【摘要】 目的:鉴定转染大鼠组胺H3受体基因的HEK293细胞株受体表达情况,筛选具有H3受体拮抗活性的新型非咪唑类化合物。方法:利用RT-PCR和免疫印迹的方法检测转染细胞株组胺H3受体的表达,以阳性刺激物forskolin诱导细胞内cAMP含量升高为指标,初步筛选具有拮抗作用的化合物。结果:细胞鉴定结果显示,稳定转染H3受体基因的HEK293细胞株高表达大鼠H3受体。H3受体选择性激动剂(R)-α-甲基组胺可浓度依赖性抑制阳性刺激物forskolin(10μmol/L)诱导的cAMP含量升高(P<0.05),当(R)-α-甲基组胺浓度达到30 nmol/L时,抑制率接近平台值(P>0.05)。而H3受体拮抗剂噻普酰胺和新合成的非咪唑类化合物XHA23及XHA25则能够浓度依赖性阻断(R)-α-甲基组胺对forskolin诱导的cAMP含量升高的抑制作用(P<0.05),IC50分别为3.62μmol/L、0.49μmol/L、0.14μmol/L。结论:高表达H3受体的HEK293细胞株可用于有H3受体拮抗活性化合物的筛选,而非咪唑类化合物XHA23和XHA25具有一定的H3受体拮抗剂样活性。更多还原

【Abstract】 Objective: To identify a HEK293 cell line containing stably-transfected H3R gene,and to screen the novel non-imidazole compounds with H3R antagonist activity.Methods: The expression of rat H3 receptor in cell line was detected by RT-PCR and Western blot.An elevation of intercellular cAMP concentration induced by forskolin was measured as the index for screening compounds with H3R antagonist activity.Results: The H3R-transfected HEK-293 cells stably expressed high level of rat H3 receptor mRNA and protein.Forskolin significantly increased intercellular cAMP concentration in the H3R-transfected HEK-293 cells.H3R agonist(R)-α-methyhistamine inhibited the forskolin-induced production of intercellular cAMP.H3R antagonist thioperamide and newly synthesized non-imidazole compounds XHA23 and XHA25 blocked(R)-α-methyhistamine reversal of forskolin-induced cAMP formation in a concentration-dependent manner,and the IC50 values were 3.62 μmol/L,0.49 μmol/L,0.14 μmol/L,respectively.Conclusions: The H3R-transfected HEK293 cells stably express high level of rat H3 receptor,and can be used for screening compounds with H3R antagonist activity.The non-imidazole compounds XHA23 and XHA25 may have H3R antagonist activity.更多还原