Abstract: Objective To observe the effect of Guben Zenggu Decoction on the expressions of calmodulin (CaM) and calcium/calmodulin-dependent protein kinase Ⅱ (CaMK Ⅱ) mRNA in the hypothalamus, pituitary, spleen, adrenal gland and femur in ovariectomized rats. To explore the mechanism of Guben Zenggu Decoction regulating and controlling osteoporosis through neuro-endocrine-immune (NEI) network. Methods The osteoporosis model was established by ovariectomy and randomly divided into model control group, Estradiol valerate group, Guben Zenggu Decoction high, medium and low concentration group. Selected the same age rats as the blank control group, after intervention, 10 rats were randomly selected at the time of 4th, 8th, 12th and 16th weeks from each group, After the success of the model and the successful intervention of Guben Zenggu Decoction for 4, 8, 12 and 16 weeks, the blood were taken from the heart, the serum bone alkaline phosphatase (BALP) levels was measured by the enzyme-linked immunosorbent assay (ELISA), and the thalamus, pituitary, spleen, adrenal, femoral tissue of rats were collected to detected the expressions of CaM and CaMKⅡ mRNA by using real-time fluorescence quantitative polymerase chain reaction (RT-PCR). Results At different time points (4 weeks, 8 CM（155mmweeks, 12 weeks, 16 weeks), the expressions of serum BALP and CaM and CaMKⅡ mRNA in NEI network in model group were significantly different from those in blank control group ( PCaM mRNA expression in hypothalamus and pituitary tissue were significantly increased in estradiol valerate and Guben Zenggu Decoction groups ( PCaM mRNA in adrenal gland, spleen and femur and the expression of CaMKⅡ mRNA in hypothalamus, pituitary, adrenal gland, spleen and femur were significantly decreased ( PCaM mRNA and CaMKⅡ mRNA in femoral and adrenal tissues was significantly different ( PCaM mRNA and CaMKⅡ mRNA in hypothalamus, pituitary, adrenal gland, spleen and femur, thus play a role in prevention and treatment of osteoporosis by regulating NEI network.