A Study On The Antibacterial,Antioxidant,Anti-diabetic And Anticarcinogenic Activities Of Bioactive Compounds From Some Aromatic Medical Plants

Pulicaria inuloides is one of the species of genus “Pulicaria” and family “Asteraceae”.It is widely used as an anticancer,antidiabetes,rheumatism,antibacterial anticoagulant agent,as well as a folk medicine for the treatment of gout,cystitis,nephritis,dysmenorrhea,tooth pain,headache.It is rich in alkaloids,volatile oils,terpenoids,phenolic acids,flavonoids and hydrolysable tannins.In the first phase: Isolation and identification of bioactive compounds was done from essential oil obtained from medical aromatic plants of Pulicaria inuloides-Astereceae and Ocimum forskolei-Lamaiceae leaves.Then evaluation of antioxidant,total phenols,and antimicrobiological activities were done.The results showed,P.inuloides had higher total phenol content than O.forskolei(144 ± 5.32 vs.54.6 ± 30 mg GAE/g extract),higher antioxidant activity(92.92 ± 0.10 % vs.26.76 ± 0.11 % scavenging activity;IC50,4.5 ± 0.05 vs.73.03 ± 0.05)and ?-carotene bleaching(90.77 ± 0.21 % vs.41.03 ± 6.35 % inhibition).P.inuloides essential oil inhibited all tested microorganisms except Salmonella typhimurium and Shigella dysenteriae with a minimum inhibitory concentration(MIC)of 3.0 ?g/m L against Escherichia coli.O.forskolei essential oil inhibited only Candida albicans.The second phase: Volatile compounds of P.inuloides and Ocimum frosklei roots were identified by gas chromatography–mass spectrometry.Results showed that P.inuloides had a higher total phenol content than O.forskolei(55.4 ± 0.1 vs.35.3 ± 0.2 mg GAE/g extract),lower antioxidant activity(43.97 ± 0.18 % vs.23.07 ± 0.06 % scavenging activity;IC50,31.18 ± 0.01 vs.80.01± 0.03)and ?-carotene bleaching(39.9±0.7% vs.25.3±0.3 % inhibition).P.inuloides essential oil inhibited all tested microorganisms except Salmonella typhimurium,Shigella dysenteriae and E.coli.In the third phase: Effect solvent type on extraction of polyphenol,tannins,anthocyanin and flavonoid contnets in Pulicaria inuloides(Arecaceae),and its antioxidant and antibacterial activities were studied.The extraction procedures were performed separately in an ultrasonic bath for 30 min(70 °C for ethanol and 60 °C for methanol,and diethyl ether)at agitation speed of(frequency)50Hz and(Speed)70 rpm,respectively.Results showed that total polyphenol content(TPC)total flavonoid(TFC),total tannins content(TTC)and antioxidant capacity were maximized using methanol.Methanol and ethanol extracts of P.inuloides showed lower antimicrobial activity at all the chosen microorganisms.The diethyl ether demonstrated noantibacterial activity against all tested bacteria.Fourth: GC-MS chromatogram of the ethanolic leaf extract of Pulicaria inuloides showed 10 peaks indicating the presence of ten compounds.Fifth phase of study: Isolation and identification of phenolic compounds from P.inuloides,its antioxidant and antibacterial activities and tocopherols were evaluated.The phenolic compounds are isolated by macroporous resin column chromatography,and then identified through HPLC.Thirteen fatty acids were determined;six were unsaturated;linoleic acid was found in the highest content(5.74%),followed by oleic acid acid(2.37%)and 6,9,12-Octadecatrienoic acid(1.61%).?-tocopherol was the highest content and was about 52.76% of the total tocopherols,followed by ?-tocopherol and ?-tocopherol(29.83% and 17.41%,respectively).In addition,highest level of K,Mg,Na,Fe and Ca of 159.5,29.5,14.2,13.875 and 5.225 mg/100 g were found in P.inuloides.Transmission Electron Microscope,Scanning electron microscopy(TEM,SEM-EDX)revealed interior structure of P.inuloides.Sixth: The essential oil of P.inouloides leaves was applied on sea food juvenile kachlan(Trachinotus ovatus,Linnaeus).Results showed that P.inuloides essential oil inhibited all tested microorganisms with a minimum inhibitory concentration(MIC)of 2.0 ?g/m L against Escherichia coli.The present study evaluated the impact of Pi EO on L.monocytogenes,E.coli,S.aureus and tap water(control)inoculated in fish fillets.Results indicated that Pi EO eliminated most of L.monocytogenes,E.coli,S.aureus and SPC immediately after treatment at time zero(25 °C)and during storage at 10 °C for 12 days,and it increased the shelf-life of seafood during storage at 10 °C compared with the tap water;whereas SPC counts were > 7 log cfu/g at the end of storage.The sensory evaluation of fillet fish treated with Pulicaria inuloides essential oil was acceptable by consumers.Fish purchased from supermarkets in Wuxi Jiangsu,China,were maintained with good hygiene practices and health conditions;whereas,fish purchased from local markets were not.The next phase: The leaves of P.inuloides essential oil are considered useful for the treatment of diabetes mellitus.The results of phytochemical analysis of the essential oil revealed the presence of alkaloids,steroids,saponins,glycosides,raducing sugar and terpenoids.At a concentration of 250 ?g/m L;the leaves of P.inuloides essential oil exhibited ?-amylase and ?-glucosides inhibitory activity of 85.50,86.52 %,with IC50 values of 140.49,128.33 ?g/m L,respectively.Leaves of pulicaria inuloides essential oil decreased fasting blood glucose levels in diabetic rats at the dose level(400 mg per kg body weight).Leaves of pulicaria inuloides essential oil at day 21 reduced the blood glucose to levels that were not statistically significant(P<0.05)compared to the diabetes drug group.Essential of P.inuloides leaves oil reduced total cholesterol,triglycerides and LDL but increased the HDL-cholesterol levels in STZ-diabetic rats.During the treatment for 21-days,weight remained stable in treated rats.Levels of liver function marker enzymes ALT,AST and ALP were significantly elevated in STZ-induced diabetes.The rats treated with Pulicaria inuloides(leaves)showed significant(p<0.01)reduction in the elevated levels of these enzymes.Kidney function markers like creatinine and urea were elevated in the STZ-induced diabetic rats when compared with the normal rats.Essential oil of Pulicaria inuloides leaves reduced in both levels.Also,the lower level of total protein in diabetic rats was significantly(p<0.01)elevated after 21 days treatment: these results thus indicated that diabetic rats did suffer renal dysfunction Histopathological studies of a section of the liver showed that essential oil of P.inuloides extensively improved islets cells as normal cells in compared with control and standard.The results of possible toxic effects of plant part did not show any visible signs of acute toxicity such as respiratory distress,coma or death.In the last part: The P.inuloides essential oil tested in our study were showed to have strong cytotoxic activity against MCF-7 cells at all times compared with control.The oil showed higher cytotoxic activity,with an IC50 = 12.9 and 3.00 ?l/m L,respectively.P.inuloides essential oil(Pi EO)inhibited all tested ListeriaMorphological analyses of MCF-7cells were carried out by light microscopy...