Modulation Effect Of LW-AFC On The Unbalance Of Neuroimmunomodulation Network Induced By Stress

Objective: Mice models of three types of stress including psychological stress, physical stress and integrated stress were established to simulate the virous stress condition faced by soldiers, and then the imbalance characteristics of neuroimmunomodulation network (NIM) induced by these stress and the modulation effect of LW-AFC on it were studied.Methods: Psychological stress was established by tail suspension, physical stress by forced swimming, and integrated stress by unpredictable programmed stimuli. Drug or vehicle was given by oral administration. After the experiment blood sample was taken through the ophthalmic venous plexus with micro-hematocrit tubes and serum was extracted by centrifugation and then stored at ?20℃until hormone determination. The adrenal gland, thymus, spleen, testicular or ovaries was isolated and weighted. The level of IgG in the suspension of cultured spleen cells, VitC level in the homogenated suspension of adrenal gland and the serum corticosterone was measured by ELISA method. Level of serum estradiol, CRH and GnRH in hypothalamus and ACTH in pituitary was determined by radioimmunoassay and serum testosterone level by immunochemiluminescence. Protein expression of LH in pituitary was measured by western blotting. Proliferation and subcyte of T lymphocyte was determined by 3H-TdR incorporation and FACS respectively.Results: (1) In tail-suspension mice, serum CORT level and ACTH in pituitary was significantly increased, while CRH in hypothalamus was decreased, indicating the disorder of HPA axis. The stress-loaded mice also showed dysfunction of HPO axis as the diestrus was significantly prolonged; serum E2 during diestrus was increased, while pituitary LH expression and hypothalamus GnRH level was markedly decreased. Besides, the weight and IgG secretion of spleen decreased, indicating the stressed mice also showed immunosuppression. These results indicate the disfunction of neuroendocrine and immune system in psychological stress mice established by tail suspension.Oral administration of LW-AFC could anesis or reverses the change of hypothalamus CRH, pituitary ACTH and serum CORT in model mice to some extent. It showed significant modulation effect in increasing GnRH and LH secretion, prolonging estruation and decreasing serum E2 level. Detection of immunofunction showed the IgG secretion of spleen cell could also be increased by LW-AFC administration. These results indicate that LW-AFC can regulate the unbalance of NIM network in psychological stress mice.(2) In forced swimming mice, moving time and distance was significantly decreased showed by open field test, but mistaken frequency and latency did not change much showed by jumping stand test, indicating fatigue stress can induce depressive state without changing the passive avoidance response. The HPA axis in fatigue stress mice was over activated, as the adrenal gland showed hypertrophy and structural abnormalities, the CORT precursor, VitC, in adrenal markedly decreased, serum CORT significantly increased, and the upper hormone, ACTH in pituitary and CRH in hypothalamus decreased. Meanwhile, serum T and hypothalamus GnRH level was decreased, according with the weight loss and structural abnormalities of testis, indicating the function of HPG axis was down regulated. At the same time, all the immune indexes were down regulated including IgG level in suspension of cultured spleen cells, proliferation of spleen cells stimulated by ConA and LPS, and the ratio of Th and Ts cells in spleen, showing the lower function of immune system. These results indicate that physical stress loaded by forced swimming can induce the over activation of HPA axis and down-regulation of HPG axis and immune function.Oral administration of LW-AFC could significantly increase the moving time and distance, reverse the change of hypothalamus CRH, pituitary ACTH and adrenal VitC level. It also showed modulation effect in decreasing serum CORT, increasing hypothalamus GnRH and serum T level, and restoring the abnormal structure of testis. Meanwhile, the ratio of Th and Ts cells in spleen, proliferation of spleen cells stimulated by ConA and LPS and IgG secretion of spleen cell was upregulated. These results indicate that LW-AFC could anesis the depressive state, regulate the unbalance of HPA axis and HPG axis and upregulate the function of immune system. (3) In unpredictable programmed stimuli stressed mice, the adrenal gland became hypertrophy. At the same time the serum CORT level was significantly increased, while VitC in adrenal, ACTH in pituitary and CRH in hypothalamus markedly decreased, indicating the disorder of HPA axis. Serum T in model mice was also increased, but GnRH in hypothalamus was decreased, according with the weight loss of testis. Besides, all the immune indexes were down regulated, including the proliferation of spleen cells stimulated by ConA and LPS, IgG level in suspension of cultured spleen cells, and the ratio of Th and Ts cells in spleen, according with the weight loss of spleen showing the lower function of immune system. These results indicate that integrated stress can induce the imbalance of HPA axis and HPG axis and down-regulation of immune function.Oral administration of LW-AFC could improve or reverse the change of hypothalamus CRH, pituitary ACTH, serum CORT and adrenal VitC in model mice significantly, and suppressed the adrenal gland hypertrophy. At the same time, it could reverse serum T, hypothalamus GnRH level, and increase testis weight to modulate the disorder of HPG axis. Besides, results of the immunofunction showed that the proliferation of spleen cells stimulated by ConA and LPS, the IgG secretion of spleen cell could also be increased by LW-AFC administration. These results indicate that LW-AFC can regulate the imbalance of HPA and HPG axis and also improve the function of immune system.Conclusions: All of the three types of stress, psychological, physical and integrated stress, can induce the dysfunction of HPA axis, HPG axis and the immune system, indicating the unbalance of NIM network representing by dysfunction of neuroendocrine system and immune system was the common pathophysiology of the various kinds of stress. Yet different kind of stress might induce different characteristic of the unbalance. LW-AFC can modulate the disorder of NIM network through ameliorating or reversing the dysfunction of the HPA axis, HPG axis and the immune system, suggesting that LW-AFC might possibly be developed as a Traditional Chinese Medicine in anti-stress.