Korean J Physiol Pharmacol.  2018 Jul;22(4):409-417. 10.4196/kjpp.2018.22.4.409.

Supplementing punicalagin reduces oxidative stress markers and restores angiogenic balance in a rat model of pregnancy-induced hypertension

• Wang Y
• Huang M
• Yang X
• Yang Z
• Li L
• Mei J

• KMID: 2414265
• DOI: http://doi.org/10.4196/kjpp.2018.22.4.409

Abstract

Pre-eclampsia (PE) is a pregnancy disorder that is characterised by severe hypertension and increased risks of foetal and maternal mortality. The aetiology of PE not completely understood; however, maternal nutrition and oxidative stress play important roles in the development of hypertension. The treatment options for PE are currently limited to anti-hypertensive drugs. Punicalagin, a polyphenol present in pomegranate juice, has a range of bioactive properties. The effects of supplementation with punicalagin on angiogenesis and oxidative stress in pregnant rats with induced hypertension were investigated. The pregnant rats were randomly divided into five experimental groups (n=12 per group). Hypertension was induced using an oral dose of NG-nitro-L-arginine methyl ester (L-NAME, 50 mg/kg/day) on days 14-19 of pregnancy. Punicalagin (25, 50 or 100 mg/kg) was given orally on days 14-21 of pregnancy. Punicalagin treatment at the tested doses significantly reduced diastolic, systolic, and mean arterial blood pressure in L-NAME treated rats from day 14. Punicalagin also restored angiogenic balance by increasing the expression of vascular endothelial growth factor and downregulating vascular endothelial growth factor receptor-1/fms-like tyrosine kinase-1. Punicalagin, significantly increased the placental nitric oxide levels as compared to PE group. The increased levels of oxidative stress in rats with PE were markedly decreased by treatment with punicalagin. Punicalagin at the tested doses markedly (p < 0.05) enhanced the placental antioxidant capacity in L-NAME-treated rats. The raised catalase activity observed following L-NAME induction was significantly (p < 0.05) and restored to normal activity levels in punicalagin treatment. Further, 100 mg dose of punicalagin exhibited higher protective effects as compared to lower doses of 25 and 50 mg. This study shows that supplementation with punicalagin decreased blood pressure and oxidative stress and restored angiogenic balance in pregnant rats with induced PE.

Keyword

Angiogenic balance; Gestational hypertension; Oxidative stress; Pre-eclampsia; Punicalagin

MeSH Terms

Animals
Antihypertensive Agents
Arterial Pressure
Blood Pressure
Catalase
Female
Hypertension
Hypertension, Pregnancy-Induced*
Maternal Mortality
Models, Animal*
NG-Nitroarginine Methyl Ester
Nitric Oxide
Oxidative Stress*
Pre-Eclampsia
Pregnancy
Punicaceae
Rats*
Tyrosine
Vascular Endothelial Growth Factor A
Antihypertensive Agents
Catalase
NG-Nitroarginine Methyl Ester
Nitric Oxide
Tyrosine
Vascular Endothelial Growth Factor A

Figure

• 

  Fig. 1 Punicalagin decreased systolic (a) and diastolic (b) blood pressure and also decreased mean arterial pressure (c).Values are expressed as means±standard deviation (SD), n=6; one-way analysis of variance (ANOVA) followed by Duncan's multiple range test (DMRT) showed p<0.05; *p<0.05 vs. control; the letters a–e indicate mean values from experimental groups that differ significantly, p<0.05.

• 

  Fig. 2 Punicalagin increased placental nitric oxide (NO) levels.Values are expressed as means±SD, n=6; one-way ANOVA followed by DMRT showed p<0.05; *p<0.05 vs. control; #p<0.05 vs. NG-nitro-L-arginine methyl ester (L-NAME); the letters a–e indicate mean values from experimental groups that differ significantly, p<0.05.

• 

  Fig. 3 Punicalagin enhanced the antioxidant capacity of the placenta (a), Punicalagin decreased the level of thiobarbituric acid reactive substances (TBARS) (b) and also decreased protein carbonyl levels in the placenta (c) and restored normal catalase activity levels (d).Values are expressed as means±SD, n=6; one-way ANOVA followed by DMRT showed p<0.05; *p<0.05 vs. control; #p<0.05 vs. L-NAME; the letters a–e indicate mean values from experimental groups that differ significantly, p<0.05.

• 

  Fig. 4 mRNA levels (a) and protein expression (b) show that punicalagin restored angiogenic balance.Reverse transcription polymerase chain reaction (RT-PCR) analysis revealed a marked decrease in vascular endothelial growth factor (VEGF) mRNA levels in response to punicalagin treatment compared with control groups: L1, control; L2, L-NAME; L3, L-NAME+punicalagin (25 mg/kg); L4, L-NAME+punicalagin (50 mg/kg); L5, L-NAME+punicalagin (100 mg/kg). Values are expressed as means±SD, n=6; one-way ANOVA followed by DMRT showed p<0.05. Relative expression of proteins where the control expression level is set as 100%; *p<0.05 vs. control; #p<0.05 vs. L-NAME; the letters a–e indicate mean values from experimental groups that differ significantly, p<0.05.

• 

  Fig. 5 Punicalagin restored angiogenic balance.Western blotting analysis (a) and protein expression (b) revealed a marked decrease in VEGF levels in response to punicalagin treatment compared with control groups: L1, control; L2, L-NAME; L3, L-NAME+punicalagin (25 mg/kg); L4, L-NAME+punicalagin (50 mg/kg); L5, L-NAME+punicalagin (100 mg/kg). Values are expressed as means±SD, n=6; one-way ANOVA followed by DMRT showed p<0.05. Relative expression of proteins where the control expression level is set as 100%; *p<0.05 vs. control; #p<0.05 vs. L-NAME; the letters a–e indicate mean values from experimental groups that differ significantly, p<0.05.

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