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.2022 Jul 28;17(7):e0271735.
doi: 10.1371/journal.pone.0271735. eCollection 2022.

Aqueous extracts of Urtica dioica (stinging nettle) leaf contain a P2-purinoceptor antagonist-Implications for male fertility

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Aqueous extracts of Urtica dioica (stinging nettle) leaf contain a P2-purinoceptor antagonist-Implications for male fertility

Nicole T Eise et al. PLoS One..

Abstract

Stinging nettle root and leaf extracts were tested for their effect on prostatic smooth muscle contractility. Root extract did not affect electrical field stimulation induced-nerve mediated contractions of isolated rat prostates. On the other hand, leaf extract attenuated electrical field stimulation-induced contractions at all frequencies. Similarly, contractions elicited by exogenous administration of ATP and αβ-methylene ATP were inhibited by leaf extract, whereas contractions elicited by exogenous administration of noradrenaline or acetylcholine were unaffected. The active component was present within the aqueous phase of the leaf extract. In mouse mating studies, stinging nettle leaf extract (50 mg p.o. daily) reduced male fertility by 53% compared to vehicle-treated male mice. Cardiovascular parameters were unaffected by administration of stinging nettle leaf extract (p ≥ 0.057). Treated mice exhibited normal mating behaviour. Bladder and testes weighed less in stinging nettle leaf extract treated mice. All other organs and total body weight were unaffected. It is concluded that stinging nettle leaf extract reduces contractility of genitourinary smooth muscle by acting as an antagonist at postjunctional P2X1-purinoceptors. These data indicates that blocking sperm transport through pharmacological blockade of P2X1-purinoceptors via oral administration is consistent with an effective and convenient biological strategy male contraception.

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Conflict of interest statement

The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1. Effect of stinging nettle leaf extract on prostate smooth muscle contractility.
Mean contractile responses to electrical field stimulation (0.5 ms, 60 V, 0.1 Hz to 20 Hz for 10 pulses or 10 seconds) of isolated rat prostate gland preparations in the presence of (A) vehicle (ethanol 0.25% v/v) and stinging nettle leaf extract (5 mg/ml) or (B) vehicle (ethanol 0.75% v/v) and stinging nettle leaf extract (15 mg/ml). Mean contractile responses to exogenously administered (C) noradrenaline (1 nM– 30 μM), (D) acetylcholine (1 nM– 100 μM), (E) adenosine 5’-triphosphate (ATP: 300 nM– 1 mM) and (F) α,β-methylene ATP (3 nM– 10 μM) of isolated rat prostate gland preparations in the presence of [○] vehicle (ethanol 0.25% or 0.75% v/v) and [□] stinging nettle leaf extract (5 or 15 mg/ml). In all graphs, each column or point represents the mean force generated by prostates taken from six rats. Error bars represent SEM. ANOVAp-values were determined by two-way repeated-measures ANOVA and represent the probability that the observed differences were due to chance.
Fig 2
Fig 2. Effect of aqueous and organic fractions of stinging nettle leaf extract on smooth muscle contractions.
Mean contractile responses to EFS (0.5 ms, 60 V, 0.1 Hz to 1 Hz for 10 pulses) of isolated rat prostate gland preparations in the presence of (A) vehicle (ethanol 0.25% v/v) and the organic fraction of stinging nettle leaf extract (0.035 mg/ml) or (B) vehicle (ethanol 0.25% v/v) and the aqueous fraction of stinging nettle leaf extract (2.09 mg/ml). Columns represent the mean force generated by prostates taken from six rats. Error bars represent SEM; and (C) mean contractile responses to exogenously administered α,β-methylene ATP (3 nM– 10 μM) of isolated rat prostate gland preparations in the presence of [○] vehicle (ethanol 0.25%) and [▲] the aqueous fraction of stinging nettle leaf extract (2.09 mg/ml, lower panel). Each point represents the mean force generated by prostates taken from six rats. Error bars represent SEM. In all graphsp-values were determined by two-way repeated-measures ANOVA and represent the probability that the observed differences were due to chance.
Fig 3
Fig 3. Effect of stinging nettle leaf extract or P2X1-purinoceptor deletion on resting cardiovascular parameters and male fertility.
Mean resting (A) systolic blood pressure (mmHg) and (B) heart rate (bpm) measured by the tail cuff method in untreated age matched wild-type (n = 6) and P2X1-purinoceptor knockout (KO) (n = 6), and wild-type male mice, before and after daily oral dosing with stinging nettle leaf extract (SNLE) (100 μl of 500 mg/ml) or vehicle (100 μl of 20% ethanol), where n = 6 for each treatment variable. Columns represent the mean systolic blood pressure (A) or heart rate (B) over 5 consecutive days prior to the commencement of treatment or 5 consecutive days after the commencement of treatment. Error bars represent SEM. ANOVAp-values were determined by two-way repeated-measures ANOVA and represent the probability of genotype or treatment causing a significant change in systolic blood pressure or heart rate. (C) Foetal number resulting from wild-type female mice matings with wild-type male mice treated with vehicle (100 μl of 25% ethanol) (●), an oral daily dosing of stinging nettle leaf extract (SNLE) (100 μl of 500 mg/ml) (○) or age matched P2X1-purinoceptor knockout male mice (Δ). Six male mice were each mated with two female wild type mice for each treatment variable. The centre line represents the mean foetal number. Error bars represent SEM. ANOVAp-values were determined by one-way ANOVA and represent the probability of genotype or treatment affecting male fertility represented by the resultant foetal number.
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