Ivan Pavlović

Chloroform extract of underground parts of Ferula heuffelii: secondary metabolites and spasmolytic activity.

Plants from the genus Ferula L. (Apiaceae) were used for various purposes in traditional medicine of different nations throughout the history. Ferula heuffelii Griseb. ex Heuffel is a perennial species endemic for Balkan peninsula. Ten compounds which belong to classes of prenyl‐furocoumarin‐, prenyl‐dihydrofurochromone‐, prenyl‐benzoyl‐ and prenyl‐benzoylfuranone‐type sesquiterpenoids, as well as sesquiterpene coumarins and phenylpropanoids, were, for the first time, isolated from the CHCl3 extract of the underground parts of this plant and identified. Furthermore, extract and three isolated compounds, i.e., latifolone (1), dshamirone (4), and (2S*,3R*)‐2‐[(3E)‐4,8‐dimethylnona‐3,7‐dien‐1‐yl]‐2,3‐dihydro‐7‐hydroxy‐2,3‐dimethylfuro[3,2‐c]coumarin (6) were, for the first time, evaluated for their in vitro antispasmodic activities in three experimental models: spontaneous contraction, and ACh‐ and KCl‐induced contraction of an isolated rat ileum. The extract (0.1–1.3 mg/ml) and compound 6 (1–10 μg/ml) exhibited dose‐dependent effect in all three models. Compound 1 (1–6 μg/ml) affected spontaneous contractions and those induced by KCl, while compound 4 (8 μg/ml) displayed only moderate activity with ACh‐induced contractions. It can be concluded that tested compounds contribute to exhibited antispasmodic activity of crude extract. Additionally, extract (0.1–1.3 mg/ml) was tested for in vitro relaxant activity on an isolated rat trachea, and relaxed the KCl‐induced contractions in a dose‐dependent manner.

Antimicrobial and Cytotoxic Activity of Extracts of Ferula heuffelii Griseb. ex Heuff. and Its Metabolites

The antimicrobial and cytotoxic activities of isolates (CHCl3 and MeOH extracts and selected metabolites) obtained from the underground parts of the Balkan endemic plant Ferula heuffelii Griseb. ex Heuff. were assessed. The CHCl3 and MeOH extracts exhibited moderate antimicrobial activity, being more pronounced against Gram‐positive than Gram‐negative bacteria, especially against Staphylococcus aureus (MIC=12.5 μg/ml for both extracts) and Micrococcus luteus (MIC=50 and 12.5 μg/ml, resp.). Among the tested metabolites, (6E)‐1‐(2,4‐dihydroxyphenyl)‐3,7,11‐trimethyl‐3‐vinyldodeca‐6,10‐dien‐1‐one (2) and (2S*,3R*)‐2‐[(3E)‐4,8‐dimethylnona‐3,7‐dien‐1‐yl]‐2,3‐dihydro‐7‐hydroxy‐2,3‐dimethylfuro[3,2‐c]coumarin (4) demonstrated the best antimicrobial activity. Compounds 2 and 4 both strongly inhibited the growth of M. luteus (MIC=11.2 and 5.2 μM, resp.) and Staphylococcus epidermidis (MIC=22.5 and 10.5 μM, resp.) and compound 2 additionally also the growth of Bacillus subtilis (MIC=11.2 μM). The cytotoxic activity of the isolates was tested against three human cancer cell lines, viz., cervical adenocarcinoma (HeLa), chronic myelogenous leukemia (K562), and breast cancer (MCF‐7) cells. The CHCl3 extract exhibited strong cytotoxic activity against all cell lines (IC50<11.0 μg/ml). All compounds strongly inhibited the growth of the K562 and HeLa cell lines. Compound 4 exhibited also a strong activity against the MCF‐7 cell line, comparable to that of cisplatin (IC50=22.32±1.32 vs. 18.67±0.75μM).

SUPEROXIDE DISMUTASE AND LIPID PEROXIDATION IN CHILDREN AFFECTED BY CELIAC DISEASE

Celiac disease (CD) is an autoimmune disorder provoked by the wheat gluten and related proteins from other grains. The only treatment for the patients is a lifelong gluten free diet (GFD). Oxidative stress has been implicated in the pathogenesis of CD. The aim of this study was to examine the modulation of the biochemical response to oxidative stress in children affected by CD. The study involved peripheral blood samples and small intestinal biopsies from 69 children diagnosed with CD. According to the histological findings, patients were divided into following groups: Marsh 0: normal mucosa with no signs of inflammation (n=31); Marsh 1: mucosa was characterized by intraepithelial lymphocytosis (n=5); Marsh 2: intraepithelial lymphocytosis was accompanied by crypt hyperplasia (n=4); Marsh 3a: mucosa showed partial villous atrophy (n=20); Marsh 3b: subtotal villous atrophy was present (n=9). For the statistical purposes groups Marsh 1 and Marsh 2 were treated as one (Marsh 1+2, n=9). The activities and protein levels of copper, zinc superoxide dismutase (CuZnSOD) and manganese SOD (MnSOD), as well as the concentrations of lipid hydroperoxides (LOOH) were determined in intestinal biopsies, while in the peripheral blood, MnSOD activity was not measured, due to the methodological obstacles. CuZnSOD activity in the blood varied significantly between the analyzed groups. Marsh 3a and Marsh 3b had increased CuZnSOD activity comparing to the Marsh 0 (P < 0.05). LOOH concentration also varied significantly. LOOH level was higher in the blood of Marsh 3a (P < 0.001) and Marsh 3b (P < 0.05), than in Marsh 0. In the biopsy samples, MnSOD activity and LOOH concentration showed significant differences between the groups, while no significant difference was found for CuZnSOD activity. In comparison to Marsh 0, MnSOD activity was significantly elevated in Marsh 3a (P < 0.01). Significant increase in LOOH concentration was found in Marsh 3a (P < 0.001) and Marsh 3b (P < 0.01), comparing to Marsh 0. In addition, Marsh 3a group had higher LOOH concentration than Marsh 1+2. Relative MnSOD and CuZnSOD protein level in the peripheral blood and intestinal mucosa did not vary significantly between the analyzed groups. Positive correlations were found between the severity of mucosal lesion and CuZnSOD activity (P < 0.001), as well as LOOH concentration (P < 0.001) in the peripheral blood. Similar correlations were found also in intestinal mucosa: MnSOD: P < 0.05; CuZnSOD: P < 0.05; LOOH: P < 0.001. Our results show that oxidant/antioxidant balance is disturbed in CD patients with mucosal lesions. An increase in SOD activity as a consequence of oxidant pressure is not enough to maintain the normal level of free radicals, which leads to enhanced lipid peroxidation. These processes persist even in some patients on a long-term GFD.

Chemical composition and spasmolytic activity of Cymbopogon schoenanthus (L.) Spreng. (Poaceae) essential oil from Sudan

The plants of genus Cymbopogon are well known for their use in traditional medicine and for their high content of essential oils that are widely used as flavoring agents, fragrances, cosmetics, and pharmaceuticals. Essential oils isolated from the dried stems and inflorescence of cultivated Cymbopogon schoenanthus (L.) Spreng. (Poaceae) collected from Khartoum state (Sudan) in two periods were studied. The results of chemical analysis of C. schoenanthus revealed that this plant is rich in essential oil which is a valuable source of the commercially important monoterpene, piperitone (47.7-71.5%). Other abundant constituents of the oils were intermedeol (6.1-17.3%), δ -2-carene (4.5-10.0%) and elemol (2.7-9.0%). The essential oil from the inflorescence was tested for spasmolytic activity using three different experimental models: against spontaneous contractions, contractions induced with acetylcholine and contractions induced with potassium chloride. The oil exhibited strong, significant and dose-dependent spasmolytic activity, indicating the possibility for further investigations of this essential oil for its medicinal purposes or application in food industry. https://doi.org/10.2298/ABS160506113P Received: May 6, 2016; Revised: July 6, 2016; Accepted: July 6, 2016; Published online: November 11, 2016 How to cite: Pavlovic I, Omar E, Drobac M, Radenkovic M, Brankovic S, Kovacevic N. Chemical composition and spasmolytic activity of Cymbopogon schoenanthus (L.) Spreng. (Poaceae) essential oil from Sudan. Arch Biol Sci. 2017;69(3):409-15.

Increased Activity of Hippocampal Antioxidant Enzymes as an Important Adaptive Phenomenon of the Antioxidant Defense System in Chronically Stressed Rats

Abstract This study examined the effects of chronic restraint stress (CRS: 2 hours × 14 days) on gene expression of three antioxidant enzymes, copper, zinc superoxide dismutase (SOD 1), manganese superoxide dismutase (SOD 2) and catalase (CAT) in the rat hippocampus. Also, we examined changes in the activities of SOD 1, SOD 2 and CAT in the hippocampus of chronically stressed rats. Investigated parameters were quantifi ed by using real-time RT-PCR, Western blot analysis and assay of enzymatic activity. We found that CRS did not change mRNA and protein levels of SOD 1 and CAT, but increased mRNA and protein levels of SOD 2. However, CRS treatment increased the enzyme activities of SOD 1, SOD 2 and CAT. Our fi ndings indicate that the increased activity of antioxidant enzymes (SOD 1, SOD 2 and CAT) in the hippocampus may be an important adaptive phenomenon of the antioxidant defense system in chronically stressed rats.

Acute Effects of Smoking on Heart Rate and Peripheral Resistance in Young Smokers

Summary Smoking has many harmful effects on human body. It is well known that smoking is one of the most important risk factors for cardiovascular diseases. Though the ingredients from cigarette smoke stimulate the release of vasoconstrictor and reduce the release of vasodilator substances, the goal of this study was to investigate the acute effects of smoking on arterial blood pressure, heart rate and blood vessel flow. The investigation was conducted on forty healthy volunteers divided into two groups of twenty non-smokers (control group), and twenty smokers (experimental group). The group of smokers was examined before and after smoking four cigarettes during one hour. Blood pressure and electrocardiograph (ECG) were measured by common methods. Brecht’s and Boucke’s methods of plethysmography were used to evaluate the peripheral circulation. There were no differences in measured systolic and diastolic blood pressure and recorded ECG between non-smokers and smokers, neither between smokers before and after smoking. However, heart rate was increased by 29.57% after smoking in comparison to the value before smoking. Evaluation of plethysmographic parameters showed that amplitude was significantly decreased. Parameters of peripheral resistance were very increased: ductility was decreased by over 20%. It can be concluded that acute smoking does not significantly affect the level of blood pressure, although it increases heart rate. The shape of plethysmograms showed decreased systolic filling and decreased diameters of blood vessels. Sažetak Pušenje ima mnogo štetnih efekata na ljudski organizam. Dobro je poznato da pušenje predstavlja jedan od najvažnijih faktora rizika za pojavu kardiovaskularnih bolesti. Sastojci dima cigarete stimulišu oslobađanje vazokonstriktivnih, a inhibišu oslobađanje vazodilatatornih supstanci. Zato smo kao cilj našeg rada postavili praćenje akutnih efekata pušenja cigareta na arterijski krvni peitisak, srčanu frekvenciju i periferni protok. Istraživanje je vršeno na grupi nepušača (kontrolna grupa) i grupi pušača (eksperimentalna grupa). Klasičnim metodama su registrovani krvni pritisak i elektrokardiogram (EKG) kod nepušača, kao i kod pušača pre i nakon pušenja četiri cigarete u vremenskom razmaku od jednog sata. Periferna cirkulacija je procenjivana upotrebom pletizmografske metode Brecht and Boucke. Naši rezultati pokazuju da nije bilo razlika u vrednostima sistolnog i dijastolnog krvnog pritiska, kao ni u EKG nalazima između nepušača i pušača. Pomenuti parametri ostaju slični i kod pušača nakon pušenja. Međutim, srčana frekvencija se nakon pušenja značajno povećava, čak za jednu trećinu (29,57%), u poređenju sa vrednostima pre pušenja. Procena pletizmograma pokazuje da se njegova amplituda smanjuje, a da se vrednosti parametara koji ukazuju na periferni otpor značajno povećavaju. Rastegljivost krvnih sudova (širina i površina pletizmograma) se smanjuje za oko 20%, što ukazuje na smanjenje srednje vrednosti dijametra krvnih sudova. Može se zaključiti da pušenje nema značajne akutne efekte na krvni pritisak, iako povećava srčanu frekvenciju. Pletizmogramski nalaz ukazuje na smanjenje sistolnog volumena i dijametra krvnih sudova.