Comparative Evaluation of Apoptosis Induction Using Needles, Bark, and Pollen Extracts and Essential Oils of Pinus eldarica in Lung Cancer Cells

Lung cancer is one of the leading causes of cancer-related mortality worldwide. Although effective clinical drugs for treating advanced stages are available, interest in alternative herbal medicines has gained momentum. Herbal extracts are potent antioxidants that reportedly inhibit the growth of various cancer cell lines. In the present study, we investigated the effects of essential oils and hexane, methanolic, and aqueous extracts, obtained from various parts (bark, needles, and pollen) of Pinus eldarica against human lung cancer (A549) cells. First, the DPPH radical scavenging activities of P. eldarica extracts and essential oils were examined, which revealed that methanolic extracts presented higher antioxidant activity than the other extracts and essential oils. Next, A549 cells were exposed to various concentrations of the extracts and essential oils for 48 h. P. eldarica extracts/essential oil-treated lung cancer cells demonstrated a significant decrease in cell proliferation, along with an induction of apoptotic cell death, particularly, the pollen hexane extract, bark essential oil, and methanolic needle extract showed superior results, with IC50 values of 31.7, 17.9, and 0.3 μg/mL, respectively. In the cell cycle analysis, treatment of A549 cells with the methanolic needle and pollen hexane extracts led to apoptosis and accumulation of cells in the sub-G1 phase. Further, exposure to the bark essential oil and methanolic needle extract decreased the cell population in the G2/M phase. Notably, treatment with the pollen hexane extract, bark essential oil, and methanolic needle extract resulted in caspase-3 activation, poly (ADP-ribose) polymerase cleavage, Bcl-2 downregulation, and Bax and p53 regulation in A549 cells. Furthermore, these extracts and essential oils decreased the migration, and colony formation of A549 cells. These findings provide experimental evidence for a new therapeutic effect of P. eldarica against human lung cancer and suggest P. eldarica as a potential chemopreventive natural resource for developing novel cancer therapeutics.

Antioxidant Effects of Pinus eldarica Extracts Against Cisplatin-induced Cytotoxicity in Human Hepatoma (HepG2) Cell Line

Background: Cisplatin has cytotoxic effects through free radical generation. Reports indicate that various members of Pinaceae family may have antioxidant properties. In this study we investigated the cytoprotective effects of needle volatile oil and bark extract of Pinus eldarica in Hepatoma G2 (HepG2) cell line. Methods: Using the maceration technique, we obtained the ethanolic extract of Pinus eldarica’s bark (BHAEPE). Folin-ciocalteu reagent was used to determine the total phenolic content of BHAEPE. The Needle’s Volatile Oil of Pinus eldarica (NVOPE) was obtained by Clevenger hydrodistillation method. The main components of NVOPE were identified by Gas Chromatography and Mass Spectrophotometry (GC-MS). We used DPPH assay to investigate the antioxidant activity of both NVOPE and BHAEPE. Also, MTT assay was performed to test the protective effects of both BHAEPE and NVOPE against cisplatin. Results: Folin-ciocalteu test demonstrated that each gram of BHAEPE was equivalent to 371±6 mg of gallic acid. Also, GC-MS identified germacrene D as the main component of NVOPE. BHAEPE had more antioxidant capacity compared to NVOPE. When incubated solely with cells, neither BHAEPE nor NVOPE represented cytotoxicity on. Furthermore, BHAEPE demonstrated mitogenic effects on higher doses (50, 75 and 100 μg/ml). However, both NVOPE and lower doses of BHAEPE (10, 20 μg/ml) could not protect the cells against cisplatin’s cytotoxicity, but higher doses of BHAEPE provided cytoprotective effects. Conclusion: The antioxidant properties of both NVOPE and lower doses of BHAEPE did not adequately protect HepG2 cells against the cisplatin’s cytotoxicity. However, BHAEPE at high concentrations demonstrated cytoprotective effects.

Evacuation the Effect of Single- and Multi-dose Administration of Ethanolic Extract of Pinus eldarica Pollen Against Acetaminophen-induced Rat Liver Injury

Introduction: Acetaminophen (APAP)-induced liver injury is one of the main causes of acute liver failure in the world. Pinus eldarica is specially distributed in the north of Iran and has been used for decades to treat wounds, pain, infection, fever, bronchitis and inflammation in many countries. This study was undertaken to evaluate the role of pine pollen extract (PE) on APAP-induced hepatotoxicity. Methods: This study was conducted in two separate parts: single- and multi- dose administration of PE. In a multiple dosing regimen, different doses of extract (10, 20, 30, 40 and 50 mg/kg) were orally administrated for 2 weeks and then on the 15th day, the animals received a single dose of APAP (600 mg/kg). In the second part, the highest dose of PE (50 mg/kg) was orally administrated half an hour after receiving the APAP at dose of 600 mg/kg. The negative and positive control groups were treated with normal saline and N-acetylcysteine (NAC), respectively. At the end of procedure, the biochemical parameters including alanine transaminase (ALT), aspartate transaminase (AST) and alkaline phosphatase (ALP) as well as pathological findings were evaluated. Results: A single and multiple oral dose of APAP increased the serum level of ALT, AST and ALP that were significantly attenuated by PE administration in both model. Indeed, exposure to APAP caused extensive necrosis and lymphocytic inflammation that were completely prevented by single- and multi-dose administration of PE, specially at a high dose. Conclusion: Finally, the present results indicate that PE could reverse the reduction of aminotransferases and improve histological changes of APAP-induced liver toxicity. Further research is needed to confirm this finding and reveal the exact mechanisms.

PINUS ELDARICA MEDW. AS INDICATOR OF VEHICULAR POLLUTION

This paper presents a study of developmental stability of Pinus eldarica Medw. in conditions of airborne traffic pollution. The study has been performed over the period of 3 years (from 2017 to 2019) in the Absheron Peninsula, Azerbaijan. The purpose of the study was to assess the morphophysiological changes in the species Pinus eldarica Medw. Exposed to pollution from motor vehicles and thus identify the bioindicator properties of pine trees. The study used morphological, physiological and statistical methods. The indicating data on fluctuating asymmetry and necrosis in Pinus eldarica Medw. was obtained in areas with varying degree of atmospheric pollution. Based on these data, ecological zoning was carried out using GIS technologies, and an environmental map of the studied territories was compiled. Species Pinus eldaric a Medw. can be used as a phytometer for the purpose of environmental monitoring.

Chemical Composition and Antimicrobial Activity of Essential Oils from the Aerial Parts of Pinus eldarica Grown in Northwestern Iran

Pinus eldarica (Pinaceae), an evergreen plant, is distributed across the warm and dry climates of western Asia, including Asia Minor, the Middle East, and land surrounding the Caspian Sea. Essential oils (EOs) from different aerial parts of this tree have been used in traditional medicine. We aimed to investigate the chemical profile and antimicrobial activity of the EO from P. eldarica grown in northwestern Iran. EO from the needles, bark, and pollen were extracted with boiling water using a Clevenger apparatus at yield of 0.7–1.2 cm3/100 g of dry plant material. The main chemical components of the EO from the needles were D-germacrene (18.17%), caryophyllene (15.42%), γ-terpinene (12.96%), and β-pinene (10.62%); those from the bark were limonene (16.99%), caryophyllene oxide (13.22%), and drimenol (13.2%); and those from the pollen were α-pinene (25.64%) and limonene (19.94%). In total, 83 constituents were characterized in the EOs, using gas chromatography mass spectrometry analysis; mainly, sesquiterpene hydrocarbons in needle EO and monoterpene hydrocarbons in pollen and bark EOs. β-Pinene, β-myrcene, limonene, and caryophyllene were identified in the EOs from all three plant parts. The antibacterial and antifungal properties of the EOs were examined: pollen EO exhibited antibacterial activity against Escherichia coli; bark EO inhibited the growth of Candida albicans and Staphylococcus aureus; and the needle EO inhibited the growth of S. aureus. Thus, the EOs from aerial parts of P. eldarica can benefit the EO industry and antibiotic development.