Ethyl Acetate Fraction from Leandra dasytricha (A. Gray) Cong. Leaves Promotes Vasodilatation and Reduces Blood Pressure in Normotensive and Hypertensive Rats

Abstract

Leandra dasytricha (A. Gray) Cong. is widely distributed in the south of Brazil and is commonly used for cardiovascular and kidney ailments. For this study, we used male Wistar normotensive rats (NTRs) and spontaneously hypertensive rats (SHRs) to verify the effects of the ethyl acetate fraction (EAF) obtained from L. dasytricha leaves on isolated aorta relaxation and in the arterial blood pressure. The EAF was analyzed by LC-DAD-MS, and several components were annotated, including hydrolysable tannins, triterpenes, and O- and C-glycosylated dihydrochalcones, such as the most intense ion peak relative to C-hexosyl phloretin (nothofagin; compound number 13). The EAF caused a concentration and endothelium-dependent relaxation of the aorta in both NTRs and SHRs. This effect was abolished in the endothelium-denuded aorta. L-NAME, a nonselective nitric oxide synthase inhibitor, and ODQ, a soluble guanylate cyclase inhibitor, entirely blocked the EAF-induced relaxation. The presence of a muscarinic receptor antagonist or a cyclooxygenase inhibitor did not alter the EAF's effectiveness in relaxing the aorta. The preincubation with tetraethylammonium, a Ca²⁺-activated K⁺ channel blocker, and with 4-aminopyridine, a voltage-dependent K⁺ channel blocker, significantly interfered with the EAF's relaxation. However, the incubation with glibenclamide, an ATP-sensitive K⁺ channel blocker, and barium chloride, an inward-rectifier K⁺ channel blocker, did not interfere with the EAF-induced relaxation. The EAF treatment also caused a dose-dependent decrease in the mean arterial pressure, systolic arterial pressure, and diastolic arterial pressure of both NTRs and SHRs, without significantly interfering with heart rate values. In conclusion, this study demonstrated the EAF-induced vasorelaxant and hypotensive actions, primarily dependent on the endothelium function and mainly with the participation of the nitric oxide and Ca²⁺-activated and voltage-dependent K⁺ channels.

Figure 1

Base peak chromatograms obtained in positive (a) and negative ion mode (b) chromatograms from EAF.

Figure 2

Vasorelaxation induced by EAF in the aorta of Wistar normotensive rats (NTRs) and spontaneously hypertensive rats (SHRs). Concentration-response curves were determined in endothelium-intact (E+) or endothelium-denuded (E-) rings. (a) The EAF-induced vasorelaxation in the NTR aorta. (b) EAF-induced vasorelaxation in the SHR aorta. Statistical analyses were performed using a two-way analysis of variance followed by Bonferroni's multiple comparison test. ^∗p < 0.05 when compared to the vehicle (closed triangles), and ^#p < 0.05 when compared to E- (opened squares).

Figure 3

Vasorelaxation induced by EAF in the aorta of Wistar normotensive rats (NTRs) and spontaneously hypertensive rats (SHRs) in the absence or presence of L-NAME (a, b) and ODQ (c, d). Statistical analyses were performed using a two-way analysis of variance followed by Bonferroni's multiple comparison test. ^∗p < 0.05 when compared to the vehicle (closed triangles), and ^#p < 0.05 when compared to the vehicle-only incubation (opened squares).

Figure 4

Vasorelaxation induced by EAF in the aorta of Wistar normotensive rats (NTRs) and spontaneously hypertensive rats (SHRs) in the absence or presence of atropine (a, b) and indomethacin (c, d). Statistical analyses were performed using a two-way analysis of variance followed by Bonferroni's multiple comparison test. ^∗p < 0.05 when compared to the vehicle (closed triangles).

Figure 5

Vasorelaxation induced by EAF in the aorta of Wistar normotensive rats (NTRs) and spontaneously hypertensive rats (SHRs) in the absence or presence of TEA 10 mM (a, b) and TEA 1 mM (c, d). Statistical analyses were performed using a two-way analysis of variance followed by Bonferroni's multiple comparison test. ^∗p < 0.05 when compared to the vehicle (closed triangles), and ^#p < 0.05 when compared to the vehicle-only incubation (opened squares).

Figure 6

Vasorelaxation induced by EAF in the aorta of Wistar normotensive rats (NTRs) and spontaneously hypertensive rats (SHRs) in the absence or presence of 4-AP (a, b), glibenclamide (c, d), and BaCl₂ (e, and f). Statistical analyses were performed using a two-way analysis of variance followed by Bonferroni's multiple comparison test. ^∗p < 0.05 when compared to the vehicle (closed triangles), and ^#p < 0.05 when compared to the vehicle-only incubation (opened squares).