Decreased body-fat accumulation and increased vasorelaxation to glyceryl trinitrate in middle-aged male rats following six-weeks consumption of coconut milk protein

Naphatthalung, Jomkarn; Chairuk, Pilaipan; Yorsin, Somruedee; Kanokwiroon, Kanyanatt; Radenahmad, Nisaudah; Jansakul, Chaweewan

doi:10.1590/s2175-97902022e20510

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Brazilian Journal of Pharmaceutical Sciences

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Article • Braz. J. Pharm. Sci. 58 • 2022 • https://doi.org/10.1590/s2175-97902022e20510 copy

Decreased body-fat accumulation and increased vasorelaxation to glyceryl trinitrate in middle-aged male rats following six-weeks consumption of coconut milk protein

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

We investigated whether coconut milk protein (CMP) contributes to the beneficial effects of coconut milk consumption on cardiovascular health markers previously found in middle-aged rats. CMP was isolated and precipitated from dried fresh coconut milk, then gavaged (1 g/kg) to middle-aged male rats for six weeks; control rats received distilled water. Compared to controls, CMP caused decreased body fat and lipid accumulation in liver cells and the platelet count. CMP did not affect basal blood pressure or heart rate in anesthetized rats. Vascular responsiveness to phenylephrine, DL-propargylglycine (PAG), acetylcholine or sodium nitroprusside was unaffected, but vasorelaxation to glyceryl trinitrate (GTN) increased. Effects of ODQ on vasorelaxation to GTN were similar in both groups. Expression of blood vessel eNOS, CSE and sGC was normal. The cyclic guanosine monophosphate (cGMP) level of CMP-treated rats was normal but addition of GTN increased cGMP and NO concentration more in CMP-treated rats than in controls, an effect unaltered by addition of diadzin. Taken together, CMP appears partially responsible for the improvement in cardiovascular health markers caused by coconut milk in middle-aged male rats.

Keywords:
Coconut protein; Blood vessel; Liver lipid; NO; Glyceryl trinitrate

NLAC-MU	Glucose	Triglyceride	Cholesterol	HDL-C	LDL-C	LDL/HDL
normal range						ratio
(mg %)	122.1-180.8	61.0-164.0	46.0-98.0	-	-	-
DW	132.2 ± 5.2	78.2 ± 7.9	95.6 ± 2.6	99.0 ± 5.5	25.3 ± 2.0	0.3 ± 0.03
CMP	131.0 ± 5.9	84.2 ± 11.2	90.8 ± 7.7	98.7 ± 5.6	23.3 ± 3.5	0.2 ± 0.05

NLAC-MU	ALP (U/L)	SGOT (U/L)	SGPT (U/L)	BUN (mg %)	CREAT (mg %)
normal range	46.0 - 92.0	111.0 - 225.0	25.0 - 64.0	10.3 - 23.6	0.5 - 0.7
DW	74.4 ± 7.8	123.9 ± 12.1	80.7 ± 5.6	23.8 ± 2.3	0.6 ± 0.1
CMP	71.4 ± 4.3	120.7 ± 11.4	77.6 ± 3.5	25.7 ± 3.9	0.7 ± 0.2

NLAC-MU	n	HCT (%)	HGB (g/dL)	MCV (fL)	MCH (pg)	MCHC (%)	WBC (x10³/µL)	Neutrophil (%)	Lymph (%)	Plt (x10⁵/µL)	N/L ratio
normal range		33.2 - 46.0	13.5 - 17.6	47.5 - 54.7	17.4 - 26.5	34.7 - 51.8	3.0 - 7.2	-	59.0 - 91.0	4.9 - 11.3	-
DW	6	46.8 ± 2.7	16.0 ± 0.8	51.6 ± 0.2	17.4 ± 0.2	33.4 ± 0.7	5.5 ± 0.6	68.2 ± 6.1	31.4 ± 6.1	8.6 ± 0.3	2.7 ± 0.6
CMP	6	46.1 ± 2.3	16.2 ± 0.7	52.9 ± 1.2	18.4 ± 0.4	34.7 ± 0.4	4.4 ± 0.3	63.3 ± 4.8	35.4 ± 4.9	7.4 ± 0.6^a	2.1 ± 0.4

Treatments	Adipose tissue weight/100 g body weight (% g)
Treatments	Epididymis	Prostate	Mesentery	Retroperitoneal	Subcutaneous
DW	2.48 ± 0.15	0.18 ± 0.03	2.43 ± 0.20	3.23 ± 0.58	7.70 ± 0.68
CMP	2.22 ± 0.06	0.19 ± 0.00	1.78 ± 0.14^a	1.97 ± 0.29^a	5.28 ± 0.68^a

Treatments	Organs weight/ 100 g body weight (% g)
Treatments	Heart	Lung	Liver	Kidney	Adrenal gland (mg)	Spleen	Testis	Epididymis	Prostate gl.
DW	0.3 ±	0.3 ±	2.7 ±	0.5 ±	13.4 ±	0.2 ±	0.6 ±	0.3 ±	0.2 ±
	0.01	0.01	0.15	0.02	0.99	0.01	0.01	0.01	0.02
CMP	0.3 ±	0.3 ±	2.7 ±	0.6 ±	14.3 ±	0.2 ±	0.6 ±	0.3 ±	0.2 ±
	0.01	0.01	0.16	0.06	0.71	0.01	0.01	0.01	0.02

Treatments	Body weight		n	Basal	Basal	Mean Arterial	Basal
Treatments	(g)			systolic BP	diastolic BP	Pressure	heart rate
	Initial	Final		(mmHg)	(mmHg)	(mmHg)	(bpm)
DW	481.9 ± 13.1	492.1 ± 8.4	6	134.2 ± 5.4	109.2 ± 4.6	120.8 ± 2.0	425.0 ± 11.2
CMP	488.0 ± 7.9	505.1 ± 7.2	6	136.7 ± 9.6	109.6 ± 8.0	118.6 ± 8.5	428.3 ± 10.9

	EC₅₀ (nM) : 95% confidential limit		E_max (g)
	DW	CMP	DW	CMP
Phenylephrine
Endo	14.0 (7.8-25.2)	25.2 (16.0-39.7)	3.6 ± 0.3	2.9 ± 0.3
Endo+L-NA	19.3 (13.3-31.2)	24.0 (15.7-37.0)	7.1 ± 0.6	6.5 ± 0.4
Endo+L-NA+PAG	67.6 (34.6-98.0)	30.1 (19.2-49.7)	6.8 ± 0.6	7.5 ± 0.5
No endo	26.2 (8.6-79.6)	33.7 (14.7-77.0)	4.7 ± 0.3	4.8 ± 0.2
No endo+L-NA	21.1 (9.2-48.4)	36.4 (15.6-84.9)	6.7 ± 0.4	6.9 ± 0.1
No endo+L-NA+PAG	21.7 (10.7-44.2)	11.3 (6.3-20.3)	8.3 ± 0.6	8.2 ± 0.2

	EC50 (nM) : 95% confidential limit		E_max (g)
	DW	CMP	DW	CMP
Acetylcholine
Endo	53.6 (5.1-268.0)	62.7 (5.3-297.5)	64.6 ± 4.6	68.8 ± 6.3
Endo+PAG	72.0 (9.8-329.0)	128.9 (10.0-366.9)	45.7 ± 3.0	50.2 ± 3.2
Sodium nitroprusside
Endo	13.5 (9.9-18.2)	11.9 (10.8-13.1)	119.0 ± 4.7	125.4 ± 2.9
Endo+L-NA	3.7 (2.6-5.2)	3.3 (2.2-5.1)	108.1 ± 2.5	111.2 ± 1.8
Endo+L-NA+PAG	61.3(55.4-67.9)	59.8(55.8-64.1)	95.1 ± 2.9	94.9 ± 1.8
Glyceryl trinitrate
Endo	64.4 (32.6-117.5)	35.0 (29.1-42.0)	68.1 ± 4.1	93.8 ± 4.1a
Endo+L-NA	48.7 (28.1-84.3)	27.4 (11.3-66.7)	95.9 ± 3.4	105.6 ± 2.1^a
Endo+L-NA+PAG	50.5 (34.8-73.3)	27.4 (9.6-78.6)	89.4 ± 2.7	100.0 ± 1.3^a

Universidade de São Paulo, Faculdade de Ciências Farmacêuticas Av. Prof. Lineu Prestes, n. 580, 05508-000 S. Paulo/SP Brasil, Tel.: (55 11) 3091-3824 - São Paulo - SP - Brazil
E-mail: bjps@usp.br

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[1] *Correspondence: C. Jansakul. Faculty of Traditional Thai Medicine. Prince of Songkla University. Hat-Yai, Thailand 90110. Phone: 66-074-286824. E-mail address: chaweewan.j@psu.ac.th.