Kniethl, Wadi Sayaa, Wadi Elbarasa, Wadi Alfawara, and Wadi Alkharar. The results showed an obvious decrease in genetic diversity variables in all studied populations. The range of PPL was between 8 and 40; additionally, the low HT value of 0.804 and the high value of inbreeding, Fis = 0.238, reflected a severe lack of heterozygotes. High levels of FST and GST and low gene flow indicate considerable segregation among the C. gileadensis populations, which creates a barrier to gene migration. Our data suggest the need for conservation planning for C. gileadensis in order to avoid the species' forthcoming extinction. Efforts should be largely oriented around managing water consumption, prohibiting overcutting and overgrazing, and establishing appropriate seed banks.

Antioxidant activity and total phenolic compounds of Commiphora gileadensis extracts obtained by ultrasonic-assisted extraction, with monitoring antiaging and cytotoxicity activities.

Commiphora gileadensis (C. gileadensis) has been identified and linked with various health benefits and pharmaceutical potential for its phytochemical activities and chemical constituents. This study aimed to evaluate ultrasonic-assisted extraction (USE) technique for total phenols content from C. gileadensis leaf compared to the hydrodistillation extraction (HDE). Our results showed that the USE operating conditions were identified as: MeOH·H2O solvent-to-fresh sample ratio of 80:20 (v/v); ultrasonic power/frequency at 150 W/20 kHz; and a temperature of 40 ± 1°C; subjected to acoustic waves intermittently for a calculated time (5 min) during the total programmed time of 12 min. The USE exhibited (118.71 ± 0.009 mg GAE/g DM) more amounts of all phenols than HDE (101.47 ± 0.005 mg GAE/g DM), and antioxidant (77.78 ± 0.73%, 75.27 ± 0.59% scavenging inhibition of DPPH), respectively. Anti-aging and Cytotoxicity activities were investigated. The results of biological evaluations showed that the crude extracts of C. gileadensis significantly extended the replicative lifespan of K6001 yeast. In addition, in vitro cytotoxicity against the HepG2 cell line showed significant anticancer activity, and approximately 100 µg/mL is required to decrease viability compared with that of the control. This study is proven for a larger scale to extract and isolate compounds of C. gileadensis for potential utilization in the pharmaceutical industry. In conclusion, advanced methods afford an extract with high activity in the biological properties of the extract.

Secondary Metabolites Profiling, Antimicrobial and Cytotoxic Properties of Commiphora gileadensis L. Leaves, Seeds, Callus, and Cell Suspension Extracts.

Commiphora gileadensis L. is an important endangered medicinal plant that belongs to the family Burseraceae. In this study, C. gileadensis callus culture was established successfully using mature leaves as explants cultured on Murashige and Skoog (MS) media supplemented with 24.50 µM of indole butyric acid (IBA) and 2.22 µM 6-Benzylaminopurine (BAP) (callus induction media). The obtained callus was maintained on MS medium supplemented with 16.11 µM naphthalene acetic acid (NAA) in combination with 6.66 µM BAP, which resulted in a substantial increase in callus fresh and dry weights. The cell suspension culture was established successfully using liquid callus induction media supplemented with 3.0 mg·L-1 proline. Thereafter, the chemical constituents of different C. gileadensis methanolic extracts (callus, cell suspension, leaves, and seeds) were profiled, and their cytotoxic and antimicrobial properties were investigated. The LC-MS GNPS analyses were applied for chemical profiling of the methanolic plant extracts, and several natural products were identified, including flavonols, flavanones, and flavonoids glycosides, with two unusual families that included puromycin, 10-hydroxycamptothecin, and justicidin B. The methanolic extracts have shown selective antimicrobial and cytotoxic properties against different microbes and cancer cell lines. For instance, leaf extract showed the highest zone of inhibition for Staphylococcus aureus, while cell suspension culture was effective against Staphylococcus epidermidis and Staphylococcus aureus. All extracts showed selective activity against A549 cell lines for the cytotoxicity assay, while the leaf extract had a broad cytotoxic effect against all tested cell lines. This study revealed that C. gileadensis callus and cell suspension cultures can be employed to increase the in vitro formation of biologically active compounds that may have cytotoxicity and antibacterial action against different cancer cell lines and bacterial species. Further studies are required to isolate and identify such constituents that corroborate the observed activities.

Genetic Characterization of Some Saudi Arabia's Accessions from Commiphora gileadensis Using Physio-Biochemical Parameters, Molecular Markers, DNA Barcoding Analysis and Relative Gene Expression.

Commiphora gileadensis L. is a medicinal plant, known as balsam, with pharmaceutical potential for its phytochemical activities and chemical constituents. Genetic diversity is a genetic tool used in medicinal plant evolution and conservation. Three accessions from C. gileadensis were collected from three localities in Saudi Arabia (Jeddah, Jizan and Riyadh). Genetic characterization was carried out using physio-biochemical parameters, molecular markers (inter-simple sequence repeat (ISSR) and start codon targeted (SCoT)), DNA barcoding (18 S rRNA and ITS rDNA regions), relative gene expressions (phenylalanine ammonia-lyase 1 (PAL1), defensin (PR-12)) and pathogenesis-related protein (AFPRT). The results of this study showed that C. gileadensis accession C3, collected from Riyadh, had the highest content from the physio-biochemical parameters perspective, with values of 92.54 mg/g and 77.13 mg/g for total phenolic content (TPC) and total flavonoid content (TFC), respectively. Furthermore, the highest content of antioxidant enzyme activity was present in accession C3 with values of 16.87, 60.87, 35.76 and 27.98 U mg-1 for superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) (mol/min/mg FW) and ascorbate peroxidase (APX) (U mg-1 protein), respectively. The highest total number of bands and number of unique bands were 138 and 59, respectively, for the SCoT marker. The SCoT marker was the most efficient for the genetic diversity of C. gileadensis by producing the highest polymorphism (75.63%). DNA barcoding using 18 S and ITS showed the nearby Commiphora genus and clustered C. gileadensis accessions from Jeddah and Jizan in one clade and the C. gileadensis accession from Ryiadh in a separate cluster. Moreover, relative gene expression of the PAL1, defensin (PR-12) and AFPRT (PR1) genes was upregulated in the C. gileadensis accession from Ryiadh. In conclusion, ecological and environmental conditions in each locality affect the genomic expression and genetic diversity, which can help the evolution of important medicinal plants and improve breeding and conservation systems.

In vitro and in vivo Antibacterial effect of Commiphora gileadensis Methanolic Extract against Methicillin-Resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa.

BACKGROUND AND OBJECTIVE: Commiphora gileadensis is a plant in the Burseraceae family that grows in the western area of Saudi Arabia. Traditionally, it is used in the treatment of some superficial infections. MATERIALS AND METHODS: The methanolic extract of Commiphora gileadensis isolated from its leaves and branches. The in vitro study was conducted to determine the effect of this extract on Methicillin-Resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa using an agar diffusion and Minimum inhibitory concentration (MIC) methods. The in vivo study was conducted through two different methods. The first method, 20 male Balb c-1 mice were used for the determination of Commiphora gileadensis methanolic extract toxicity (LD50). In the second method, 40 male mice were used and were put into four groups. The first and second groups were injected subcutaneously with 108 CFU of MRSA 1 mL-1, while the third and fourth groups were injected with 108 CFU of Pseudomonas aeruginosa 1 mL-1. The comparison between groups was done by using a t-test (p<0.05). RESULTS: The methanolic extract of Commiphora gileadensis had a greater sensitivity zone on MRSA and Pseudomonas aeruginosa, 7 and 3 mm respectively. The MIC of the extract was 1/8 and 1/2 for MRSA and Pseudomonas aeruginosa respectively. The in vivo study showed that the extract was non-toxic, it also showed that the extract decreased the mortality of mice induced by MRSA injection significantly (p<0.05) While insignificantly with Pseudomonas aeruginosa. CONCLUSION: The total Commiphora gileadensis methanolic extract had an antibacterial effect on MRSA and Pseudomonas aeruginosa. This extract was non-toxic for the mice.