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PHYTOTHERAPY RESEARCH

Phytother. Res. 13, 138141 (1999)

Effect of Parsley (Petroselinum crispum) on the


Skin of STZ Induced Diabetic Rats

T. Tunal,1 A. Yarat,1* R. Yanardag,2 F. Ozcelik,1 O. Ozsoy,2 G. Ergenekon3 and N. Emekli1


1
Department of Biochemistry, Faculty of Dentistry, Marmara University, Nisantas 80200 Turkey
2
Department of Chemistry, Faculty of Engineering, Istanbul University, Avclar 34850 Turkey
3
Dermatology Clinic, Florence Nightingale Hospital, Istanbul, Turkey

Parsley (Petroselinum crispum) is one of the medicinal herbs used by diabetics in Turkey and it has been
reported to reduce blood glucose. The purpose of this study therefore was to investigate the effect of
feeding parsley on diabetes induced impairments in rat skins. Uncontrolled induced diabetes caused sig-
nificant increases in nonenzymatic glycosylation of skin proteins, lipid peroxidation and blood glucose.
Administration of parsley extract did not inhibit these effects except for the increase in blood glucose.
SDS-polyacrylamide gel electrophoresis revealed no significant differences in any protein bands between
any of the groups. Copyright # 1999 John Wiley & Sons, Ltd.
Keywords: skin; diabetes; parsley; nonenzymatic glycosylation; lipid peroxidation; protein electrophoresis.

INTRODUCTION hyperlipaemic (Yazcoglu and Tuzlac, 1996), antihepa-


totoxic (Ozturk et al., 1991), diuretic (Baytop 1984;
Marczal et al., 1997) and rheumatoid (Baytop, 1984)
In diabetes increases in nonenzymatic glycosylation effects have been shown. It has also been reported to be
(NEG) of skin proteins and lipid peroxidation (LPO) used in treatments for lumbago, high blood pressure,
damage skin integrity and cause a decreased defence eczema, kneeache, impotence (Yazcoglu and Tuzlac,
function of the skin against harmful substances (Good- 1996) and nose bleeding (Merzouki et al., 1997).
field and Millard, 1988; Sueki et al., 1989; Bannai et al., Phytochemical screenings of parsley have revealed the
1992; Hasegawa et al., 1992; Chait and Bierman, 1994; presence of flavonoids (Hahlbrock, 1981), coumarins
King and Branskota, 1994; Shindo et al., 1994; Mackool (Anand et al., 1981), apiol, myristicin (Ashraf et al.,
et al., 1994; Maroova et al., 1995; Sakata et al., 1995; 1980; Simon and Quinn, 1988), carotenoids (Francis and
Sibbald and Landolt, 1996). Very good glycaemic control Isaksen, 1989), monoterpenes (a pinene) (Lamarti et al.,
is necessary to prevent these diabetic complications. 1991), sesquiterpenes, phthalides, furanocoumarins
In recent years the consumption of certain herbs as a (Spraul et al., 1991) and ascorbic acid (Davey et al.,
folk remedy for diabetes has increased, and particularly 1996).
because they promise oral treatment, these plants have The present study using STZ induced diabetic rats
stimulated considerable interest (Ivorra et al., 1989; Neff investigated the extent to which feeding parsley extract
et al., 1995; El-Shabrawy and Nada, 1996; Noel et al., inhibited diabetes induced changes in skin NEG, LPO
1997). and electrophoretic patterns of skin proteins.
The following plants, known to lower blood glucose
levels in some diabetic patients, have been investigated in
Turkey: Cerasus mahalep seeds (Akev et al., 1990
1992), Rose hips (Can et al., 1994), Laurus nobilis L. MATERIALS AND METHODS
(Yanardag and Can 1994), Beta vulgaris L. var cicla
(chard) (Yanardag and Colak, 1995), Petroselinum Preparation of aqueous parsley extract. The parsley
crispum (parsley) (Yanardag and Ozsoy, 1995), Zizyphus leaves were collected from Istanbul, Buyukcekmece in
jujuba (Erenmemisoglu et al., 1995), Oleum origani June and July. They were carefully washed with tap water
(Lermioglu et al., 1997) and Rosmarinus officinalis and left to dry in the shade at room temperature. They
(Erenmemisoglu et al., 1997). were then stored in well sealed cellophane bags. 100 g
The hypoglycaemic activity of parsley (Petroselinum quantities of dried parsley leaves were extracted by
crispum) from the family Umbellifera has been shown by adding 1000 mL distilled water and boiling for 30 min.
many researchers (Yanardag and Ozsoy, 1995; Yazco- The extract was then filtered, and the filtrates were
glu and Tuzlac, 1996; Noel et al., 1997). Besides its evaporated under reduced pressure to dryness. The
hypoglycaemic activity, antimicrobial (Manderfeld et al., extract was dissolved in distilled water before adminis-
1997), antianaemic (Baytop, 1984), anticoagulant, anti- tration to normal and STZ induced rats.

* Correspondence to: Dr Aysen Yarat, Marmara Universitesi Dishekimligi


Animals and treatment. Sixty-two male, 6-month-old,
Fakultesi Biyokimya Birimi Guzelbahce, Buyukciftlik sok. No. 6 Nisantasi Swiss albino rats were used in this study. They were
(80200), Istanbul, Turkey. divided into four groups designated as follows: C,

CCC 0951418X/99/02013804 $17.50


Copyright # 1999 John Wiley & Sons, Ltd. Accepted 22 July 1998
EFFECT OF PARSLEY ON DIABETIC RAT SKIN 139

Table 1. Mean levels of body weights at 0 and 42 days and blood glucose at 0, 14 and 42 days for all groups. STZ was given to
the diabetic groups on day 0 (C, control; Cp, control parsley; D, diabetic; Dp, diabetic parsley; SD, standard
deviation)
C (n = 13) Cp (n = 16) D (n = 17) Dp (n = 16) p values
Mean SD Mean SD Mean SD Mean SD (ANOVA)
Weight (g)
Day 0 259.63 26.36 259.87 37.13 255.77 30.80 256.83 20.36 0.973
Day 42 259.02 22.53 257.61 35.26 182.91 34.30 189.53 31.19 0.0001
p values (t test) 0.950 0.861 0.0001 0.0001
Blood glucose (mg/dL)
Day 0 71.33 17.97 71.53 24.62 71.05 21.06 70.49 18.99 0.999
Day 42 73.84 11.96 66.66 10.16 150.33 86.44 110.87 45.18 0.0001
p values (t test) 0.678 0.471 0.001 0.003
Day 0 71.33 17.97 71.53 24.62 71.05 21.06 70.49 18.99 0.999
Day 14 77.20 14.14 75.08 17.29 182.66 55.42 167.29 47.58 0.0001
p values (t test) 0.453 0.641 0.0001 0.0001
Day 14 77.20 14.14 75.08 17.29 182.66 55.42 167.29 47.58 0.0001
Day 42 73.84 11.96 66.66 10.16 150.33 86.44 110.87 45.18 0.0001
p values (t test) 0.520 0.104 0.203 0.002

control; Cp, control parsley; D, diabetic; Dp, diabe- determined by using the method of Ledwozyw et al.
tic parsley. The rats in the diabetic groups were fasted (1986). Electrophoretic examination of skin proteins was
for 18 h and rendered diabetic on day 0 by one carried out by Laemmli SDS-polyacrylamide gel electro-
intraperitoneal injection of 65 mg kg1 STZ (streptozo- phoresis (Laemmli, 1970; Yarat et al., 1995).
tocin, Sigma) in a freshly prepared citrate buffer
(pH = 4.5). Fourteen days later, parsley extracts were Statistical analysis. The result were evaluated using an
fed at 2 g kg1 by gavage, daily for 28 days, to the parsley unpaired t-test, ANOVA variance analysis and regression
groups (Cp,Dp). On day 42 after STZ injection, after an analysis using the NCSS statistical computer package.
18 h period of fasting, tail blood samples were taken from
all rats under ether anaesthesia. The rats were then killed
by giving excessive ether, and skin samples (1  1 cm)
were taken from the back of each rat after removing local RESULTS AND DISCUSSION
fur. Fasting blood glucose (after an 18 h period of fasting)
was determined in the blood samples by the o-toluidine The mean levels of blood glucose and body weights for
method (Relander and Raiha, 1963). the four groups are given in Table 1. Prior to inducing
After removing the dermis, the epidermis of each rat diabetes the groups were checked for differences in
skin sample was homogenized in 3 mL of saline solution weight and blood glucose but none were found
and kept in a deep-freeze. Skin total protein levels were (panova = 0.973, 0.999, Table 1). On day 14 immediately
measured by the method of Lowry et al. (1951). Skin before parsley administration the blood glucose levels
protein nonenzymatic glycosylation levels were were determined. The differences between either diabetic
measured by the 2-thiobarbituric acid method (Parker and either control group levels were being significant
et al., 1981; Yarat et al., 1995). Skin LPO levels were (panova = 0.0001, Table 1). On day 42 body weight

Table 2. Mean levels of skin NEG and LPO for all groups (SD, standard deviation, MDA, malondialdehyde)
C (n = 13) Cp (n = 16) D (n = 17) Dp (n = 16) p values
Mean SD Mean SD Mean SD Mean SD (ANOVA)
NEG (nmol fructose/mg protein) 1.72 0.762.08 1.10 7.06 2.62 7.87 1.92 0.0001
0.327 0.321
<> <>
p values (t test)
0.0001
<>
0.0001
<>
LPO (nmol MDA/mg protein) 0.19 0.05 2.00 0.06 0.31 0.20 0.35 0.14 0.002
0.538 0.469
<> <>
p values (t test)
0.047
<>
0.0001
<>

Copyright # 1999 John Wiley & Sons, Ltd. Phytother. Res. 13, 138141 (1999)
140 T. TUNALI ET AL.

the skins and thickening of the fur on all rats in both


diabetic groups. Table 2 shows values and differences
between groups for skin NEG and LPO levels at 42 days.
NEG and LPO levels were significantly higher than
control (C) in both diabetic (D) groups (panova = 0.0001,
0.002 respectively). Our findings were consistent with
others (Sueki et al., 1989; Goodfield and Millard, 1988;
Figure 1. SDS-PAGE patterns for skin proteins (a) and Maroova et al., 1995; Sakata et al., 1995; Hasegawa
standard mixture (b) (Mw: Molecular weight).
et al., 1992; Shindo et al., 1994). There have been
no prior studies related to parsley and the skin. We
found that the administration of parsley extract did not
(BW) was significantly lower (pt test = 0.0001) and blood inhibit these increases in NEG and LPO levels (pt
glucose (BG) significantly higher than at day 0 test = 0.321, 0.469 respectively) and did not affect them
(pt test = 0.001) for the non parsley diabetic group (D). significantly in controls (C,Cp) (pt test = 0.327, 0.538
In the Dp group BW and BG significantly changed over respectively).
the 42 days (pt test = 0.0001, 0.003, Table 1) and the The protein bands obtained by SDS-PAGE were in the
reduction in BG between days 14 and 42 was significant same position for every sample and molecular weights
(pt test = 0.002) showing the parsley to inhibit this adverse ranging from 43 381 kDa. There were, however, no
effect. This is consistent with the study of Yanardag and significant differences in concentration in any of the
Ozsoy (1995). Yanardag and Ozsoy reported that bands between any of the groups (Fig. 1).
2 g kg1 doses of aqueous extracts of dried parsley No significant correlation was found between any of
leaves decreased the blood glucose of alloxan induced the skin parameters except between NEG and LPO
diabetic rabbits. The antidiabetic properties of parsley (r = 0.5574).
may be due to its terpenoid, flavonoid or ascorbic acid In summary, uncontrolled induced diabetes caused a
contents. Parsley may decrease blood sugar by increasing significant increase in skin LPO, NEG of the skin proteins
insulin secretion from b cells of the pancreas, however, and blood glucose and the administration of parsley
its chemical structure is not well known and requires extracts did not inhibit these effects except for the
further investigations. increase in blood glucose.
Macroscopic evaluation revealed yellow porosities on

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Copyright # 1999 John Wiley & Sons, Ltd. Phytother. Res. 13, 138141 (1999)

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