SNAP对大鼠生长激素腺瘤GH3细胞增殖、生长激素分泌的影响及机制探讨  

Effect and mechanism of SNAP on growth hormone secretion and cell proliferation in rat GH3 cells

作　　者：田春雷[1,2] 王雄伟[1,2] 王晓丹[1,2] 王旭光[1,2] 汪雷[1,2] 雷霆[3] 

机构地区：[1]宜昌市中心人民医院,湖北宜昌443003 [2]三峡大学神经病学研究所 [3]华中科技大学同济医学院附属同济医院

出　　处：《山东医药》2012年第2期15-17,共3页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(30672161)

摘　　要：目的观察一氧化氮(NO)释放剂S-亚硝基-N-乙酰青霉胺(SNAP)对大鼠生长激素(GH)腺瘤GH3细胞增殖、GH分泌的影响,并探讨其机制。方法将细胞分为A、B、C、D组,A组常规培养,B、C、D组分别加ghrelin、SNAP、SNAP+ghrelin培养。分别于培养第0、1、2、3、4、5、6天,用MTT法检测细胞吸光度值观察细胞增殖情况;培养2 h后,用ELISA法检测GH3细胞培养液中的GH,Western blot法检测GH3细胞中的磷酸化细胞外信号调节激酶(p-ERK)。结果 A组培养第2、3、4、5、6天时细胞吸光度值分别为0.381±0.006、1.664±0.005、3.411±0.005、5.221±0.005、10.055±0.005,B组分别为1.062±0.005、3.115±0.005、4.512±0.005、6.656±0.005、11.060±0.005,C组分别为0.161±0.006、0.413±0.005、0.931±0.005、1.861±0.005、5.612±0.005,D组分别为0.219±0.004、0.865±0.005、1.712±0.005、3.059±0.005、6.561±0.005;B组与A组、C组与A组、D组与B组比较,P均<0.01。培养2 h时,A、B、C、D组GH3细胞培养液的吸光度值分别为0.080±0.002、0.165±0.011、0.041±0.003、0.106±0.005,pERK表达量分别为0.301 8±0.004 5、0.682 5±0.003 8、0.192 3±0.008 6、0.298 5±0.006 7,B组与A组、C组与A组、D组与B组比较,P均<0.01。结论 SNAP可抑制ghrelin诱导的GH3细胞增殖及GH分泌,可能与其阻断ghrelin激活的ERK信号通路有关。Objective To obserbve the effects of S-nitroso-N-acetylpenicillamine （SNAP） on growth hormone （GH） secretion and cell proliferation in rat GH3 cells and explore the possible mechanism. Methods GH3 cells were divided into A, B, C, D group. GH3 cells in group A were cultured normally. GH3 cells in group B, C, D were cultured with ghrelin, SNAP, ghrelin ＋ SNAP. After 0,1,2,3 ,4,5 ,6 d, MTT was used to detected the absorbance value. After cultured for 2 h, ELISA was used to detected the GH in GH3 cells culture solution; Western blot was used to detected the phospho-excelluar signal-regulated kinase（p-ERK） in GH3 cells. Results The absorbanee value of GH3 cells cultured for 2,3,4,5,6 d in group A were 0. 381 ± 0. 006, 1. 664 ± 0. 005, 3.411 ± 0. 005, 5. 221 ± 0. 005, 10. 055 ± 0. 005 ; 1. 062 ± 0. 005, 3. 115±0.005, 4.512 ±0.005, 6.656 ±0.005,11.060±0.005 in group B; 0. 161 ±0.006, 0.413±0.005, 0.931 ± 0.005, 1.861 ±0.005, 5.612±0.005 in group C; 0.219 ±0.004, 0.865 ±0.005, 1.712 ±0.005, 3.059 ±0.005, 6. 561 ± 0. 005 in group D. There were significant difference between the absorbance value of GH3 cells in group B and A, CandA, DandB（allP〈0.01）. The GHlevelin group A,B,C,D cultured for2 h were0.080±0.002, 0.165 ± 0.011, 0.041 ±0.003, 0. 106 ±0.005; and the p-ERK were 0.301 8 ±0.004 5, 0.682 5 ±0.003 8, 0. 192 3 ± 0. 008 6, 0.298 5 ± 0. 006 7. There were significant difference between the GH level and p-ERK in GH3 cells in group B and A, C and A, D and B （all P 〈 0.01 ）. Conclusion SNAP reduces both basal and induced by ghrelin GH secretion and cell proliferation in rat GH3 cells via blocking ERK signaling pathway.

关 键 词：S-亚硝基-N-乙酰青霉胺 生长激素 细胞外信号调节激酶 

分 类 号：R739.41[医药卫生—肿瘤]