Caffeine suppresses the expression of the Bcl-2 mRNA in BeWo cell culture and rat placenta.

Chronic caffeine exposure during pregnancy has an effect on fetal growth; however, the adverse effects of caffeine on embryogenesis are not well understood and controversial. We used cDNA microarray technology to determine whether caffeine alters gene expressions in a human cytotrophoblast-like cell line, BeWo. We found that the expression of the B-cell CLL/lymphoma 2 (Bcl-2) gene in BeWo cells was down-regulated by caffeine, suggesting that chronic exposure during the gestational period could exert an influence on embryogenesis. We then focused on the Bcl-2- and Bcl-2-associated X protein gene, Bax, to study the responsive gene expression in BeWo cells as well as placentas of pregnant rats fed a diet supplemented with caffeine (2 mg/100 g body weight) during gestation, and analyzed the gene expressions using LightCycler-based quantitative real-time polymerase chain reaction assays. We found a significantly decreased level of Bcl-2 mRNA expression, which demonstrated the influence of caffeine on placental function.

Effects of caffeine on the saturated and monounsaturated Fatty acids of the newborn rat cerebellum.

Caffeine (1,3,7-trimethylxanthine) is one of the most commonly consumed drugs in our daily life, and its use is increasing. However, very little attention has been paid to its potential effects on early growth and development. Because of the steady increase in breast feeding of infants and because caffeine diffuses readily into breast milk, the present study examined if caffeine intake by newborn rats during lactation would affect the saturated and monounsaturated fatty acids in the growing cerebellum. A total of 10 timed pregnant rats were purchased from the breeder. At birth litters were combined, and 8 pups were randomly assigned to each dam without regard to the sex of the pups. Dams with litters were divided into 2 groups. Dams of group 1 received a 20% protein diet as a control, and dams of group 2 received a 20% protein diet plus caffeine (4 mg/100 g BW). Pups were killed at day 10. The cerebellums were removed, weighed and homogenized. Gas chromatograph-mass spectrometry was used to identify and quantify free fatty acids. Chronic caffeine exposure from birth to day 10 in pups through the maternal milk resulted in a decrease in cerebellum weight, a significant increase in the saturated fatty acids, and a tendency toward an increase of monounsaturated fatty acids. In addition, there was a slight increase of some of the polyunsaturated fatty acids. However, there was no difference in food intake of the lactating dams and weight gain of the pups between the groups. These data indicate that early caffeine intake by the suckling pups alters the composition of fatty acids of the cerebellum; thus, avoidance of caffeine during lactation is critical. The risks and benefits of caffeine administration in premature infants must be carefully evaluated during this rapid period of brain growth.

A caffeine diet can alter the mechanical properties of the bones of young ovariectomized rats.

The general public widely consumes caffeine which is contained in various foods, beverages, and over-the-counter medications. The relationships between caffeine intake and bone fractures is controversial. Therefore, the aim of the current study was to determine what effects, if any, caffeine intake in early life exerts on mechanical properties and mineral contents of bone in growing ovariectomized rats. A total of 8 dams with pups were divided into two groups. Group 1 was fed a 20% protein diet. Group 2 was fed a 20% protein diet supplemented with caffeine (4 mg/100 g). The respective diets were fed to the dams during lactation and to the pups continuously after weaning on day 22 until the end of the experimental period. On day 32, offspring from both groups were ovariectomized. On day 52, the rats were sacrificed and the femora removed. The biomechanical properties of the femora were determined by three-point bend testing to failure at a rate of 2 mm/min, with continuous data sampling at 10 samples/s. The properties determined included the modulus of elasticity, yield load, yield stress, ultimate load, ultimate stress, and the second moment of area. The caffeine group exhibited a decrease in the various mechanical properties (ranging from approximately 7 to 20%), except for yield strain and moment of inertia. The decreases in maximum stress and elastic modulus values were significant. Calcium, magnesium, and phosphorous values for the caffeine group were significantly decreased. These results suggest that the bone in the caffeine group is weaker and less stiff, with greater deformation under applied loading. It could be concluded that caffeine intake during the early growing period affects the mechanical properties of bone.

Association of different zinc concentrations combined with a fixed caffeine dose on plasma and tissue caffeine and zinc levels in the rat.

Because caffeine and tissue levels of Zn are closely related, the objectives of this study were to determine the changes in plasma caffeine levels over a period of 5 h when different concentrations of Zn combined with a fixed concentration of caffeine were injected into the femoral vein of rats and to determine the relationship between tissue levels of caffeine and Zn at 5 h postinjection. Rats were divided into three groups: group 1, 220 microg caffeine; group 2, 220 microg caffeine + 8 microg Zn/g body weight (BW); group 3, 220 microg caffeine + 16 microg Zn/g BW. Blood from groups 1 and 3 was collected at 3 min, 30 min, 1 h, 3 h, and 5 h to determine the pharmacokinetics of caffeine. All groups were killed at 5 h. Caffeine and Zn concentrations of the brain, kidney, heart, and liver of all groups were determined. The plasma-caffeine curve in group 3 showed a lower concentration at 3 min and a slower caffeine-elimination rate during the first 3 h. Brain and kidney caffeine levels remained constant in all groups, whereas caffeine levels were increased in the heart in group 2 and in the liver in group 3. Zn concentrations in the brain and kidney were lower in group 2 compared with groups 1 and 3 and higher in group 3 compared to groups 1 and 2. Zn concentration in the heart was the same among the three groups but was increased in the liver in group 3 compared to groups 1 and 2. Therefore, we concluded that caffeine combined with Zn affects caffeine pharmacokinetics. With caffeine intake, levels of Zn (16 microg/g BW) that are slightly higher than the daily requirements (12 microg/g BW) may prevent a reduction of Zn in tissue. In addition, caffeine's effects on Zn concentration among organs are different.