Phytochemical Investigation and Anti-Fertility Activity of Lichen Parmelia Perlata

Abstract

The present work deals with the investigation of the petroleum ether extract of lichen Parmelia perlata. The study led to the isolation of compounds-(I) Stigma-4,22-diene-3-one, (II) Stigmasterol, (III) 5,4´-Dihydroxy-3,6,7,3´,5´pentamethoxy flavone, (IV) Benzyl-2,6-dimethoxy benzoate, (V) Benzyl-2,3,5,6-tetramethoxy benzoate. Compounds III-V have been isolated for the first time from this lichen. Also the anti-fertility activity of the crude extract was studied. Parmelia perlata exhibited anti-spermatogenic effect in treated rats. The pet-ether extract of Parmelia perlata decreased the fertility of male albino rats by 100%.There was a marked decrease in the number of primary and secondary spermatocytes and spermatids. A significant reduction was found in seminiferous tubular diameter and differential count of Leydig cells. Keywords-Parmelia perlata, 5,4´-dihydroxy-3,6,7,3´,5´pentamethoxy flavone, benzyl-2,6-dimethoxy benzoate, benzyl-2,3,5,6-tetramethoxy benzoate, anti-fertility, male albino rats.

Full text

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International Journal of Recent Research and Review, Vol. VIII, Issue 4, December 2015
ISSN 2277 – 8322
Phytochemical Investigation and Anti-Fertility Activity of Lichen
Parmelia Perlata
Kumud Tanwar1, Jaya Mathur2 and J.B.S.Kachhawa3
1Department of Chemistry, JNIT, Sitapura, Jaipu , Rajasthan (India)
2Department of Chemistry, Rajasthan University, Jaipur, Rajasthan (India)
3Department of Zoology, Rajasthan University, Jaipu , Rajasthan (India)
Email: tanwar_ kumud @ yahoo.co.in
Abstract- The present work deals with the investigation of
the petroleum ether extract of lichen Parmelia perlata. The
study led to the isolation of compounds- (I) Stigma-4,22-
diene-3-one, (II) Stigmasterol, (III) 5,4´-Dihydroxy-
3,6,7,3´,5´ pentamethoxy flavone, (IV) Benzyl-2,6-dimethoxy
benzoate, (V) Benzyl-2,3,5,6-tetramethoxy benzoate.
Compounds III-V have been isolated for the first time from
this lichen. Also the anti-fertility activity of the crude
extract was studied. Parmelia perlata exhibited anti-
spermatogenic effect in treated rats. The pet-ether extract
of Parmelia perlata decreased the fertility of male albino
rats by 100%.There was a marked decrease in the number
of primary and secondary spermatocytes and spermatids. A
significant reduction was found in seminiferous tubular
diameter and differential count of Leydig cells.
Keywords- Parmelia perlata, 5,4´-dihydroxy-3,6,7,3´,5´
pentamethoxy flavone, benzyl-2,6-dimethoxy benzoate,
benzyl-2,3,5,6-tetramethoxy benzoate, anti-fertility, male
albino rats.
I. INTRODUCTION
Parmelia perlata is a well known lichen of family
Parmeliaceae. A lichen is an association of an alga and
fungus, living together in a symbiotic relationship.
Parmelia perlata is commonly called Stone flower or
Chadila. In India it is mainly found in Himachal Pradesh
and West Bengal. It is used as food, fodder and medicine.
It is a good pain reliever and is used as a remedy for early
healing of wounds. It cures many skin diseases and is
considered to be an expectorant, astringent, resolvent,
laxative, carminative and aphrodisiac. It is also used in
treatment of fever, cough, dysentery and renal calculi.
This lichen exhibits antimicrobial [1-2], antiviral [3], anti-
tumor [4], antispasmodic [5] , antioxidant [6] and
antipyretic [7] activities. Its hepatoprotective action [8]
has also been reported. Phytochemical studies of
P.perlata have led to the isolation of various chemical
constituents such as atranorin, chloroatranarin, salazinic
acid [9], lecanoric acid [10], imbricaric acid [11],
lecanora. Two terpenes, parmelanostene and
permelabdone [12] and usnic acid [13] have also been
isolated from this lichen.
Encouraged by the presence of interesting chemical
compounds in this lichen, we examined its petroleum
ether extract. The study led to the isolation of five
compounds- (I) Stigma-4,22-diene-3-one, (II)
Stigmasterol, (III) 5,4´-Dihydroxy-3,6,7,3´,5´
pentamethoxy flavone, (IV) Benzyl-2,6-dimethoxy
benzoate, (V) Benzyl-2,3,5,6-tetramethoxy benzoate.
Compounds III-V has been isolated for the first time from
this lichen. In pursuing our interest in its medicinal and
bioactive properties, the anti-fertility activity of the
extract was also studied.
II. MATERIALS AND METHODS
A) Plant material
The plant material Parmelia perlata (lichen) was
collected from the hills of Himachal Pradesh (India).
Identification of the lichen was done with the help of
Department of Botany, University of Rajasthan, Jaipur,
India and a voucher specimen was deposited at RUBL
Herbarium, Jaipur (RUBL 3674).

31
B) General experimental procedures
Melting points were determined in soft glass capillaries in
an electrothermal melting point apparatus and are
uncorrected. TLC (both Qualitative and quantitative) was
performed on aluminium sheet Kieselgel 60 F254 (E.
Merck). For column chromatography silica gel (E. Merck,
60-120 mesh, 550 gm) was used. The IR spectra (with
KBr pellets, ν in cm-1) were recorded on FTIR
SHIMADZU 8400S spectrometer . The 1H and 13C NMR
spectra (δ in ppm, J in Hz, in CDCl3) were recorded at
300 MHz and 125 MHz on a Brucker NMR instrument,
respectively, using TMS as an internal standard. FAB
mass spectra were recorded on JEOL SX 102 /DA-6000
mass spectrometer using Argon/Xenon as FAB gas.
C) Extraction and isolation
Shade dried lichen was powdered and extracted with
petroleum ether on steam bath for 3 x 12 hrs. The crude
extract, after removal of solvent, was obtained as a dark
green, semi-solid mass (400 gm). The extract so obtained
was chromatographed over silica gel column ( of height
1.2 m with 5 cm diameter) packed with 900g silica gel.
The column was eluted with different solvents in order of
increasing polarity and the following compounds were
isolated, purified and characterized.
i) 5,4´-Dihydroxy-3,6,7,3´,5´ pentamethoxy flavone (III)
This compound was isolated on elution of the column
with chloroform and ethyl acetate in 3:1 ratio. After
removal of solvent, a yellow crystalline solid was
obtained. A single spot was seen on TLC examination.
Its Rf value was found to be 0.32 in system [C6H6:
CH2Cl2: Et2O (1:1:1)]. The melting point of this
compound was 1790 C. IR (KBr, cm–1): 3510 (-OH),
1660, 1600 (C=C-C=O). 1H NMR (δ ppm, CDCl3): 12.32
s (-OH), 7.45 s (H-2´, H-6´), 6.43 s (H-8), 4.01 s (-
OCH3), 3.96 s (-OCH3), 3.95s (2 × OCH3), 3.90s (-
OCH3). 13C NMR (δ ppm, CDCl3): 189 (C=O), 51.6
(3,6,7,3’,5’-OCH3 ),106(C-2),162(C-3), 122 (C-1’),
127(C-2’, C-6’), 159 (C-3’ ,C-5’),162(C-4’), 104 (C-10),
110 (C-6), 132 (C-7), 128(C-5),194(C-8) and 150 (C-9).
MS (m/z): 404 [M+], 389 [M-Me]+, 373 [M-OMe]+, 359,
343, 331, 246, 118, 69, 57 etc.( for molecular formula
C20H20O9).
ii) Isolation of compounds (IV) and (V)
On elution of the column with pure solvent ethyl acetate,
two compounds were obtained which were further
separated by preparative TLC using Petrol-Et2O (7:3)
mixture as solvent system.
Benzyl-2,6-dimethoxy benzoate (IV)
This compound was isolated as colourless oil, 45mg and
it showed a single spot on TLC plate. IR (KBr, cm–1):
3000-2880 (C-H str.), 1750 (COOR), 1610, 1590, 1480,
1400, 1310, 1265 (C-O str.), 1125, 1080 cm–1. 1H NMR
(δ ppm, CDCl3): 7.44 dd (2 × Ar-H), 7.36 m (3 × Ar-H),
7.26 d (1 × Ar-H), 6.55 d (2 × Ar-H), 5.36 s (O-CH2-Ph),
3.80 s (2× OCH3). 13C NMR (δ ppm, CDCl3): 170.92
(C=O), 65.23 (-O-CH2-), 56.3(OCH3 at C-2 and C-6),
103( C-1’) , 135.28 (C-2’,C-6’), 127.34 (C-3’,C-5’),
126.04 (C-4’),162.45 (C-1), 135.69 (C-2 and C-6),130.04
(C-4),102.22 (C-3 and C-5) . MS (m/z): 272 [M+], 181
[M-CH2-Ph]+, 165 [C6H3(OMe)2CO]+, 149 [181-MeOH]+
91[CH2C6H5]+ for molecular formula C16H16O4 .
iii) Benzyl-2,3,5,6-tetramethoxy benzoate (V)
It was isolated as colourless oil, 30 mg and it showed a
homogenous behaviour on TLC plate. IR (KBr, cm–1):
2900-2880 (C-H str.), 1735 (-COOR), 1595, 1380, 1280
(C-O str.), 1130, 1100, 1080 cm–1. 1HNMR (δ ppm,
CDCl3): 7.47 d (2 × Ar-H), 7.35 m (3 × Ar-H), 6.75 s (1 ×
Ar-H), 5.39 s (Ph-CH2-O), 3.85 s (2× OMe), 3.74 s (2 ×
OMe). 13C NMR (δ ppm, CDCl3): 178 (C=O), 67 (-O-
CH2-), 56.4 (OCH3 at C-2, C-3, C-5, C-6), 104( C-1’) ,
136.38 (C-2’,C-6’), 126.04 (C-3’,C-5’), 128.34 (C-
4’),168.30 (C-1), 145.02 (C-2 and C-6),165.32 (C-
4),130.04 (C-3 and C-5). MS (m/z): 332 [M+], 317 [M-
Me]+, 289 [317-CO]+, 225 [C6H(OMe)4CO]+,197 [225-
CO]+,91 [-CH2C6H5]+ for molecular formula C18H20O6.
RESULTS AND DISCUSSION
Compound (III): For this a highly abundant molecular ion
peak was obtained at m/z 404 in the mass spectrum. A
base peak at m/z 389 resulted due to the loss of methyl

32
radical from molecular ion. Other prominent peaks were
373 [M-OMe]+, 359, 343, 331, 246, 118, 69, 57. From
this spectral data its molecular formula was found to be
C20H20O9. It’s IR Spectrum (KBr, cm–1) revealed the
presence of hydroxyl group (3510 cm–1) and conjugated
carbonyl moiety (1660, 1600 cm–1). The 1H NMR
spectrum (δ ppm, CDCl3) confirmed the presence of
chelated hydroxyl group (C-5) by the appearance of the
diagnostic signal at δ 12.32. A sharp singlet integrated for
two protons (H-2´ and H-6´) of ring C and a singlet due
to one proton (H-8) of ring A appeared in the aromatic
region at δ 7.45 and δ 6.43 respectively. Large singlets at
δ 3.90, δ 3.95, δ 3.96 and δ 4.01 corresponded to the five
methoxy groups in the molecule. The 13C NMR spectrum
(δ ppm,CDCl3) corroborated the presence of 20 carbons
ranging from δ 46 to δ 194. The value at δ 189 was
assigned to the carbonyl (C=O) moiety at C-4. The
presence of alkene (C=C) carbons at C-2 and C-3 was
ascertained by signals at δ 106 and δ 162.The absorption
signal for carbon of the methoxy groups appeared at δ
51.6. The carbons of ring B appeared at δ 122 (C-1’),
127(C-2’, C-6’), 159 (C-3’, C-5’) and 162 (C-4’).
Similarly, the ring A carbons showed signals at δ 104 (C-
10), 110 (C-6), 132 (C-7), 128 (C-5), 194(C-8) and 150
(C-9). With the help of above spectral data, compound
(III) was identified as 5,4´-dihydroxy-3,6,7,3´,5´
pentamethoxy flavone. The spectral data was compared
with the literature values. This is the first report of the
presence of this compound in this lichen.
O
H3CO
H3CO
OOCH3
OCH3
OCH3
10
5
6
7
89
1
2
3
1'
6' 5'
4'
3'2'
4
5-Hydroxy-3,6,7,3',4'-pentamethoxy flanone
AB
C
Fig.1. Compound III
Compound (IV) : The mass spectrum of this compound
exhibited a significant molecular ion peak at m/z 272.
Other prominent fragment ion peaks appeared at m/z 181
[M-CH2Ph]+, 165 [C6H3(OMe)2CO]+ (base peak) and
91[C6H5-CH2]+ etc. From this spectral data its molecular
formula was confirmed as C16H16O4. . In the IR spectrum
(KBr, cm–1) the absorption bands at 1750, 1610, 1590 cm–
1 revealed the presence of an aromatic ester function.
Other significant absorption bands appeared at 3000-2880
(for C-H stretching), 1265 (for C-O stretching). The 1H
NMR spectrum (δ ppm,CDCl3) displayed the presence of
a double doublet at δ 7.44 for the two ortho aromatic
protons and a multiplet at δ 7.36 corresponded to the three
aromatic protons of the monosubstituted benzene ring.
The signal for the three aromatic protons of the
trisubstituted benzene ring appeared as a doublet at δ 7.26
for one proton and another doublet at δ 6.55 for
remaining two protons. The benzylic proton gave a
singlet at δ 5.37. For the two methoxy groups a singlet
integrated for six protons appeared at δ 3.8 ppm. In
13CNMR spectrum (δ ppm,CDCl3) the presence of
carbonyl moiety was confirmed by the appearance of a
signal at δ 170.92. Signal for oxymethylene carbon was
obtained at δ 65.23 and the signal for the two methoxy
carbons appeared at δ 56.3. The aromatic carbons of the
benzyl moiety gave absorption signals at δ 103(C-1’),
135.28 (C-2’,C-6’), 127.34 (C-3’,C-5’) and 126.04 (C-
4’). The absorption signals for the aromatic carbons of the
other ring appeared at δ 162.45 (C-1), 135.69 (C-2, C-6),
130.04 (C-4), 102.22 (C-3 and C-5). With the help of
these spectral data this compound was identified as
benzyl-2, 6-dimethoxy benzoate. The spectral data was
compared with reported values. This compound is being
reported for the first time in this lichen.
C
OCH3
OCH3
O
O CH2
32
4
56
11'
2' 3'
4'
5'6'
Benzyl-2,6-dimethoxy benzoate
Fig.2. Compound IV
Compound (V) : The mass spectral study of this
compound revealed a highly abundant molecular ion

33
peak at m/z 332 in addition to other important peaks at
317 [M-Me]+, 289 [317-CO]+ and 91 [C6H5CH2]+ (base
peak) etc. These values suggested its molecular formula
to be C18H20O6. In the IR spectrum (KBr, cm–1) the
absorption bands at 1735 and 1590 cm–1 confirmed the
presence of an aromatic ester function. Other significant
absorption bands were found at 2900-2880 (for C-H
stretching), 1280 (for C-O stretching). The 1H NMR
spectrum (δ ppm, CDCl3) displayed a sharp singlet at δ
6.57 ppm for the lone aromatic proton of the
pentasubstituted benzene ring. A broad doublet at δ 7.47
and a multiplet at δ 7.35 corresponded to the two ortho
and three aromatic protons of the phenyl ring
respectively. A singlet δ 5.39 was assigned to the benzylic
protons. The four methoxy groups appeared at δ 3.74 and
3.85 as singlets, each integrated for 6 protons. The
13CNMR spectrum (δ ppm,CDCl3) showed absorption
signal at δ 178 for carbonyl group ( C=O) and for the
oxymethylene carbon an absorption signal appeared at δ
67.The presence of methoxy carbons was ascertained by
the presence of signal at δ 56.4. Signals for the aromatic
carbons of the benzyl moiety appeared at δ 104 (C-1’),
136.38 (C-2’, C-6’), 126.04 (C-3’,C-5’), 128.34 (C-4’).
The aromatic protons of the other ring were identified by
the presence of signals at δ 168.30 (C-1), 145.02 (C-2, C-
6), 130.04 (C-3,C-5). The absorption signal for C-4
appeared at δ 165.32. From these spectral studies it was
concluded that this compound is benzyl-2,3,5,6-
tetramethoxy benzoate. The spectral data was compared
with literature values. This is the first report of the
presence of this compound in this lichen.
C
O
O CH2
H3CO
H3CO
OCH3
OCH3
32
4
5 6
11'
2' 3'
4'
5'
6'
Benzyl-2,3,5,6-tetramethoxy benzoate
Fig.3. Compound V
The characterization of compound (I) stigma-4,22-diene-
3-one and compound (II) stigmasterol was done on the
basis of IR, 1HNMR, mixed m.p.s. and co-TLC with
authentic samples.
III. ANTIFERTILITY ACTIVITY OF PARMELIA
PERLATA ON MALE ALBINO RATS
Experimental study
Adult proven fertile colony bread Wistar rats weighing
150-175 g, were maintained in poly propyline cages with
rat feed (Hindustan Lever Ltd.) and top water ad libitum.
The animals were divided into one control group and one
experimental group of 10 rats each. The routine dose of
the plant extract was freshly dissolved in 0.5 ml of
distilled water and administrated to test rats- 100 mg/kg
per day. The study was approved by the ethical committee
of the Department of zoology, University of Rajasthan.
Fertility test
The mating tests were performed from days 55 to 60.
Male rats from control and treated groups were caged
overnight with proestrous females in the ratio of 1:2 for
normal mating. Presence of sperm in the vaginal smear
confirmed positive mating and the day was taken as an
index of Day-I of gestation. The implantation sites of
mated females were checked after 2 weeks by
laparotomy. Autopsy were done on mating males.
Blood and serum study
The animals were scarified using light ether anesthesia
after 60 days. Blood was collected through cardiac
puncture and serum was separated. The RBC and WBC
count [14], Haemoglobin [15] and hematocrit [16] were
analysed with routine methods. The serum was also tested
for protein, cholesterol, HDL-cholesterol, phospholipid
and triglyceride.
Sperm analysis
Sperm motility was evaluated in cauda epididymides and
sperm density in cauda epididymides and testes by the
method [17].

34
Tissue Biochemistry and Histology
The testes, epididymides, seminal vesicle and ventral
prostate were dissected out and weighed. Fresh tissue
from testis and accessory sex organs were processed for
the biochemistry estimation of glycogen, protein, sialic
acid, fructose and cholesterol. Beside they were fixed in
Bouins’s fluid, embedded in paraffin, sectioned at 5-6 µm
and stained with hematoxylin and eosin for histological
examination.
Quantitative study
Various testicular cell components i.e. spermatogonia,
sertoli cells, preleptotene, pachytene, secondary
spermatocyte and rounded spermatids were quantitatively
analyzed using 800x. Interstitial cell types such as
mature, fibroblasts, degenerating Leydig cells, were
calculated by applying a differential cell count, which
were statistically verified by the binomial distribution
[18]. The Leydig cell nuclear area and sertoli cells nuclear
area were also measured at 800x and 80x, respectively.
Serum testosterone levels were assessed from samples
using radioimmuno assay method [19].
Statistical analysis
The data were expressed as mean ±standard error of mean
(SEM) and student’s ‘t’ test was used to assess the
statistical significance. The statistical significance level
was set at p<0.01 and p<0.001.
RESULTS
Reproductive organ weights
A significant (p<0.01) reduction in the weight of testes,
epididymides, seminal vesicles, and ventral prostate was
observed in the ether extract of Parmelia perlata (Table-
I).
Sperm Dynamics and fertility
The number of spermatogonia, spermatocyte and
spermatids were significantly reduced (p<0.01) with the
Parmelia perlata extract. The preleptotene, secondary
spermatocyte and step-19 spermatids were decreased by
59.07%, 65.36%, 52.45% respectively and the mature
Leydig cells were also reduced by 41.57%. At this dose
level, Leydig cell nuclear area and cytoplasmic area, as
well as the cross sectional surface area of sertoli cells,
were significantly reduced (p<0.01) when compared to
controls. The sperm density was significantly reduced in
the testes (p<0.01) (Table-II) and (Table-IV).
Biochemical changes
The cholesterol content of the testes was significantly
(p<0.01) increased while the protein and fructose content
was significantly reduced (p<0.01) after following
treatment with Parmelia perlata extract (Table-III).
Blood and serum biochemistry
The RBC and WBC counts, haemoglobin, haematocrit,
blood sugar and the serum protein, cholesterol,
triglyceride, phospholipid and HDL-cholesterol levels
were within the normal range. Serum testosterone level
was decreased.
DISCUSSION
Oral administration of Parmelia perlata reduces the
weight of testis, and secondary sex organ [20]. Marked
decrease in the sperm cell counts specially the number of
secondary spermatocyte and rounded spermatid also
reduces the testis weight [21]. Reduced protein content
may be another reason for low sperm density as the
growth rate of any organ is proportional to its protein
content [22]. Reduction in Leydig cells nuclei diameters
and disintegration of Leydig cells always lead to decrease
in the androgen level [23]. Reduced nuclear area also
manifest the impairment of Leydig cells function and
decrease the number of mature Leydig cells.
Deformations of Leydig cells indicate the insufficiency of
those cells to synthesize testosterone [24-25].
The reduction in sperm density and motility in
cauda epididymides is an important predictor of sperm
fertilizing ability [26] and also suggest low level of
testosterone to epididymis and therefore affect epididymal
function [27]. The impaired epididymal function may also
be due to reduced activity of the testes, which affects the
normal passage to testicular’s fluid into epididymis, this
is also confirmed by decreased epididymal weight.

35
Results also show low counts of sertoli cells and some
structural changes in sertoli cells after administration of
Parmetia perlata extract. The reduction in number of
secondary spermatocyte and spermatids reflected non-
availability of androgen binding protein (ABP) from
Sertoli cells [28]. ABP is essential to maintain intra
testicular androgen concentration and transformation of
advance stages of sperm cells. Meiotic and post meiotic
sperm cells were highly sensitive to androgen
concentration [29] and the alternation of androgen level in
testis may affect the transformation of spermatocyte to
spermatids.
The decreased level of sialic acid content might
alter the structural integrity of acrosomal membrane,
which ultimately affects the metabolism, motility and
fertilizing capacity of spermatozoa [30]. Parmelia extract
administration caused a decreased level in fructose
concentration, which shows the lack of energy source in
female body during capacitation.
In conclusion, the present study shows that the 100
mg/kg /day for 60 days of Parmelia perlata petroleum
ether extract of male albino rats affects their reproductive
efficiency by alteration in the spermatogenic activity
without adverse toxicity.
IV. CONCLUSION
Lichens have tremendous medicinal values which are due
to the presence of various phytochemical constituents.
With the help of chromatographic techniques, we isolated
three compounds (III- V) for the first time from this
lichen. The medicinal values imparted to the lichen by
these compounds can be studied by future researchers. To
investigate the biological activity of this lichen, the anti-
fertility activity of the pet.ether extract was examined
which revealed the anti-spermatogenic activity of the
extract without adverse toxicity. The study suggests
further scope for research on different biological activities
of Parmelia perlata.
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37
Table I
Effect of Parmelia perlata extract on testicular cell population in rats
Testicular cell counts (number/10 cross section)
Sertoli
cell
Spermatogonia
Preleptotene
Pachytene
Secondary
Spermatocytes
Step-19
Spermatids
Control
(distilled water,
0.5 ml)
Parmelia perlata
200 mg/day per
rat
2.81
± 0.02
1.31**
± 0.0828
(-53.3)
6.87
± 0.02
3.037**
± 0.2088
(-55.8)
19.95
± 1.9
7.597**
± 1.169
(-61.9)
29.29
± 0.73
11.669**
± 0.6073
(-60.19)
48.1
± 0.6
20.630**
± 3.640
(-57.12)
34.75
± 0.8
7.076**
± 3.02
(-79.6)
All values are Mean ± S.E.M. (n=10); ns: non significant; Level of significance ** P<0.001 vs control Percent variations Vs control in
parentheses
Table II
Effect of Parmelia perlata extract on body weight, organ weights together with seminiferous tubule and Leydig cell
nuclear diameter
Body wt.
(g)
Organ Weight (mg/100g body weight)
Seminiferous
tubule diameter
(m)
Leydig cell nuclear
area (m)
Control (distilled
water 0.5 ml)
240
± 1.4
1345
± 4.7
529.5
± 1.2
605.7
± 1.2
308.5
± 2.02
268 ± 9
11.1 ± 0.02
Parmelia perlata 200
mg/day per rat
230ns
± 5.8
975**
± 17
404.4
± 9
156.94**
± 31
115.97**
± 5.1
200.4**
± 10.4
5.32 ± 16**
All values are Mean ± S.E.M (n=10); ns: non significant; Level of significance** P<0.001 vs control

38
Table III
Effect of Parmelia perlata extract on tissue biochemistry in rats
Protein (mg/g)
Sialic acid (mg/g)
Glycogen
(mg/g)
Cholestrol
(mg/g)
Fructose
(mg/g)
Testes
Cauda
epididymides
Seminal
vesicles
Ventral
prostate
Testes
Cauda
epididymides
Seminal
vesicle
Ventral
prostat
e
Testes
Testes
Seminal
vesicles
mg/g
Control
Parmelia
perlata
178.8
± 0.5
132.56**
± 1.48
268.7
± 3.6
204.41**
± 1.73
186
± 3.4
134.04**
± 5.9
162.4
± 0.18
141.45**
± 2.96
4.64
± 0.16
3.88*
± 0.14
5.34
± 0.15
3.66**
± 0.06
4.2
± 0.09
3.80ns
± 0.16
5.2
± 0.23
3.72**
± 0.12
4.85
± 0.6
2.01
± 0.01
7.82
± 0.11
15.15**
± 0.6
4.24
± 0.04
3.51**
0.073
All values are expressed as mean ± S.E.M. (n=10); ns=non significant; Level of significance * P<0.01; ** P<0.001 vs control.
Table IV
Effect of Parmelia perlata extract on sperm motility, concentration and fertility in rats
Sperm motility (%)
cauda epididymides
Sperm density (million/ml)
Fertility
Testes
Cauda epididymides
Control
Parmelia perlata
74.1 ± 2.38
29.85 ± 1.15**
4.15 ± 0.21
2.25 ± 0.1**
52.2 ± 2.66
11.8 ± 1.28**
100%(+ve)
100%(+ve)
All values are mean ± S.E.M.; levels of significance ** P<0.001