天麻素调节cAMP/PKA/GRK2信号通路对脑胶质瘤细胞增殖、凋亡的影响  

Impacts of Gastrodin on the Proliferation and Apoptosis of Glioma Cells by Regulating cAMP/PKA/GRK2 Signaling Pathway

作　　者：滑祥廷 洪文明[1] 张超勇[2] 王斌[1] Hua Xiangting;Hong Wenming;Zhang Chaoyong(Department of Neurosurgery,the First Affiliated Hospital of Anhui Medical University,Hefei 230022,China;Taihe County People’s Hospital,Fuyang 236600,China)

机构地区：[1]安徽医科大学第一附属医院神经外科,合肥230022 [2]安徽省太和县人民医院神经外科,阜阳236600

出　　处：《华中科技大学学报（医学版）》2022年第5期607-611,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.82003795);安徽省自然科学基金资助项目(No.1908085QH379)。

摘　　要：目的探讨天麻素(Gastrodin)对脑胶质瘤细胞增殖、凋亡的影响及作用机制。方法不同浓度的天麻素(2.5~80.0μmol/L)分别处理U87细胞24、48、72 h,检测细胞增殖确定最佳给药浓度;U87细胞分为对照组(Control组)、天麻素低、中、高浓度组(Gastrodin-L、Gastrodin-M、Gastrodin-H组,天麻素工作浓度分别为5、10、20μmol/L)、天麻素+cAMP/PKA信号通路抑制剂组(Gastrodin+H-89组,20μmol/L的天麻素与10μmol/L的H-89共同处理细胞),分别检测U87细胞增殖、凋亡及cAMP及Cyclin D1、p21、Bax、Bcl-2、Caspase-3、p-PKA、PKA、p-GRK2、GRK2的蛋白表达水平。结果2.5~80.0μmol/L的天麻素均可抑制U87细胞增殖,选择浓度为5、10、20μmol/L的天麻素进行后续实验。与Control组比较,Gastrodin-L、Gastrodin-M、Gastrodin-H组G_(0)/G_(1)期细胞比例、细胞凋亡率、cAMP、p21、Bax、Caspase-3、p-PKA、p-GRK2蛋白表达水平均显著升高(均P<0.05),S期与G_(2)/M期细胞比例、集落形成数、Cyclin D1、Bcl-2蛋白表达水平显著降低(均P<0.05),且呈药物浓度依赖性;与Gastrodin-H组比较,Gastrodin+H-89组G_(0)/G_(1)期细胞比例、细胞凋亡率、cAMP、p21、Bax、Caspase-3、p-PKA、p-GRK2蛋白表达水平显著降低(均P<0.05),S期与G_(2)/M期细胞比例、集落形成数、Cyclin D1、Bcl-2蛋白表达水平显著升高(均P<0.05)。结论天麻素可能通过激活cAMP/PKA/GRK2通路抑制脑胶质瘤细胞增殖,并诱导其凋亡。Objective To investigate the impacts of Gastrodin on the proliferation and apoptosis of glioma cells and its mechanism.Methods U87 cells were treated with different concentrations of Gastrodin(2.5-80.0μmol/L)for 24,48 and 72 h,respectively.The cell proliferation was detected to determine the optimal concentration of drug administration.U87 cells were divided into control group,gastrodin low,medium and high concentration groups(Gastrodin-L,Gastrodin-M,and Gastrodin-H groups,treated with gastrodin at 5,10,and 20μmol/L,respectively),Gastrodin+cAMP/PKA signaling pathway inhibitor group(Gastrodin+H-89 group,treated with 20μmol/L gastrodin and 10μmol/L H-89).U87 cell proliferation,apoptosis and levels of cAMP,Cyclin D1,p21,Bax,Bcl-2,Caspase-3,p-PKA,PKA,p-GRK2,GRK2 protein were detected respectively.Results Gastrodin at 2.5-80.0μmol/L could significantly inhibit the proliferation of U87 cells,and gastrodin concentrations of 5,10,and 20μmol/L were selected for subsequent experiments.Compared with control group,the G_(0)/G_(1) phase cell ratio,cell apoptosis rate,cAMP,p21,Bax,Caspase-3,p-PKA,p-GRK2 protein expression levels were obviously increased in the Gastrodin-L,Gastrodin-M,Gastrodin-H group(all P<0.05),the S phase and G_(2)/M phase ratios,the number of colonies,Cyclin D1 and Bcl-2 protein expression levels were obviously decreased(all P<0.05)in a drug concentration-dependent manner.Compared with the Gastrodin-H group,the G_(0)/G_(1) phase cell ratio,cell apoptosis rate,cAMP,p21,Bax,Caspase-3,p-PKA,p-GRK2 protein expression levels were obviously decreased in the Gastrodin+H-89 group(all P<0.05).The S phase and G_(2)/M phase ratios,the number of colonies,Cyclin D1 and Bcl-2 protein expression levels were obviously increased(all P<0.05).Conclusion Gastrodin inhibits the proliferation of glioma cells by activating the cAMP/PKA/GRK2 pathway,and induces apoptosis.

关 键 词：天麻素 环磷酸腺苷/蛋白激酶A/G蛋白偶联受体激酶2信号通路 胶质瘤 增殖 凋亡 

分 类 号：R739.41[医药卫生—肿瘤]