Gut Microbiota Perturbation in Early Life Could Influence Pediatric Blood Pressure Regulation in a Sex-Dependent Manner in Juvenile Rats

Abstract

The present study aimed to investigate whether gut dysbiosis induced by ceftriaxone in early life could influence pediatric blood pressure regulation in childhood with or without exposure to a high-fat diet (HFD). Sixty-three newborn pups of Sprague-Dawley rats were administered ceftriaxone sodium or saline solution until weaning at 3 weeks, and the rats were fed a HFD or regular diet from 3 to 6 weeks. Tail-cuff blood pressure, the expression levels of genes of the renin-angiotensin system (RAS), the concentrations of IL-1β, IL-6, and TNF-α in the colon and prefrontal cortex, and the composition of fecal microbiota were analyzed. Ceftriaxone treatment significantly increased the diastolic blood pressure of male rats at 3 weeks. At 6 weeks, systolic blood pressure (SBP) was significantly increased only in ceftriaxone treated male rats fed with HFD. The RAS showed increased activation in the kidney, heart, hypothalamus, and thoracic and abdominal aorta of male rats, but only in the kidney, heart, and hypothalamus of female rats. HFD-fed female rats showed a decreased level of IL-6 in the colon. α diversity of gut microbiota decreased and the Firmicutes to Bacteroidetes ratio increased in both male and female rats at 3 weeks; however, these parameters recovered to various degrees in female rats at 6 weeks. These results revealed that early-life gut dysbiosis induced by antibiotics combined with a HFD in childhood could be involved in pediatric blood pressure regulation and an increase in SBP in juvenile rats, and these effects occurred in a sex-dependent manner.

Keywords: blood pressure; ceftriaxone; gut microbiota; high-fat diet; pediatric hypertension.

Figure 1

Body weight of (A) 0–3 weeks (Control n = 31, CEF n = 32), (B) 3–6 weeks of male rat (Control n = 6, CEF n = 7, HFD n = 8, CEF + HFD n = 7), and (C) 3–6 weeks of female rats (Control n = 9, CEF n = 9, HFD n = 8, CEF + HFD n = 9). CEF, ceftriaxone group. HFD, high-fat diet group. CEF + HFD, ceftriaxone followed with high-fat diet group. PND: postnatal day. * p < 0.05 between Control vs. CEF.

Figure 2

Systolic blood pressure (SBP) and diastolic blood pressure (DBP) of the rats. (A,B) male rats at the age of 3 weeks (Control n = 8, CEF n = 12), (C,D) female rats at the age of 3 weeks (Control n = 9, CEF n = 15), (E,F) male rats at the age of 6 weeks (Control n = 6, CEF n = 7, HFD n = 8, CEF + HFD n = 7), (G,H) female rats at the age of 6 weeks (Control n = 9, CEF n = 9, HFD n = 8, CEF + HFD n = 9). CEF ceftriaxone group, HFD high-fat diet group, CEF + HFD ceftriaxone and high-fat diet group. * p < 0.05, ** p < 0.01.

Figure 3

Concentrations of cytokines in colon and prefrontal cortex of (A,B) male rats at 6 weeks of age (Control n = 6, CEF n = 7, HFD n = 8, CEF + HFD n = 7) and (C,D) female rats at 6 weeks of age (Control n = 9, CEF n = 9, HFD n = 8, CEF + HFD n = 9). CEF, ceftriaxone group. HFD, high-fat diet group. CEF + HFD, ceftriaxone and high-fat diet group. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Figure 4

α diversity and principal coordinate analysis (PCoA) of fecal microbiota at the age of 3 and 6 weeks in male rats. (A–C) α diversity at 3 weeks of age (Control n = 6, CEF n = 6). (D) PCoA of fecal microbiota at 3 weeks of age. (E–G) α diversity at 6 weeks of age (Control n = 6, CEF n = 7, HFD n = 8, CEF + HFD n = 7). (H) PCoA of fecal microbiota at 6 weeks of age. CEF, ceftriaxone group. HFD, high-fat diet group. CEF + HFD, ceftriaxone and high-fat diet group. * p < 0.05, ** p < 0.01, *** p < 0.001.

Figure 5

α diversity and principle coordinate analysis (PCoA) of fecal microbiota at the age of 3 and 6 weeks of female rats. (A–C) α diversity at 3 weeks of age (Control n = 6, CEF n = 6). (D) PCoA of fecal microbiota at 3 weeks of age. (E–G) α diversity at 6 weeks of age (Control n = 9, CEF n = 9, HFD n = 8, CEF + HFD n = 9). (H) PCoA of fecal microbiota at 6 weeks of age. CEF, ceftriaxone group. HFD, high-fat diet group. CEF + HFD, ceftriaxone and high-fat diet group. ** p < 0.01, *** p < 0.001.

Figure 6

Relative abundance and heat map of gut microbiota for the top 10 categories of gut microbiota in (A1–A4) male rats at 3 weeks of age (Control n = 6, CEF n = 6), (B1–B4) male rats at 6 weeks of age (Control n = 6, CEF n = 7, HFD n = 8, CEF + HFD n = 7), (C1–C4) female rats at 3 weeks of age (Control n = 6, CEF n = 6), and (D1–D4) female rats at 6 weeks of age (Control n = 9, CEF n = 9, HFD n = 8, CEF + HFD n = 9). CEF, ceftriaxone group. HFD, high-fat diet group. CEF + HFD, ceftriaxone and high-fat diet group. * p < 0.05 and ** p < 0.01 for data of 3 weeks. No common small letter means p < 0.05 and no common capital letter means p < 0.01 for data of 6 weeks.

Figure 7

Concentrations of SCFAs of feces in (A) male rats at 3 weeks of age (Control n = 6, CEF n = 6), (B) male rats at 6 weeks of age (Control n = 6, CEF n = 7, HFD n = 8, CEF + HFD n = 7), (C) female rats at 3 weeks of age (Control n = 6, CEF n = 6), and (D) female rats at 6 weeks of age (Control n = 9, CEF n = 9, HFD n = 8, CEF + HFD n = 9). CEF, ceftriaxone group. HFD, high-fat diet group. CEF + HFD, ceftriaxone and high-fat diet group. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Figure 8

Concentrations of serum TMAO and its relative components in (A) male rats (Control n = 6, CEF n = 7, HFD n = 8, CEF + HFD n = 7) and (B) female rats (Control n = 9, CEF n = 9, HFD n = 8, CEF + HFD n = 9). CEF, ceftriaxone group. HFD, high-fat diet group. CEF + HFD, ceftriaxone and high-fat diet group. * p < 0.05, ** p < 0.01, *** p < 0.001 **** p < 0.0001.

Figure 9

Spearman rank correlative matrix between SBP/DBP and biochemical indicators including gut microbiota, SCFAs, TMAO and precursors, and cytokines in (A) male rats and (B) female rats. SBP, systolic blood pressure. DBP, diastolic blood pressure. TMA, trimethylamine, TMAO, trimethylamine N-oxide. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.