补肾方剂对GT1细胞株的作用及机理研究

To investigate the effects of drug sera from the animals fed "Kidney Tonifying" formula "Two Fairy Decoction" and its two divided "Kidney Warming" and "Yin Nourishing" combinations on GT1－7 cell line which was developed by targeting the SV40 T antigen to GnRH neuron in a transgenic mouse. Method: (1) GnRH secretion was measured by a newly radioimmunoassay to determine the effect of Kidney Tonifying formula "Two Fairy Decoction" and its two divided combinations "Kidney Warming" and "Yin Nourishing" on GT1－7 cell line. (2) Northern blot analysis of GnRH and IGF-I mRNA was performed. Result: (1) GnRH RIA developed by our laboratory was comparable to Knobil's laboratory. (2) " Two Fairy Decoction" and its two divided combinations all have a stimulating effect on GnRH secretion, the whole formula being better than each divided combinations of its ingredients. (3) Following an incubation for 24hr with 10% drug sera of " Two Fairy Decoction" and its one divided combination"Kidney Warming", the GnRH levels showed a little decline. While exposure to 10% drug sera of " Yin Nourishing " for 48hr increased GnRH mRNA levels to at least 1.5 - fold of the contrast, although without any detectable effect for the 24hr period. (4) the administration of "Two Fairy Decoction" and its two divided combinations for 24hr elevated IGF-I mRNA levels dramaticaly.(5)Some effective composition from " Two Fairy Decoction" have a stimulating effect on GnRH secretion.conclusion: Traditional Chinese medicine holds that the Kidney is essential in reproduction, which includes function of GnRH and its regulation of gonadotropin. The Kidney Tonifying Formula "Two Fairy Decoction" and its divided combinations are able to directly stimulate GnRH secretion and to regulate relative gene expression in vitro..

GT1细胞株是转入了SV40T抗原基因的GnRH神经内分泌细胞株，本课题旨在运用血清药理学的方法，结合放免技术、现代细胞生物学和分子生物学技术，研究中药补肾方剂及其拆方对该细胞株GnRH分泌和基因表达的作用及机理研究，为中医药治疗性腺轴衰老疾病和进一部探讨肾的本质提供实验依据，填补整体实验所无法解决的空白。