<p>Preschool is a critical time to establish healthy eating and physical activity (PA) habits, yet most preschoolers and their caregivers do not consume overall healthy diets or meet PA recommendations. This before and after study assessed whether a multi-level intervention using the social ecological model, the Head Start to Healthy Lifestyles (HSHL) project, improved dietary intake and PA among Head Start preschoolers and their caregivers. In year 2 of the project, HSHL was integrated into 8 Head Start preschool centers in Denton and Tarrant Counties in North Texas, U.S.A, serving 460 preschool-aged children and their caregivers. Weekly, 30-minute nutrition and PA activities were delivered to preschoolers in classrooms for 9 months. Caregivers received online nutrition and PA content. Fruit and vegetable intake, nutrition-related knowledge, PA, and sedentary behaviors were evaluated using McNemar and Wilcoxon’s tests. From 177 completed pre-surveys (12 duplicates) and 174 post-surveys (13 duplicates), 44 matched surveys were identified. Caregivers were predominantly female (95%), and aged 18–59 years. Children were 3–5 years old. Children decreased drinking fruit juice (<i>p</i> = 0.044) and increased engaging in PA (<i>p</i> = 0.001). There were increases in caregivers’ fruit consumption (<i>p</i> = 0.023), use of MyPlate to plan meals (<i>p</i> = 0.006), and use of food labels (<i>p</i> = 0.005). The findings suggest that educational interventions may promote healthier lifestyle behaviors in Head Start preschoolers and caregivers.</p>