Lactoferrin Supplementation during Pregnancy and Lactation Protects Adult Male Rat Offspring from Hypertension Induced by Maternal Adenine Diet

Abstract

Lactoferrin, a glycoprotein derived from breastmilk, is recognized for its health benefits in infants and children; however, its protective effects when administered during gestation and lactation against offspring hypertension remain unclear. This study aimed to investigate whether maternal lactoferrin supplementation could prevent hypertension in offspring born to mothers with chronic kidney disease (CKD), with a focus on nitric oxide (NO), renin-angiotensin system (RAS) regulation, and alterations in gut microbiota and short-chain fatty acids (SCFAs). Prior to pregnancy, female rats were subjected to a 0.5% adenine diet for 3 weeks to induce CKD. During pregnancy and lactation, pregnant rats received one of four diets: normal chow, 0.5% adenine diet, 10% lactoferrin diet, or adenine diet supplemented with lactoferrin. Male offspring were euthanized at 12 weeks of age (n = 8 per group). Supplementation with lactoferrin during gestation and lactation prevented hypertension in adult offspring induced by a maternal adenine diet. The maternal adenine diet caused a decrease in the index of NO availability, which was restored by 67% with maternal LF supplementation. Additionally, LF was related to the regulation of the RAS, as evidenced by a reduced renal expression of renin and the angiotensin II type 1 receptor. Combined maternal adenine and LF diets altered beta diversity, shifted the offspring's gut microbiota, decreased propionate levels, and reduced the renal expression of SCFA receptors. The beneficial effects of lactoferrin are likely mediated through enhanced NO availability, rebalancing the RAS, and alterations in gut microbiota composition and SCFAs. Our findings suggest that maternal lactoferrin supplementation improves hypertension in offspring in a model of adenine-induced CKD, bringing us closer to potentially translating lactoferrin supplementation clinically for children born to mothers with CKD.

Keywords: chronic kidney disease; developmental origins of health and disease (DOHaD); gut microbiota; hypertension; lactoferrin; nitric oxide; renin–angiotensin system.

Figure 1

Effects of maternal adenine diet (CKD) and lactoferrin (LF) on systolic blood pressure in offspring from Week 3 to 12. N = 8/group. * p < 0.05 vs. CN; # p < 0.05 vs. CKD.

Figure 2

Effects of maternal adenine diet (CKD) and lactoferrin (LF) on the renin–angiotensin system at Week 12. N = 8/group. * p < 0.05 vs. CN.

Figure 3

The evaluation of gut microbial biodiversity in offspring born to dams fed an adenine (CKD) or lactoferrin (LF) diet. (A) Faith’s phylogenic diversity (pd), (B) Shannon index, and (C) principal coordinate analysis (PCoA). Outliers are denoted by dots. Each color corresponds to a different group, with each data point representing one sample.

Figure 4

Linear discriminant analysis effect size (LEfSe) with an LDA score > 4 identified significantly differential taxa between groups. The respective group is denoted by the color of the horizontal bar.

Figure 5

Bar plots showing the genus-level discrimination between the CKD and CKDLF groups.

Figure 6

The comparison of relative abundance of genus Robinsoniella among the four groups. Outliers are denoted by dots. ** p < 0.01. **** p < 0.001.

Figure 7

Plasma concentrations of (A) acetate, (B) propionate, and (C) butyrate, and (D) renal mRNA expression of SCFA receptors at Week 12. N = 8/group. * p < 0.05 vs. CN.