## 4 Discussion  
The current investigation demonstrated that long-term dietary hempseed enrichment at 5% and 15% concentrations during early life does not reduce adult blood pressure, but a 15% dose may impact the pattern of blood pressure changes seen in growing mice fed a control diet. The findings presented here comprise the first report on the effect of habitual hempseed consumption on changes in blood pressure during growth and development. Figure 4: Box plots for systolic (blue), diastolic (red), and mean arterial (green) blood pressure. The line within the box indicates the median value; the upper edge of the box indicates the upper quartile; the lower edge of the box indicates the lower quartile; the bars extending beyond the box indicate the maximum and minimum values; the single points indicate outliers. (**A**) Control diet; (**B**) 5% hempseed; (**C**) 15% hempseed. \(n=8\) per diet group. \(p\) values are for the ANOVA main effect of age on blood pressure parameter within diet group. \({}^{*}\) indicates significant difference (\(p<0.05\)) compared to age of 5 weeks by [PERSON]’s post hoc analysis. The mice in this study were fed a hempseed-enriched diet from shortly past weaning (age 5 weeks) to mid-adulthood (age 30 weeks), a period encompassing blood pressure maturation and stabilization prior to late adulthood. Young developing mice showed a pattern of change in systolic, diastolic, and mean arterial blood pressure across the 25 weeks of this study characterized by a gradual rise from age 5 to ~11-17 weeks followed by a slow decline until age 30 weeks, the end of the experiment. Our findings are consistent with those of [PERSON] et al., who reported developmental increases in systolic and diastolic blood pressure in C57 BL/6 mice from post-natal ages of 21-50 days followed by relatively constant levels [21]. Our study and [PERSON] et al. 's study collected blood pressures measurements at different ages, rendering the results not directly comparable, but the pattern of blood pressure maturation was similar between the experiments. Similarly, [PERSON] et al. reported significantly higher systolic blood pressure in C57 BL/6 mice at age 24 weeks compared to 8 weeks [33]. [PERSON] et al. [34] observed increases in left ventricular mass in C57 BL/6 mice between the ages of 3 days to 4 months, which could be expected to parallel increases in systolic blood pressure [35]. Hempseeds contain molecules such as polyphenols, peptides, fatty acids, and micronutrients that exert biological activity in the body [36; 37]. Mechanisms underlying the effect of hempseed on blood pressure might involve the renin-angiotensin-aldosterone system and antioxidant defense and endocrine networks. Hempseed products, such as hempseed protein hydrolysate, demonstrate inhibitory actions on renin and angiotensin-converting enzyme activity in in vitro studies and cause reductions in systolic blood pressure in rats [15; 38]. Hempseed protein short- and medium-chain peptides have also been shown to inhibit dipeptidyl peptidase IV (DPP-IV), which degrades glucagon-like peptide-1 (GLP-1) and thereby modulates the renin-angiotensin system that regulates blood pressure [39; 40; 41; 42]. Hempseed peptides and lipids also support antioxidant defenses by scavenging free radicals [37; 43; 44], which modulate nitric oxide activity and vascular function [45; 46]. Hempseed-enriched diets also affect estradiol levels in rats [47], and estradiol modulates blood pressure in women [48]. Moreover, during puberty in humans, multiple physiological pathways impact the development of adult blood pressure [49], and these may be affected by bioactive components in hempseed. The apparent blunting of blood pressure changes across age in the 15% HS diet group relative to the control and 5% HS groups, and the delay in peak blood pressure in the 5% HS versus control diet group, might be worthy of further investigation.