Many studies have shown that there is a relationship between gonadotropin-releasing hormone (GnRH) and glucose metabolism, but little is known about the effects of GnRH on the pancreas. Our experiment investigated the effect of GnRH on pancreatic islet cell in Sprague–Dawley (SD) rats fed with high-cholesterol diet before and during pregnancy. We found that although high-cholesterol diet led to no significant difference of GnRH mRNA levels in pancreas in nonpregnant rats, it led to a marked increase of those in pregnant rats. Furthermore, in rats fed with standard laboratory chow, no significant differences were apparent in GnRH mRNA levels before and during gestation; however, when fed with high-cholesterol diet, the GnRH mRNA levels increased significantly in pregnant rats. As results indicated both diets could lead to increase of PG mRNA in pancreas of pregnant rats. It is also demonstrated that the GnRH mRNA levels are positively associated with PG mRNA levels. Moreover, our data showed a significant increase in fasting insulin level in the Gestation group compared with Control. Such changes were contrary to the changes of GnRH level in the pancreas. This may imply that GnRH influences hormones secretion in the pancreas by autocrine and paracrine effects on islet cells.