[P-123]
EFFECTS OF Salvia officinalis L. EXTRACT ON UTERUS OF
OVARIECTOMIZED RAT

Vlaicu ªANDOR, Gabriela BAICEANU, Mircea TAMAª and Tibor L. KRAUSZ
University of Medicine and Pharmacy "Iuliu Hatieganu", 13 Emil Isac street, 3400 Cluj-Napoca, Romania

ABSTRACT

Interactions between estrogens and Salvia officinalis L. on an animal experimental model using ovariectomized rats are obvious on the uterus. Estradiol (E) administered for 3 days, 0.75 micromoles/kg/day elevates significantly all morphometric indexes. Salvia extract alone has no effect on the uterus, but potentiates in a dose-dependent manner the trophic effect of the E. On the other organs both agents has minimal effects.

Keywords: estradiol, Salvia officinalis L. extract, morphometric index, uterus.


INTRODUCTION

Clinical observation concerning neurovegetative changes in climax suggests possible favourable effects of Salvia officinalis L. (SE) to treat endocrine disturbances during this critical period (6). The effects of SE on the uterus and its interaction with the estrogens were studied on an experimental model using ovariectomized rats (1,4).


MATERIAL AND METHOD

We used virgin female Wistar-Bratislava rats weighting 58-106 g (Table 1). Animals were kept in standard laboratory conditions, natural light-dark cycle, and divided in 7 groups. Ovariectomy was performed in the same day, using pentobarbital Na i.p. general anaesthesia (40 mg/kg/2ml). The first group (10 rats) was sham-operated, the following 6 groups (8 rats/group) undergone lumbar ovariectomy after local antiseptic treatment (1). No general antibiotic was used prior or after ovariectomy. After 3 weeks specific treatment started (Table 2).


Table 1. Body-weight
Body-weight g (x ± e.s)
I
II
III
IV
V
VI
VII
79.5
±
3.236
81.375
±
4.288
75.125
±
4.864
71.25
±
3.74
74.75
±
3.11
79.375
±
3.722
78.0
±
5.254
Group I. sham-operated
Group II. ovariectomy (control)
Group III. estradiol (E) i.p. 0.75 micromoles/kg/2ml/day, 3 days
Group IV. (E) (like group III.) and diluted (1:50) Salvia officinalis L. extract (DS) p.o. 1.0 ml/0.1 kg/12 hours, 4 days
Group V. E (like group III and IV) and concentrated (1:5) Salvia officinalis L. extract (CS), p.o. same volume and schedule like group IV.
Group VI. only DS, like group IV.
Group VII. only CS, like group V.

Table 2. Experiment schedule
Time 
Group
(-)3
weeks
0
12h
24h
36h
48h
60h
72h
84h
96h
I (SH)
SH
SS
SS
SS+ES
SS
SS+ES
SS
SS+ES
SS
S
A
C
R
I
F
I
C
E
II (O)
O
SS
SS
SS+ES
SS
SS+ES
SS
SS+ES
SS
III (O+E)
O
SS
SS
SS+E
SS
SS+E
SS
SS+E
SS
IV (O+E+DS)
O
DS
DS
DS+E
DS
DS+E
DS
DS+E
DS
V (O+E+CS)
O
CS
CS
CS+E
CS
CS+E
CS
CS+E
CS
VI (O+DS)
O
DS
DS
DS+ES
DS
DS+ES
DS
DS+ES
DS
VII(O+CS)
O
CS
CS
CS+ES
CS
CS+ES
CS
CS+ES
CS
Legend:
SH - sham-operated  E - Estradiol; 0.75 micromoles/kg/2 ml/day, 3 days
O - ovariectomized  DS - diluted Salvia extract (1:50) p.o. 1 ml/kg/12 h
SS - Salvia solvent  CS - concentrated Salvia extract (1:5) p.o. 1.0 ml/kg/12 h
S - Estradiol solvent

Animals were sacrificed at 12 hours after the last dose using pentobarbital Na i.p. 70 mg/kg/2ml. All organs taken into study were measured on a torsion balance, than dried (at 110°C) and measured again, so we obtained 3 weights: total, dry and water-content (by difference). Relative weight was calculated for each organ (organ weight in mg/100 g body-weight)(as shown in Table 3). Student's "t" test was used to test for statistical significance (p<0.05).

Medication used

  1. Pentobarbital Na aqueous solution 2%, prepared extemporaneously
  2. Estradiol (E): ampoules(Biofarm®, Romania) of 2,5 mg/1ml, diluted in his own excipient (hexilic alcohol 5% in oleum helianthii)
  3. Salvia extract (SE), obtained from the Pharmaceutical Botany Dept. of the "Iuliu Hatieganu" UMF; two dilutions were prepared - 1:5 and 1:50
RESULTS AND DISCUSSION

In our experiment, estradiol elevates all 3 morphometric indexes, and causes liquid accumulation in the uterine horn. Salvia extract (SE) alone does not have any effect on the uterus, but it is able to potentiate estradiol action (Tables 3 and 4).


Table 3. Organ relative weight
 
Relative weight mg/100g (x±e.s)
1
2
3
4
5
6
7
Uterus
Total
50.968
±
4.273
46.925
±
3.092
137.258
±
9.126
172.678
±
6.564
183.825
±
17.448
57.054
±
4.638
54.351
±
5.663
D
43.373
±
4.681
40.003
±
2.634
116.246
±
7.712
148.711
±
8.202
153.441
±
16.183
48.405
±
3.442
44.280
±
3.66
Dry
7.595
±
1.212
6.922
±
1.247
21.012
±
2.451
23.967
±
3.205
30.384
±
3.606
8.649
±
1.438
10.071
±
4.687
Heart
Total
437.981
±
24.409
444.366
±
15.216
422.994
±
15.237
495.546
±
7.779
476.615
±
10.941
456.977
±
17.106
444.06
±
19.692
D
354.705
±
22.457
359.553
±
12.7
343.114
±
12.578
408.264
±
7.826
390.426
±
9.764
377.759
±
14.782
361.372
±
16.97
Dry
83.276
±
2.235
84.813
±
2.85
79.88
±
3.207
87.281
±
1.752
86.189
±
1.664
79.218
±
2.586
82.688
±
3.008
Submax-illary glands
Total
226.263
±
10.34
214.993
±
11.491
213.599
±
15.802
229.098
±
8.174
222.94
±
8.173
216.987
±
5.467
224.681
±
7.407
D
182.371
±
7.982
170.872
±
9.486
172.201
±
12.009
182.363
±
6.234
173.033
±
5.526
169.457
±
3.94
183.675
±
5.693
Dry
43.892
±
2.92
44.121
±
2.182
41.398
±
4.136
46.735
±
2.016
49.907
±
3.127
47.53
±
1.692
41.006
±
2.132
Adrenals
Total
24.505
±
2.088
21.336
±
2.5
27.193
±
1.717
25.73
±
2.648
33.48
±
1.506
28.045
±
1.252
27.417
±
1.606
D
16.993
±
1.686
15.24
±
2.281
19.582
±
1.2
17.659
±
1.755
24.215
±
1.128
19.421
±
1.162
19.194
±
1.293
Dry
75.148
±
0.532
6.096
±
0.577
7.611
±
0.584
8.071
±
0.938
9.265
±
0.867
8.624
±
0.192
8.293
±
0.493
Liver
Total
4785.028
±
280.536
4904.815
±
179.846
5043.896
±
267.192
5904.824
±
693.525
6021.0
±
225.9
4740.996
±
380.475
5904.47
±
430.477
D
3910.268
±
254.189
3997.741
±
163.802
4030.144
±
251.206
4871.716
±
673.44
4983.595
±
190.667
3778.6
±
312.725
4794.619
±
375.435
Dry
874.76
±
51.009
907.074
±
54.345
1013.752
±
81.485
1030.108
±
33.724
1037.405
±
60.569
962.396
±
71.165
1109.851
±
60.407

Table 3. Organ relative weight (continued)
 
Relative weight mg/100g (x±e.s)
1
2
3
4
5
6
7
Kidneys
Total
955.332
±
31.599
942.007
±
32.603
1069.652
±
43.917
1075.082
±
42.5
1097.047
±
49.232
906.753
±
34.287
1014.804
±
53.986
D
749.353
±
25.621
734.919
±
25.321
840.741
±
34.058
839.977
±
33.557
856.152
±
38.158
704.409
±
27.295
791.007
±
43.281
Dry
205.979
±
6.182
207.088
±
7.474
228.911
±
10.285
235.105
±
9.38
240.894
±
11.143
202.344
±
6.674
223.798
±
10.876
Stomach
Total
1026.348
±
38.612
952.138
±
71.899
971.533
±
36.005
1116.501
±
38.232
1027.279
±
45.819
1013.755
±
62.124
1060.029
±
54.976
D
837.696
±
38.682
797.917
±
69.163
795.737
±
28.99
841.75
±
76.776
847.372
±
41.927
837.632
±
48.702
887.967
±
47.254
Dry
188.652
±
8.561
154.221
±
6.095
175.796
±
7.356
274.751
±
76.207
179.907
±
5.947
176.123
±
14.035
172.062
±
8.918
Spleen
Total
567.898
±
17.64
695.083
±
41.056
508.331
±
34.616
623.955
±
46.515
644.022
±
57.226
538.127
±
31.052
791.145
±
122.936
D
444.154
±
13.915
544.45
±
33.066
398.201
±
26.832
483.008
±
36.058
498.837
±
43.674
416.908
±
23.217
619.967
±
96.97
Dry
123.744
±
3.759
150.633
±
8.06
110.13
±
7.92
140.947
±
10.54
145.185
±
13.574
121.219
±
7.898
171.178
±
25.978

Literature regarding estrogenic action of Salvia officinalis L. is quite vague (2). Some improvements were observed in climax after such treatment (6). Effects of SE depend on its composition. The plant contains a high diversity of compounds (5,7). Some are toxic in high dosage, like essential oils. Sabinyl acetate, present in essential oils of other Salvia species (S. lavandulifolia), can be abortifacient in female rat (3). Our hydro-alcoholic extract of Salvia officinalis L. was obtained by percolation and does not contain essential oil (5).

A pertinent analysis regarding the uterine effects of SE requires the exact composition of the Salvia extract, eventually individual testing of each component using this experimental model.

On the other organs estradiol and SE have little influence (Tables 3 and 4).


CONCLUSIONS
  1. Estradiol and Salvia officinalis L. extract administered to virgin female ovariectomized rats has distinct effects on uterus and the rest of the organs taken into study

  2. Estradiol at a dose of 0.75 micromoles/kg/day, for 3 days elevates significantly the relative weights (total, dry and water-content).

  3. Salvia extract in two concentrations does not alter uterine relative weight.

  4. Associated to estradiol, Salvia extract potentiates in a dose-dependent manner its trophic effect.

  5. On the other organs estradiol has slight and frequently discrepant effect on the 3 types of weight.

  6. Salvia extract in these two concentrations has minimal effect on the morphometric parameters on these organs.

Table 4. Statistical significance
"t" Student
        p
Relative weight
Total
D
Dry
UTERUS II-III
<0.001
<0.001
<0.001
II-IV
<0.001
<0.01
<0.001
II-V
<0.001
<0.002
<0.01
III-V
<0.05
n.s.
n.s.
IV-VI
<0.001
<0.01
<0.001
V-VII
<0.001
<0.02
<0.001
HEART II-IV
<0.05
n.s.
<0.02
II-V
<0.05
n.s.
<0.05
III-IV
<0.002
n.s.
<0.002
IV-VI
<0.02
<0.01
<0.05
SUBMAX. V-VII
n.s.
<0.01
n.s.
ADRENALS II-III
n.s.
<0.01
n.s.
II-V
<0.01
<0.05
<0.01
II-VI
n.s.
<0.01
n.s.
II-VII
n.s.
<0.05
n.s.
III-V
<0.05
n.s.
<0.02
V-VII
<0.01
n.s.
<0.001
LIVER II-V
<0.01
n.s.
<0.01
III-V
<0.002
n.s.
<0.01
KIDNEY II-IV
<0.02
<0.05
<0.02
II-V
<0.05
<0.05
<0.02
IV-VI
<0.05
<0.05
<0.05
STOMACH I-II
n.s.
<0.01
n.s.
II-III
n.s.
<0.02
n.s.
II-V
n.s.
<0.01
n.s.
III-IV
<0.01
n.s.
n.s.
SPLEEN II-III
<0.02
<0.02
<0.02
II-VI
<0.05
<0.05
<0.05
III-IV
n.s.
<0.05
n.s.
n.s.- not significant

REFERENCES
  1. Baiceanu G.(1999) Efecte endocrine ale Salvia officinalis L. MD thesis, UMF "Iuliu Hatieganu" Cluj-Napoca.

  2. Benigni R., Capra C., Cattorini P.E.(1962), Piante medicinali, vol. 2, Inverni &Della Beffa, Milano, 1409-1420.

  3. Fournier G., Pages N., Cosperec I. (1993): Contribution to the study of Salvia lavandulifolia essential oil: Potential toxicity attributable to sabinyl acetate. Planta Med.59, 96-97.

  4. May M. (1971): Estrogenic and antiestrogenic agents, In Turne R.A., Hebborn P. (Eds.), Screening methods in Pharmacology vol.2, New York-London, 85-100.

  5. Tamas M., Fagarasanu E., Ionescu C. (1986): Contributii la studiul fitochimical produsului Salviae folium, Farmacia 34,181-186.

  6. Vinti D., Raican D., Tamas N., Toader S. (1985): Aprecierea efectului extractului de Salvia in tratamentul unor tulburari de climacteriu. Simpozion. Medicamente indigene de origine vegetala UMF Cluj-Napoca Abstract Book p. 59.

  7. Wang M F., Kikuzaki H., Zhu N.Q., Sang S.M., Nakatani N., Ho C.T. (2000): Isolation and structural elucidation of two new glycosides from sage (Salvia officinalis L.) J.Agr.Food Chem. 48, 235-238.

[P-123]