目的:探讨次乌头碱(HA)通过环磷酸鸟苷-腺苷酸合成酶(cGAS)/干扰素基因刺激因子(STING)信号通路对胃癌细胞增殖、迁移、侵袭和免疫逃逸的影响。方法:将BGC-823细胞分为对照组(NC组)、HA低剂量组(HA-L组,2μmol/L)、HA中剂量组(HA-M组,4...目的:探讨次乌头碱(HA)通过环磷酸鸟苷-腺苷酸合成酶(cGAS)/干扰素基因刺激因子(STING)信号通路对胃癌细胞增殖、迁移、侵袭和免疫逃逸的影响。方法:将BGC-823细胞分为对照组(NC组)、HA低剂量组(HA-L组,2μmol/L)、HA中剂量组(HA-M组,4μmol/L)、HA高剂量组(HA-H组,8μmol/L)、RU.521(cGAS抑制剂)组(1μmol/L)、HA-H+RU.521组(8μmol/L+1μmol/L),CCK-8、克隆形成实验检测细胞增殖;划痕愈合实验检测细胞迁移;Transwell实验检测细胞侵袭;ELISA检测细胞上清中趋化因子配体(CXCL)2、CXCL8水平;Western blot检测细胞中增殖细胞核抗原(PCNA)、基质金属蛋白酶(MMP)-9、cGAS、STING蛋白表达。将上述各组细胞分别与NK细胞共培养24 h,命名为NC共培养组、HA-L共培养组、HA-M共培养组、HA-H共培养组、RU.521共培养组、HA-H+RU.521共培养组,检测NK细胞杀伤力。结果:与NC组比较,HA-L组、HA-M组、HA-H组BGC-823细胞OD450(24 h)(0.87±0.08 vs 0.75±0.06、0.62±0.06、0.41±0.03)、克隆形成率[(47.75±2.13)%vs(41.12±1.81)%、(33.58±1.61)%、(19.95±0.84)%]、划痕愈合率[(43.37±2.08)%vs(36.65±1.54)%、(27.74±1.03)%、(13.36±0.62)%]、侵袭细胞数(78.85±3.67 vs 65.59±2.49、51.52±2.01、22.23±1.36)、CXCL2、CXCL8水平、PCNA、MMP-9蛋白表达降低,cGAS、STING蛋白表达升高(P<0.05);与NC组比较,RU.521组对应指标变化趋势与上述相反(P<0.05);与NC共培养组比较,HA-L共培养组、HA-M共培养组、HA-H共培养组NK细胞杀伤力增强,且呈剂量依赖性(P<0.05);与NC共培养组比较,RU.521共培养组对应指标变化趋势与上述相反(P<0.05);RU.521减弱了高剂量HA对BGC-823细胞增殖、迁移、侵袭、免疫逃逸的抑制作用。结论:HA可能通过激活cGAS-STING信号通路抑制胃癌细胞增殖、迁移、侵袭和免疫逃逸。展开更多
Abscisic acid (ABA) plays an important role in plant growth and developmental processes. Although some ABA signal molecules, such as cADPR, Ca2+, etc., have been reported, there. was no evidence proving the involvemen...Abscisic acid (ABA) plays an important role in plant growth and developmental processes. Although some ABA signal molecules, such as cADPR, Ca2+, etc., have been reported, there. was no evidence proving the involvement of cAMP in A-B-A, signal transduction. In this present study, the constructed gene ( rd29A-GUS) was transformed into Nicotiana tabacum, and calli was induced from the transgenic plant. The suspension cells obtained from the callus grew well and uniformly. Treatment of the suspension cells with ABA led to an increase in GUS activity, indicating that these transgenic suspension cells are useful for the study of ABA signaling. Addition of nicotinamide (cADPR inhibitor) or U-73122 (phospholiphase C inhibitor) could only partially inhibit the increase of GUS activity elicited by ABA. The inhibitory effect of nicotinamide was enhanced by application of K252a (inhibitor of protein kinase). Treatment of the suspension cells with 8-Br-cAMP, a membrane-permeable analogue of cAMP, could partially replace the effect of ABA. Furthermore, intracellular addition of IBMX (phosphodiesterase inhibitor) mimicked die effect of exogenous cAMP on the deduction of expression of rd29A promoter. These results suggested that cAMP was an important messenger in ABA signal transduction in tobacco suspension cell.展开更多
[目的]探究环磷酸鸟苷-腺苷酸合成酶/干扰素刺激基因(cGAS-STING)信号通路在急性心肌梗死(AMI)小鼠模型中表达及作用。[方法]选择10~12周龄C57野生型小鼠,分为假手术组(只穿线不结扎)、模型组(结扎冠脉左降前支建立AMI小鼠模型)、STING...[目的]探究环磷酸鸟苷-腺苷酸合成酶/干扰素刺激基因(cGAS-STING)信号通路在急性心肌梗死(AMI)小鼠模型中表达及作用。[方法]选择10~12周龄C57野生型小鼠,分为假手术组(只穿线不结扎)、模型组(结扎冠脉左降前支建立AMI小鼠模型)、STING抑制组(建模+腹腔注射5 mg/kg STING抑制剂C18)各10只,采用超声评估其心功能,HE染色、TUNEL法评估心肌组织病理情况及细胞凋亡,ELISA法检测血清炎性因子表达,Western Blot检测心肌组织cGAS、STING、IRF3及p-IRF3蛋白表达。[结果]与假手术组比较,模型组LVESD、LVEDD、心脏重量、左心室重量及其重量指数、心肌细胞凋亡率、血清TGF-β、IL-6、IL-18水平、心肌组织cGAS、STING、p-IRF3/IRF3蛋白表达明显升高,心脏梗死面积扩大,LVFS、LVEF下降;与模型组比较,STING抑制组LVEF、LVFS升高,LVESD、LVEDD、心脏重量、左心室重量及其重量指数、心肌细胞凋亡率、血清TGF-β、IL-6、IL-18水平、心肌组织cGAS(1.99±0.12 vs 0.73±0.09)、STING(1.41±0.18 vs 0.37±0.12)、p-IRF3/IRF3蛋白表达降低,梗死面积减少(P<0.05)。[结论]阻断cGAS-STING信号通路能改善AMI小鼠心功能,其机制可能与能减少心肌细胞凋亡和心室重构有关。展开更多
文摘目的:探讨次乌头碱(HA)通过环磷酸鸟苷-腺苷酸合成酶(cGAS)/干扰素基因刺激因子(STING)信号通路对胃癌细胞增殖、迁移、侵袭和免疫逃逸的影响。方法:将BGC-823细胞分为对照组(NC组)、HA低剂量组(HA-L组,2μmol/L)、HA中剂量组(HA-M组,4μmol/L)、HA高剂量组(HA-H组,8μmol/L)、RU.521(cGAS抑制剂)组(1μmol/L)、HA-H+RU.521组(8μmol/L+1μmol/L),CCK-8、克隆形成实验检测细胞增殖;划痕愈合实验检测细胞迁移;Transwell实验检测细胞侵袭;ELISA检测细胞上清中趋化因子配体(CXCL)2、CXCL8水平;Western blot检测细胞中增殖细胞核抗原(PCNA)、基质金属蛋白酶(MMP)-9、cGAS、STING蛋白表达。将上述各组细胞分别与NK细胞共培养24 h,命名为NC共培养组、HA-L共培养组、HA-M共培养组、HA-H共培养组、RU.521共培养组、HA-H+RU.521共培养组,检测NK细胞杀伤力。结果:与NC组比较,HA-L组、HA-M组、HA-H组BGC-823细胞OD450(24 h)(0.87±0.08 vs 0.75±0.06、0.62±0.06、0.41±0.03)、克隆形成率[(47.75±2.13)%vs(41.12±1.81)%、(33.58±1.61)%、(19.95±0.84)%]、划痕愈合率[(43.37±2.08)%vs(36.65±1.54)%、(27.74±1.03)%、(13.36±0.62)%]、侵袭细胞数(78.85±3.67 vs 65.59±2.49、51.52±2.01、22.23±1.36)、CXCL2、CXCL8水平、PCNA、MMP-9蛋白表达降低,cGAS、STING蛋白表达升高(P<0.05);与NC组比较,RU.521组对应指标变化趋势与上述相反(P<0.05);与NC共培养组比较,HA-L共培养组、HA-M共培养组、HA-H共培养组NK细胞杀伤力增强,且呈剂量依赖性(P<0.05);与NC共培养组比较,RU.521共培养组对应指标变化趋势与上述相反(P<0.05);RU.521减弱了高剂量HA对BGC-823细胞增殖、迁移、侵袭、免疫逃逸的抑制作用。结论:HA可能通过激活cGAS-STING信号通路抑制胃癌细胞增殖、迁移、侵袭和免疫逃逸。
文摘Abscisic acid (ABA) plays an important role in plant growth and developmental processes. Although some ABA signal molecules, such as cADPR, Ca2+, etc., have been reported, there. was no evidence proving the involvement of cAMP in A-B-A, signal transduction. In this present study, the constructed gene ( rd29A-GUS) was transformed into Nicotiana tabacum, and calli was induced from the transgenic plant. The suspension cells obtained from the callus grew well and uniformly. Treatment of the suspension cells with ABA led to an increase in GUS activity, indicating that these transgenic suspension cells are useful for the study of ABA signaling. Addition of nicotinamide (cADPR inhibitor) or U-73122 (phospholiphase C inhibitor) could only partially inhibit the increase of GUS activity elicited by ABA. The inhibitory effect of nicotinamide was enhanced by application of K252a (inhibitor of protein kinase). Treatment of the suspension cells with 8-Br-cAMP, a membrane-permeable analogue of cAMP, could partially replace the effect of ABA. Furthermore, intracellular addition of IBMX (phosphodiesterase inhibitor) mimicked die effect of exogenous cAMP on the deduction of expression of rd29A promoter. These results suggested that cAMP was an important messenger in ABA signal transduction in tobacco suspension cell.
文摘[目的]探究环磷酸鸟苷-腺苷酸合成酶/干扰素刺激基因(cGAS-STING)信号通路在急性心肌梗死(AMI)小鼠模型中表达及作用。[方法]选择10~12周龄C57野生型小鼠,分为假手术组(只穿线不结扎)、模型组(结扎冠脉左降前支建立AMI小鼠模型)、STING抑制组(建模+腹腔注射5 mg/kg STING抑制剂C18)各10只,采用超声评估其心功能,HE染色、TUNEL法评估心肌组织病理情况及细胞凋亡,ELISA法检测血清炎性因子表达,Western Blot检测心肌组织cGAS、STING、IRF3及p-IRF3蛋白表达。[结果]与假手术组比较,模型组LVESD、LVEDD、心脏重量、左心室重量及其重量指数、心肌细胞凋亡率、血清TGF-β、IL-6、IL-18水平、心肌组织cGAS、STING、p-IRF3/IRF3蛋白表达明显升高,心脏梗死面积扩大,LVFS、LVEF下降;与模型组比较,STING抑制组LVEF、LVFS升高,LVESD、LVEDD、心脏重量、左心室重量及其重量指数、心肌细胞凋亡率、血清TGF-β、IL-6、IL-18水平、心肌组织cGAS(1.99±0.12 vs 0.73±0.09)、STING(1.41±0.18 vs 0.37±0.12)、p-IRF3/IRF3蛋白表达降低,梗死面积减少(P<0.05)。[结论]阻断cGAS-STING信号通路能改善AMI小鼠心功能,其机制可能与能减少心肌细胞凋亡和心室重构有关。
