In order to evaluate the serum lipid profile and the productive performance of laying hens at 45 weeks of age, 160 White Leghorn (L33) laying hens were allotted during 91 d, according to completely randomized design, into four treatments and 20 repetitions, at the time of laying peak. The treatments consisted of diets containing: 0, 3.3, 6.6, and 10% pumpkin seed meal (PSM) in the feedstuff. The viability, the laying intensity, the mass conversion and the egg weight did not differ significantly between treatments. The concentration of triacylglycerids, total cholesterol, low-density lipoproteins (LDL) and very-low-density lipoproteins (VLDL) disminished in 21.7, 33.7, 16.3, and 55.3 mg/dL, respectively, with 10% of PSM in the feedstuff, compared to the control. Moreover, the atherogenic index decreased from 3.33 to 2.96 and the high-density lipoproteins (HDL) increased, from 47.46 to 49.92 mg/dL, when including in the feedstuff up to 10% of PSM. The octadecanoic (C18:0), oleic (C18: 1n9), linoleic (C18:2n6), and α-linolenic (C 18:3n3) acids were increased in 66.79, 21.60, 57.73, and 20.10 mg/dL, respectively; whereas the arachidonic acid (C20:4n6) declined to 6.69 mg/dL, with 10% of PSM in the feedstuff, compared to the control. It is recommended to use up to 10% of pumpkin seed meal in laying hen diets to substitute plant oil and imported soybean cake, reduce harmful lipids, and increase the essential fatty acids circulating in the blood serum, not harming the productive performance of the hens.