| Objective1. To study the effects on the mRNA expression of 11 kinds of neuroendocrine hormone, peptide and receptor in hypothalamus and the relationship between the effect and time following acupuncture.2. To analyze the expression coincidence of acupuncture in the mRNA level and protein level by semi-quantitative the nuropeptide produce in different time after acupuncture through the RT-PCR and immunohistochemistry technique.3. To observe the effect of acupuncture on the expression of signal transduction and the relationship between effect and time following acupuncture for studying the protection mechanisms of acupuncture.4. To explore the modulation of acupuncture ST36 for the protection effect in stress ulcer animal model.Methods1. RT-PCR technique was used to observe the influence of acupuncture ST36 on the expression of OT, AVP, SP, POMC, IL-1beta,GnRH, Era, ERP,AR,TRH and CRH in hypothalamus and the relationship between effect and time underlying acupuncturing.2. Southern blot hybridization and immunohistochemistry technique were used to study the expression in mRNA level and protein level of neuropeptides (OT, AVP, SP and POMC) in hypothalamus stimulated by acupuncture ST36.3. Western blot hybridization was used to study the P38MAPK expressionof hypothalamus in protein level in different time post-acupuncture.4. RT-PCR technique, ulcer index estimate , HE -stain and radioimmunoassay were used to study the effect of acupuncture on mRNA expression in hypothalamus of NOS , ET-1, cortisol concentration in serum and stomach morphology in rats with stress ulcer.Results1. The result of RT-PCR for 11 kinds of neuroendocrine hormone, peptide and receptor in hypothalamus after acupuncture(1) Effect on OT mRNA expression: OT mRNA expression was increased significantly in para-acupoint group in 2-4hours post-acupuncturing ST36; and restored hi 6-8 hours post-acupuncture.(2) Effect on AVP mRNA expression: AVP mRNA expression was inhibition in acupoint group in 2-8 hours post-acupuncture.(3 ) Effect on POMC mRNA expression: POMC mRNA expression was suppression in 2 hours post-acupuncture, began to increase in 4 hours post-acupuncture, remained increase at 6 hours post-acupuncture , and restore in 8 hours post-acupuncture.(4 ) Effect on SP mRNA expression: SP mRNA expression was increased in acupoint group in 2-4 hours post-acupuncture. Adversely, the expression in para-acupoint group began to increase in 6-8 hours post-acupuncture(5) Effect on Era: Era mRNA expression in acupoint group began to increase at 2 hours post-acupoit, and remained at 4 hours post-acupuncture, and rehabilitation in 6-8 hours post-acupuncture.(6) GnRH mRNA expression was inhibited at 4 hours post-acupuncture inacupiont group, began to increase at 6 hours post-acupuncture in acupoint group; The effect was disappeared at 6 hourspost-acupuncture.2. The result of semi-quantitative estimate for OT, AVP, SP and POMC were coincidence with the RT-PCR results, Verified the correctness of RT-PCR results. Expression results in protein level: (1) OT: Expression in para-acupoint was increased and distribution was extensive relatively involved not only large cell neuron but also mini neuron as while as the hormone excretion in brain tissue . (2 ) AVP: Expression in para-acpoint was significantly decreased, and expression was mainly located in cytoplasm mini cell, a few in large cell cytoplasm. (3 ) SP: Expression in acupoint group was significantly increased, expression in para-acupoint group was slightly increased alsoo Expression site was in cytoplasm of large cell. (4) beta-EP: Expression in acupoint group was increased, located mainly in large cell cytoplasm.3. Acupuncturing ST36 inhibited P38MAPK expression in hypothalamus, and remained in 4 hours post-acupuncture.4. (1) The Ulcer Index was significant decreased in stress-induced ulcer ratsby acupuncturing contrast with only stress-induced ulcer rats. (2) Acupuncture inhibited the cortisol concentration serum in rat...
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