Chemical Analysis and Anticancer effects of Juniperus polycarpos and Oak Gall plants extracts

 

Raghad Dhyea Abdul Jalill

College of Science, Mustansiryiah University, Baghdad  Iraq.

 

ABSTRACT:

The seeds of Juniperus polycarpos and Oak Gall (distortion of leave buds of Quercus infectoria)were investigated for chemical composition by Gas Chromatography Mass Spectrum (GC-MS)and studied their anticancer activity. Sixty-five different compounds recognized in crudefruits extract of J.polycarpos. The major three components were: 2(10)-Pinene, (1S,5S)-(-)- (17.78%), 2-Hexanol, 2-methyl- (21.14%) and 2,4-Decadienal (8.72%).They were presence of thirty-four compounds in the crud extract of Oak Gall. The highest strong peak area (69.4%) was obtained by 2-Hexanol, 2-methyl- fallowed by 2,4-Decadienal (6.79%) and Eucalyptol (5.17%). There are some similar compounds in these plants. 2-Octenal, (E)-; 2,4-Decadienal and Hexanoic acid is exactly the same compounds found in alcohol extracts of J.polycarpos andOak Gall. Moreover, 2-Heptenal, (Z)-; 2-Hexanol, 2-methyl-; Copaene; 2-Decenal, (Z)-; .alpha.-Terpieol and  2,4-Decadienal, (E,E)- were found in J.polycarpos and in Oak Galls All of concentrations of crud plant extracts of J.polycarpos andOak Gall had high reductions to mice mammary carcinoma cell line 2003 AMN3 cancer cell line, their half maximal inhibitory concentration (IC₅₀) 2 microgram per milliliterand 0.2 milligram per milliliter respectively. Less reductions found on recombinant mouse epithelial cell line (L₂₀B) cell line. IC₅₀ of J.polycarposwere: 0.2 mg/ml. 

 

 

 

INTRODUCTION:

J. polycarpos K. Koch/ family: Cupressaceae, distributed in: north west of Iran, (1)Turkia and north of Iraq (2). It is very a variable species, often include as a subspecies or variety of Juniperus excelsa(3) their DNA analysis is certainly different(4-5).

 

Chemical analyses found presence high cedrol(6) and zero cedrol chemotypes in addition to ascorbic acid, 57.84 mg %, proline 34 µg/g, soluble phenols, 6.83 mg/g besides anthocyanins, 0.411 mg/g, soluble carbohydrates 2.3 mg/g and proteins 4.88 % in leaves which has antioxidant activity 77 % (7). Pyrogallol (1.6 L^-1) and quercetin (0.129 L^-1) were also present (8).

 

 

Oils of leaves contains: Shyobunol; α -eudesmols; germacrene B; (E)-caryphyllene; β-eudesmols; elemol; α-murrolol; epi-α-muurolol; epi-α-cadinol; junenol, α-muurolene; plus (4Z)-decen-1-ol; germacrene D-4-ol and γ-cadinene,(9). Moreover, oil of leaves of J. polycarpos var. polycarpos consist of: cedrol (13.1 %), α-pinene (32.3 %), myrcene, (3 %), β-elemene (3.7 %), limonene (2.8 %) and β-bisabolene (2.9 %); although, the oils of fruits contained: germacrene B (3.2 %) (10), terpine-4-ol, limonene, sabinene, γ-terpinene and δ-cadinene, (11). The fruits of J. polycarpus var. seravschanica contain abietanes, eudesmanes and podocarpanes, (12).

 

Common classic uses of J. polycarpos is: common cold, urinary infections urticaria, rheumatic arthritis, hemorrhage, dysentery and remove the pain of menstrual (13). Recently, several researches approved their antimicrobial, antifungal and antioxidant activity (14) with protective effects on leucocytes (8), and anticancer, (15).

 

Oak Gall are a distortion of base of leaves of Quercus infectoria (Familly: Fagaceae) caused by gall Oak wasps, Andricus spp. which is the most spread insects on Oak trees in northerly of Iraq, (16-17). The galls contain: gallic acid, tannin, gallo tannic acid (50-70) %, rubric acid, ellagic acid, nyctanthic acid, besides sugars, starch, anthocyanins in addition to an essential oil. Galls contain: beta-sitosterol, amentoflavone, isocryptomerin, and hexamethyl ether (18). Morover, caffeic acid, chlorogenicn acid in addition to cinnamic acid were also found (19).

 

Traditionally, it is used for diseases of the gums and oral, haemorrhoids, breast and vaginal firming creams and to treat prolapus of rectum, (18).

 

Recently, some documented possess it as: antibacterial, (20-21), astringent, antiviral, Antiprotozoal, (22), antitremorine, local anaesthetic, antidiabetic, antifungal, larvicidal, and anti-inflammatory activities, (23), Antioxidant, (24). In addition, Gall extract induced activity and proliferation of hFOB cell line (human osteoblast), (25). The aqueous, ethanol and methanol Gall extracts induced anticancer activity in (HeLa) and (Caov-3) cancer cells, (26).

 

The aims of the present work were determining some elements and qualitative/ quantitative chemical analysis of Oak Gall and cones of J. polycarpos extracts using ABB and GC/MS techniques and if crude extracts could inhibit cancer activity.

 

MATERIALS AND METHODS:

Preparation and extraction of plants:

The Oak Gall (distortion of base of leaves of Quercus infectoria) and cones of Juniperuspolycarpos, plants obtained from Iraq Medical Herbarium/ Ministry of Health/ Baghdad / Iraq. J. polycarpos classify according to (5). Dried at 38 Cº and ground by a grinder. Fifty grams of powdered material were extracting using ethanol (250 mL, 70 %) by a Soxhlet extractor at forty centigrade for three hours. (27). Whatman No.1 and 0.22 μ micro filters were used to filtere and sterilized extracts.

 

Chemical analysis:

Atomic Absorption Spectrometer (AAB):

Atomic Absorption Spectrometer (from Analytik Jena; Nove AA-350) was used to determine the presence of the following elements in Oak Gall and J. polycarpos: Cu, Zn, Cd, Fe, Mn, K, in addition to: Co, Ti, P and N. The samples prepared according to Kodama in (1963)(28). The levels of Nitrogen and total proteins were determined using the Macro Kjeldahl method (29).

 

Gas Chromatography Mass Spectrum (GC-MS) tests:

GC-MS analyses had done through Shimadzu GC-2010 combine with Shimadzu GCMS-Q2010 Ultra. Capillary column was InertCap (Sciences/ Japan), their dominations were: 0.25 µm; 30 m; 0.25 mm. Carrier-gas was helium. Constant flowrate was 1 ml/min, auto injector was AOC-20i, Shimadzu, Split Ratio was 2.0. Program's oven temperature illustrated in table (1).

 

Identifying the components approved through matching the retention times together with the data of mass spectral of components which found in extracts with those standard compounds in Standard Reference Databases which found in Mass Spectral Library (NIST08.LIB) complemented with NIST Mass Spectral Search Program (Ver. 2.0f). The matching target compounds were more than 95%.

 

Cytotoxicity on cell lines:

Cell culture:

Cell lines were got from the Iraqi Center for Cancer and Medical Genetic Research (ICCMGR) / Department of Experimental Therapy, Cell Bank Unites, they were: mice mammary carcinoma cell line 2003 (AMN₃) (passage number 210)(1), (30), and recombinant mouse epithelial cell line which expresses a receptor of human poliovirus on the cell surface (L₂₀B) (passage number CD/155), (31). Preparation of the cell lines were carried out according to ICCMGR. They maintained using RPMI₁₆₄₀ (USbiological) complemented with 15% calf bovine-serum (Gibco, USA), 100 μg/ml streptomycin and 100 units/ml penicillin. The cells were culturing in Flat bottomed 96-well polystyrene and incubated at 37 °C provide by CO₂ (5 %). The viable cell count had done using the trypan blue technique according to (32). Populations of cells were 1.5*104 per cells.  

 

Cytotoxic activity:

Cytotoxicity test was depending on the method of Department of Experimental Therapy of ICCMGR center with a few changes. The media removed when cells reach (90-95) % monolayer confluence cells. PBS (Sigma) was used to wash cells then the media (RPMI 1640) without serum (0.2 ml per well) were added. Separately, serial dilutions of each crude plant extract, dissolved in PBS (which were diluted with media (RPMI 1640) without serum were added to flat bottomed 96-well polystyrene plates. There were a negative control (PBS) and six-fold replicates for each treatment. Cells incubated at thirty-seven centigrade supplemented with CO₂ (5%). After 24 hours, cells washed with PBS twicely. Fifty-micro letter of the Neutral Red solution (Sigma, U.S.A; 50 μg/ml dissolved in PBS), used to stained cells for two hours at thirty-seven centigrade incubation.  Wells washed three times with PBS. Fifty microliters of solution (PBS and absolute ethanol 1:1 V/V) added to each well. A Micro-ELISA was used to read the optical density. Percentage of cell viabilities)(33) and inhibiting rates (34)were calculated.

 

RESULTS:

Chemical analysis

AAB:

The levels of Mn, Zn, Fe, Cd, K in addition to Cu, Co, Ti and N showed in Table (2). Phos­phorus was not found. The level of total proteins was 2.4125 % and 2 % respectively.

 

GC-MS analyses:

J. polycarpos: Seventy different compounds recognized in Jpolycarpos (Table 3). The major three components present were: 2(10)-Pinene, (1S,5S)-(-)- (17.78%),  2-Hexanol, 2-methyl- (21.14%) and 2,4-Decadienal (8.72%).

 

Oak Gall:

table (4) showed the presence of thirty-four compounds in the crud extract of Oak Gall. The highest strong peak area (69.4%) was obtained by 2-Hexanol, 2-methyl- fallowed by 2,4-Decadienal (6.79%) and Eucalyptol (5.17%). Table (1 and 2) showed retention time, chemical formula, molecular weight and mass spectraof all compounds found in J. polycarpos and Oak Gall extracts. Retention time, chemical formula, molecular weight and mass Spectra ofthese two extracts were present in (table 3, 4) respectively and chromatogram were present in (Figure 1, 2) respectively.

 

Table (1): Oven temperature program of GC-MS.

P. E	I. V. (µl)	R.	T. (°C)	H. T. (min)
J.polycarpos	1	-	70	3
		20	150	1.5
		6	220	5
Oak Gall	1	-	70	3
		15	120	2
		10	200	8

PE: plants extracts; I.V.: injection volume (µl); R: rate; T: temperature (°C); H.T.: hold Time (min).

 

Table 2: The level (%) of some elements and total proteins in Oak Gall and J. polycarpos.

Element	%
	J. polycarpos	Oak Gall
Fe	1051*10-5	2895 *10-6
Zn	104 *10-5	18 *10-7
Cu	5 *10-5	2002*10-7
MN	75 *10-5	1826*10-7
K	413 *10-3	848 *10-3
Cd	25 *10-5	289*10-7
Co	22 *10-5	178 *10-7
Ti	23 *10-3	15 *10-3
P	0	0
N	386*10-3	32*10-2
protein	2.413	2

Elem.: element; %: Percentage; Zn: Zinc; Fe: Iron; Mn: Manga­nese; Cu: Copper; K: Potas­sium; Cd: Cad­mium; Co: Cobalt; Ti: Tita­nium; P: Phos­phorus; N: Nitro­gen; Prot.: Crude protein.

 

Table (3): Chemical compositions of extract of J.polycarpos

P	R.T.	Area %	Name	Ch.F.	M.W.	M.S
1	3.6	0.22	2,2-Dimethyl-3-methylenebicyclo[2.2.1]heptane	C10H16	136
2	3.68	0.87	Hexanal	C6H12O	100
3	4.1	0.12	Cyclopentene, 3-isopropenyl-5,5-dimethyl-	C10H16	136
4	4.52	17.78	2(10)-Pinene, (1S,5S)-(-)-	C10H16	136
5	4.94	2.83	Cyclobutane, 1,2-bis(1-methylethenyl)-, trans-	C10H16	136
6	5.05	0.18	.beta.-Phellandrene	C10H16	136
7	5.27	0.04	Furan, 2-pentyl-	C9H14O	138
8	5.41	0.02	1-Pentanol	C5H12O	88
9	5.45	0.04	p-Menth-1-en-4-ol, acetate	C12H20O2	196
10	5.73	0.73	o-Cymene	C10H14	134
11	6.29	1.16	2-Heptenal, (Z)-	C7H12O	112
12	6.66	21.14	2-Hexanol, 2-methyl-	C7H16O	116
13	6.86	0.7	Pinane, 2,3-epoxy-	C10H16O	152
14	7.15	1.24	9,10-Diazatricyclo[4.4.0.0(2,8)]dec-9-ene	C8H12N2	136
15	7.26	0.11	8,11,14-Eicosatrienoic acid, methyl ester, (Z,Z,Z)-	C21H36O2	320
16	7.35	0.19	2-Octenal, (E)-	C8H14O	126
17	7.42	0.1	Acetic acid	C2H4O2	60
18	7.49	0.12	1-Octen-3-ol	C8H16O	128
19	7.73	0.81	.alpha.-Cubebene	C15H24	204
20	8	0.25	Cyclohexene, 4-isopropenyl-1-methoxymethoxymethyl-	C12H20O2	196
21	8.1	0.63	Copaene	C15H24	204
22	8.33	0.13	DL-Camphor	C10H16O	152
23	8.47	0.46	allo-Ocimenol	C10H18O	154
24	9	0.48	1,7,7-Trimethylbicyclo[2.2.1]hept-2-yl acetate	C12H20O2	196
25	9.12	0.11	Cyclohexane, 2,4-diisopropenyl-1-methyl-1-vinyl-, (1S,2R,4R)- (-)-	C15H24	204
26	9.18	0.07	p-Menth-1-en-4-ol, (R)-(-)-	C10H18O	154
27	9.26	0.42	Farnesene epoxide, E-	C15H24O	220
28	9.53	0.14	(1R)-(-)-Myrtenal	C10H14O	150
29	9.64	0.42	2-Decenal, (Z)-	C10H18O	154
30	9.78	0.41	2(10)-Pinen-3-ol, (1S,3R,5S)-(-)-	C10H16O	152
31	9.89	0.24	(Z)-.beta.-Farnesene	C15H24	204
32	10.01	0.22	2-Pinen-4-ol	C10H16O	152
33	10.1	0.16	1,4,7,-Cycloundecatriene, 1,5,9,9-tetramethyl-, Z,Z,Z-	C15H24	204
34	10.2	0.12	.alpha.-Terpieol	C10H18O	154
35	10.28	1.48	.gamma.-Muurolene	C15H24	204
36	10.47	0.36	l-Verbenone	C10H14O	150
37	10.6	0.15	(Z,Z)-.alpha.-Farnesene	C15H24	204
38	10.69	1.11	Guaia-1(5),11-diene	C15H24	204
39	10.94	0.11	2-Decenal, (E)-	C10H18O	154
40	11.07	6.22	2,4-Decadienal, (E,E)-	C10H16O	152
41	11.27	0.04	.alpha.-Curcumene	C15H22	202
42	11.68	8.72	2,4-Decadienal	C10H16O	152
43	11.96	0.96	Hexanoic acid	C6H12O2	116
44	12.1	0.16	Carbonic acid, 4-isopropylphenyl propargyl ester	C13H14O3	218
45	12.42	0.16	N-[4-(4-Chlorophenyl)isothiazol-5-yl)-1-methylpiperidin-2-imine	C15H16ClN3S	305
46	13.26	0.52	Acetic acid, 6,6-dimethyl-2-methylene-7-(3-oxobutylidene)oxepan-3-ylmethyl ester	C16H24O4	280
47	13.51	0.38	Selina-6-en-4-ol	C15H26O	222
48	13.64	0.14	3-Heptanol, 6-methyl-	C8H18O	130
49	14.12	4.54	Caryophyllene oxide	C15H24O	220
50	14.37	0.28	Lanceol, cis	C15H24O	220
51	14.63	0.21	2,2-Dimethyl-3-[3,7,12-trimethyl-14-(1,4,4-trimethylcyclohex-2-enyl)tetradeca-3,7,11-trienyl]oxirane	C30H50O	426
52	15.07	2.51	1,5,5,8-Tetramethyl-12-oxabicyclo[9.1.0]dodeca-3,7-diene	C15H24O	220
53	16.06	1.08	Longipinocarveol, trans-	C15H24O	220
54	16.93	0.4	γ-Eudesmol	C15H24O	220
55	17.52	0.46	(6E,8E,10E)-2,6,11,15-Tetramethyl-2,6,8,10,14-hexadecapentaene	C20H32	272
56	17.7	0.5	Hexadecanoic acid, methyl ester	C17H34O2	270
57	17.78	0.7	Tricyclo[5.2.2.0(1,6)]undecan-3-ol, 2-methylene-6,8,8-trimethyl-	C15H24O	220
58	18.13	0.84	2-Pentene, 1-ethoxy-4,4-dimethyl-	C9H18O	142
59	18.74	2.96	6-Isopropenyl-4,8a-dimethyl-1,2,3,5,6,7,8,8a-octahydro-naphthalen-2-ol	C15H24O	220
60	20.29	0.34	Kaurene	C20H32	272
61	20.82	0.18	Heptadecanoic acid, 15-methyl-, methyl ester	C19H38O2	298
62	21.15	1.49	9-Octadecenoic acid (Z)-, methyl ester	C19H36O2	296
63	21.94	2.59	Methyl linoleate	C19H34O2	294
64	22.27	0.92	.gamma.-Thionodecalactone	C10H18OS	186
65	22.46	7.09	Cerasynt	C22H44O4	372
		100

R.T: Retention time (min); Ch.F: Chemical Formula;M.W: Molecular weight; M.S: Mass Spectra.

 

 

Fig. (1). Chromatogram ofJ.polycarpos

 

Table (4): Chemical compositions of extract ofOak Gall

P	R.T.	Area%	Name	Ch.F.	M.W.	M.S.
1	3.7	0.53	Hexanal	C6H12O	100
2	5.07	0.56	m-Mentha-6,8-diene, (R)-(+)-	C10H16	136
3	5.2	5.17	Eucalyptol	C10H18O	154
4	5.66	0.23	1,4-Cyclohexadiene, 1-methyl-4-(1-methylethyl)-	C10H16	136
5	5.96	0.18	1,3,8-p-Menthatriene	C10H14	134
6	6.14	0.11	(+)-4-Carene	C10H16	C10H16
7	6.62	1.01	2-Heptenal, (Z)-	C7H12O	112
8	7.12	69.45	2-Hexanol, 2-methyl-	C7H16O	116
9	8.16	0.19	2-Octenal, (E)-	C8H14O	126
10	9.35	0.09	Copaene	C15H24	204
11	9.88	0.17	2-Nonenal, (E)-	C9H16O	140
12	9.96	0.07	1H-Cycloprop[e]azulene, 1a.beta.,2,3,4,4a.alpha.,5,6,7b.beta.-	C15H24	204
13	10.44	0.08	2(10)-Pinen-3-one, (.+/-.)-	C10H14O	150
14	10.92	0.46	p-Menth-1-en-4-o	C10H18O	154
15	11	0.16	4,8,8-Trimethyl-2-methylene-4-vinylbicyclo[5.2.0]nonane	C15H24	204
16	11.16	1.3	1H-Cycloprop[e]azulene, decahydro-1,1,7-trimethyl-4-methylene	C15H24	204
17	11.57	0.73	2-Decenal, (Z)-	C10H18O	154
18	11.73	0.85	(+)-Sativen $$ 1,4-Methanoindan, hexahydro-7-isopropyl	C15H24	204
19	12.17	0.36	2-Isopropenyl-5-methylhex-4-enal	C10H16O	152
20	12.31	0.78	p-Menth-1-en-8-ol, (S)-(-)-	C10H18O	154
21	13.28	4.29	2,4-Decadienal, (E,E)-	C10H16O	152
22	13.9	6.79	2,4-Decadienal	C10H16O	152
23	14.11	0.13	Anisole, p-propenyl-	C10H12O	148
24	14.25	0.3	Hexanoic acid	C6H12O2	116
25	14.34	0.07	Adamantane, 1-thiocyanatomethyl-	C12H17NS	207
26	15.7	0.6	Ethanol, pentamethyl-	C7H16O	116
27	16.66	0.17	2-Propenal, 3-phenyl-	C9H8O	132
28	16.84	0.93	Octanoic Acid	C8H16O2	144
29	17.23	0.43	Humulane-1,6-dien-3-ol	C15H26O	222
30	18.29	0.87	Eugenol	C10H12O2	164
31	19.67	0.42	2,2,3,3-Tetraethyloxirane	C10H20O	156
32	19.85	0.67	Pyrrole-2-carboxylic acid, 4-(1-chlorodec-1-enyl)-3,5-dimethyl-, ethyl ester	C19H30ClNO2	339
33	22.53	1.42	Kaur-16-ene	C20H32	272
34	23.91	0.45	Methyl 11-octadecenoate	C19H36O2	296
		100

R.T: Retention time (min); Ch.F: Chemical Formula;M.W: Molecular weight; M.S.: Mass Spectra

 

 

Fig. (2). Chromatogram of Oak Gall .

 

Fig. (3): Cytotoxicity assay of J. polycarpos on different cell lines, CT-: negative control, on % I.R.: percentage of inhibition rate.

 

 

Fig. (4): Cytotoxicity assay of Oak Gall on different cell lines, CT-: negative control, on % I.R.: percentage of inhibition rate.

 

Cytotoxicity on cell lines:

The results of J. polycarpos extract present reduction in the cell viability of AMN3 cancer cell line by all concentrations with IC₅₀ 2 µg/ml while they were less reducer to L₂₀B cell line, IC₅₀ was 0.2 mg/ml, (Fig. 3). Different concentrations of Oak Gall extract reduced AMN3 cancer cell line, the IC₅₀ was 0.2 mg/ml. L₂₀B was less affected by (0.2 µg/ml-2 mg/ml), the higher negative effect on L₂₀B was 45%, which was the inhibition rate using 2 mg/ml, while there was a positive effect (over growth) at 200 mg/ml, there was no IC₅₀, (Fig. 4).

 

DISCUSSION:

In current work, seventy different compounds were identified in crude extract of J. polycarpos. They were presence of thirty-four compounds in the crude extract of Oak Gall. There are some similar compounds in these plants. 2-Octenal, (E)-; 2,4-Decadienal and Hexanoic acid is exactly the same compounds found in alcohol extracts of J. polycarpos and Oak Gall. Moreover, 2-Heptenal, (Z)-; 2-Hexanol, 2-methyl-; Copaene; 2-Decenal, (Z)-; .alpha.-Terpieol and  2,4-Decadienal, (E,E)- were found in J. polycarpos and in Oak Galls. In this study, all concentrations of crud plant extracts of J. polycarpos and Oak Gall had high reductions to AMN3 cancer cell line.

 

The anti-cancer activity of J.polycarpos plant extracts is may cause by some compounds which were found in this study such as: 2,4-decadienal (8.72 %), caryophyllene oxide (4.54%), and Copaene (0.63%).

 

2,4-decadienal (which appear at 8.72% in the current study) could inhibit cell viability of HEL-TIB 180 cell line (human erythroleukemia) with fragmentation of DNA,)(35).

 

Caryophyllene oxide (4.54% in the current study) caused tumorigenesis, which could be prevent and treat cancer which acts as mitogen activated protein kinases "MAPK" and "PI3K/AKT/mTOR/S6K" activation pathways in breast and prostate human cancer cells respectively. β-caryophyllene oxide inhibited activation of intracellular signaling pathway PI3K/AKT/mTOR which regulate cell cycle in addition to reduction of: P₃₈ and JNK in addition to ERK in breast tumor cells, (36).

Another study found that caryophyllene oxide activate apoptosis in neuroblastoma cells and lymphoma through modifying the up-stream target of 15-LOX subsequently by regulation of antiapoptotic and proapoptotic genes (37).

 

Copaene (this compound found at 0.63 % and 0.09 % in both J. polycarpos and Oak Gall in this study) caused moderate cytotoxic effects against neuron and neuroblastoma cell lines, (38).

 

Many species related to J. polycarpos species have anticancer activity. The J. sabinaand J. foetidissimareduced cell viability of some cancer cell lines, (39). Moreover, ethanol extracts of branchlets of J. excelsa subsp. polycarpos (which is J. polycarpos according to recent classification, and J. foetidissima demonstrate an inhibitory effect against KB, hella and MDA-MB-468 cell line however, J. excelsa subsp. excels indicated reduction in KB cells, (15). In addition, the Hella cell line was not affected by J. sabina, (15).

 

The anti-cancer activity of Oak Gall extracts is possibly due to some compounds which was found in current study such as: Eucalyptol (5.17 %), Eugenol (0.87) and Kaur-16-ene (1.42%). Many documentations approved this meaning. Eucalyptol could suppress production of various factors in cells: leukotriene, alpha-TNF, thromboxan and interleukin-1beta, (40), and inhibited human colorectal cancer proliferation through apoptosis (41).

Eugenol (0.87 % in Oak Gall) caused apoptosis in various human cancer cells, such as: breast cancer and colon, (42); (43), in addition to: melanoma, leukemia, gastric cancer, osteosarcoma and skin tumor, (Jaganathan and Supriyanto 2012)(44), prostate tumor cells and oral squamous carcinoma cells, (45).

 

Various Kaur-16-ene (synonym is Ent-kaurene found in Oak Gall 1.42%) diterpenes compounds is a wide spectrum anticancer effects against different cell lines of human which could inhibit cell-cycle progression and induced programmed cell death in: HL-60 cell line (promyelocytic leukemia of human), (46), 22Rv1 and LNCaP cell lines (human prostate), HT29, HCT116, SW480 and SW620 cell lines (colon cancer) and MCF-7 cell lines (breast tumor) (47).

 

Recently, some documented possess Oak Gall extract as reduced proliferations and activation of fetal osteoblast cell line of human (hFOB 1.19), (25) and  as an anticancer agent againstHeLa/ ovarian cell line and Caov-3 cell line and on non-malignant cell line /MDCK at seventy-two hours incubation time. The IC₅₀ was 2.82 μg/ml for HeLa cell line which treated with alkohol extract. Moreover, IC₅₀ was 6.50 μg/ml for Caov-3 cell line by aqueous extract while there was less toxic to MDCK normal cell line treated with ethanol extract (IC₅₀ was 74.99 μg/ml). Ethanol extracts loss their attachments to other cells with condensate the chromatin(26).

 

CONCLUSIONS AND RECOMMENDATIONS:

Sixty-five different compounds were identified in crude extract of J.polycarpos and thirty-four compounds in crude extract of Oak Gall. There are some similar compounds in these three plants. All concentrations of crud plant extracts of J. polycarpos and Oak Gall (AMN3 cell line) and these effects were less than the reduction on L₂₀B cell line. More studies about the toxicity of major compounds which found in these plants on different cell line could be benefits.

 

ACKNOWLEDGMENTS:

Many thankfulness to Mustansiriyah University (www. uomustansiriyah.edu.iq) Baghdad/ Iraq for their supports in this work.

 

FUNDING:

There are currently no Funding Sources.

 

CONFLICT OF INTEREST:

The authors declare that they have no conflict of interest.

This article does not contain any studies with human participants or animals performed by any of the authors.”

 

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Accepted on 15.02.2018           © RJPT All right reserved

Research J. Pharm. and Tech 2018; 11(6): 2372-2387.