Journal List > J Nutr Health > v.50(3) > 1081498

Ko, Liu, and Kim: Effect of oral guava leaf extract administration on antioxidant and vasculoprotective activity in ovariectomized rats

Abstract

Purpose

The aim of this study was to assess the effects of guava leaf extract (GLE) supplementation on antioxidant enzyme activity and expression of endothelial nitric oxide synthase (eNOS) mRNA in ovariectomized rats. Methods: All animals were randomly assigned to four groups (n = 7 for each group): non-ovariectomized control (Sham), the ovariectomized control (OVX), ovariectomy + 150 mg/kg b.w. of GLE (OVX·GL), and ovariectomy + 300 mg/kg b.w. of GLE (OVX·GH). Treatment groups were administered GLE for 8 weeks every day. Results: Body weight gain was significantly reduced in the OVX·GL group compared with the OVX group (p < 0.05). The level of serum 17β-estradiol (E2) was significantly lower in the OVX groups than the Sham group (p < 0.05). Serum triglyceride (TG) and HDL-cholesterol levels were not significantly different between all groups. However, serum total cholesterol (TC) level was significantly reduced in the OVX·GH group compared with the OVX group (p < 0.05). Serum free fatty acid (FFA) level and liver TG level were significantly reduced in both OVX·GL and OVX·GH groups compared with the OVX group (p < 0.05). Furthermore, serum glutathione peroxidase (GPx) activity was significantly elevated in the GLE groups (p < 0.05). The mRNA expression level of GPx was not affected by ovariectomy. However, administration of GLE resulted in significantly increased nuclear factor erythroid 2-related factor (Nrf2) and catalase (CAT) mRNA expression levels in the liver (p < 0.05). In addition, liver nitric oxide (NO) level was significantly reduced in the OVX·GH group compared with the OVX group (p < 0.05). Expression level of endothelial nitric oxide synthase (eNOS) was significantly elevated in the OVX·GH group compared with the OVX group (p < 0.05). Conclusion: These results suggest that GLE could have protective effects in OVX rats by stimulating eNOS expression and improving the antioxidant defense system.

References

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Fig. 1.
Liver malondialdehyde and nitric oxide levels of each group. (a) Liver malondialdehyde, (b) Liver nitric oxide. Sham; non-ovariectomized control, OVX; ovariectomized control, OVX· GL; ovariectomy + 150 mg/b.w. guava leaf extract, OVX·GH; ovariectomy + 300 mg/b.w. guava leaf extract, ns; not significant. Values are mean ± SD (n = 7 for each group). The different letters (a > b > c) within a column indicate significant differences (p < 0.05) determined by Duncan's multiple range.
jnh-50-236f1.tif
Fig. 2.
Serum catalase and glutathione peroxidase levels of each group. (a) Serum catalase, (b) Serum glutathione peroxidase. Sham; non-ovariectomized control, OVX; ovariectomized control, OVX· GL; ovariectomy + 150 mg/b.w. guava leaf extract, OVX·GH; ovariectomy + 300 mg/b.w. guava leaf extract, ns; not significant. Values are mean ± SD (n = 7 for each group). The different letters (a > b > c) within a column indicate significant differences (p < 0.05) determined by Duncan's multiple range test.
jnh-50-236f2.tif
Fig. 3.
mRNA expression of Nrf2, CAT, GPx and eNOS in liver of each group. (a) Nrf2; nuclear factor erythroid 2-related factor, (b) CAT; catalase, (c) GPx; glutathione peroxidase, (d) eNOS; endothelial nitric oxide synthase. 1) Sham; non-ovariectomized control, OVX; ovariectomized control, OVX· GL; ovariectomy + 150 mg/b.w. guava leaf extract, OVX·GH; ovariectomy + 300 mg/b.w. guava leaf extract, 2) ns; not significant. Values are mean ± SD (n = 7 for each group). The different letters (a > b > c) within a column indicate significant differences (p < 0.05) determined by Duncan's multiple range test.
jnh-50-236f3.tif
Table 1.
Sequences of primers for RT-PCR
Gene Primer Molecule sequence (5'-3') Annealing Cycle
Nrf21) Forward CCATTTACGGAGACCCAC 55oC 30
Reverse TGAGCGGCAACTTTATTC
GPx2) Forward CTCTCCGCGGTGGCACAGT 58oC 30
Reverse CCACCACCGGGTCGGACATAC
CAT3) Forward GCGAATGGAGAGGCAGTGTAC 55.5oC 30
Reverse GAGTGACGTTGTCTTCATTAGCACTG
eNOS4) Forward ATGGATGAGCCAACTCAAGG 59oC 45
Reverse CTGTCCTCAGGAGGTCTTGC
GAPDH5) Forward CCTCTCTCTTGCTCTCAGTAT 56oC 33
Reverse GTATCCGTTGTGGATCTGACA

1) nuclear factor erythroid 2–related factor

2) Glutathione peroxidase

3) Catalase

4) Endothelial nitric oxide synthase

5) Glyceraldehyde-3-phosphate dehydrogenase

Table 2.
Serum estradiol, body weight, food intake, food efficiency ratio, and organ weight1)
  Sham OVX OVX·GL OVX·GH
Estradiol 2.31 ± 0.28a4) 1.92 ± 0.24b 1.81 ± 0.28b 1.42 ± 0.27c
 Body weight (g)        
 Initial 210.49 ± 5.79ns2) 212.20 ± 7.12 218.70 ± 9.72 200.36 ± 16.33
 Final 264.87 ± 9.95c 359.63 ± 19.57a 302.47 ± 24.17b 323.10 ± 42.61b
 Body weight gain 54.38 ± 6.30b 147.43 ± 18.29a 83.76 ± 30.36b 122.74 ± 49.48a
Food intake (g/day) 14.88 ± 4.34ns 13.62 ± 0.38 13.33 ± 0.97 13.13 ± 0.93
FER3) 0.08 ± 0.02a 0.23 ± 0.03b 0.14 ± 0.05a 0.20 ± 0.08ab
Organ weight (g)        
 Liver 7.21 ± 0.60b 8.80 ± 0.95a 7.73 ± 0.46b 7.62 ± 0.95b
 Uterus 0.54 ± 0.09a 0.13 ± 0.05c 0.37 ± 0.18ab 0.25 ± 0.27bc
 Uterin fat 3.40 ± 1.16b 7.37 ± 2.26a 6.04 ± 1.07a 6.67 ± 2.74a

1) Sham; non-ovariectomized control, OVX; ovariectomized control, OVX·GL; ovariectomy + 150 mg/b.w. guava leaf extract, OVX·GH; ovariectomy + 300 mg/b.w. guava leaf extract

2) ns; not significant

3) FER; food efficiency ratio

4) Values are mean ± SD (n = 7 for each group). The different letters (a > b > c) within a column indicate significant differences (p < 0.05) determined by Duncan's multiple range test.

Table 3.
Serum lipid profile and liver triglyceride level1)
  Sham OVX OVX·GL OVX·GH
Serum (mg/dl)        
 Triglyceride 130.96 ± 13.22ns2) 160.61 ± 71.21 159.47 ± 14.03 153.74 ± 9.37
 TC3) 175.61 ± 14.45b8) 207.28 ± 29.64a 195.99 ± 11.99ab 181.48 ± 22.22b
 HDL-C4) 73.71 ± 5.59ns 75.22 ± 11.75 71.60 ± 5.61 67.49 ± 6.35
 LDL-C5) 75.71 ± 9.28b 99.54 ± 23.25a 92.49 ± 10.56ab 83.24 ± 24.29ab
 VLDL-C6) 26.19 ± 2.64ns 32.12 ± 14.24 31.89 ± 2.81 30.75 ± 1.87
 FFA7) 164.60 ± 61.95b 234.36 ± 77.06a 59.36 ± 47.48c 54.12 ± 27.47c
Liver (mg/g)        
 Triglyceride 402.61 ± 140.10ab 532.28 ± 169.23a 267.89 ± 87.27b 309.14 ± 48.84b

1) Sham: non-ovariectomized control, OVX: ovariectomized control, OVX·GL: ovariectomy + 150 mg/b.w. guava leaf extract, OVX·GH; ovariectomy + 300 mg/b.w. guava leaf extract

2) ns; not significant

3) TC; total cholesterol

4) HDL-C; high density lipoprotein cholesterol

5) LDL-C; low density lipoprotein cholesterol

6) VLDL-C; very low density lipoprotein cholesterol

7) FFA; free fatty acid

8) Values are mean ± SD (n = 7 for each group). The different letters (a > b > c) within a column indicate significant differences (p < 0.05) determined by Duncan's multiple range test.

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