背根节(dorsal root ganglion,DRG)损伤或炎症可导致DRG神经元兴奋性异常增强和痛觉过敏。我们近期研究显示,长期慢性在体压迫(chronic compression of DRG,CCD)或急性离体分离(acute dissociation of DRG,ADD)背根节导致的神经元兴奋...背根节(dorsal root ganglion,DRG)损伤或炎症可导致DRG神经元兴奋性异常增强和痛觉过敏。我们近期研究显示,长期慢性在体压迫(chronic compression of DRG,CCD)或急性离体分离(acute dissociation of DRG,ADD)背根节导致的神经元兴奋性异常增强和痛觉过敏受环鸟苷酸(cGMP)-蛋白激酶G(PKG)信号通路活动的调控。本研究采用大鼠CCD模型和ADD模型,直接在DRG上检测cGMP浓度和PKG mRNA及其蛋白质的表达,进一步证明了cGMP-PKG信号通路活动在CCD和ADD DRG所致神经元兴奋性异常增强和痛觉过敏中的重要作用。酶联免疫吸附测定(ELISA)和逆转录聚合酶链反应(RT-PCR)的实验结果显示,CCD或ADD明显增高DRG内的cGMP浓度,上调I型PKG mRNA和PKG蛋白质表达。电生理膜片钳全细胞记录结果显示,CCD和ADD显著增强伤害特异性DRG细胞的兴奋性及其对cGMP-PKG信号通路激动剂的反应强度。增强的细胞兴奋性可以被cGMP-PKG通路阻断剂所抑制。在体压迫DRG的椎间孔内注射cGMP-PKG抑制剂显著减轻痛觉过敏。以上研究结果表明,CCD和ADD可以激活DRG细胞内的cGMP-PKG信号通路,而损伤的DRG细胞的超兴奋性和痛觉过敏的维持则需要cGMP-PKG信号通路处于持续的激活状态。展开更多
The activation of the Wnt/β-catenin signaling cascade has been well studied and documented in colorectal cancer（CRC）.The long-term use of non-steroidal anti-inflammatory drugs（NSAIDs） has been shown to reduce the...The activation of the Wnt/β-catenin signaling cascade has been well studied and documented in colorectal cancer（CRC）.The long-term use of non-steroidal anti-inflammatory drugs（NSAIDs） has been shown to reduce the incidence and risk of death from CRC in numerous epidemiological studies.The NSA1 D sulindac has also been reported to cause regression of precancerous adenomas in individuals with familial adenomatous polyposis who are at high risk of developing CRC.The mechanism responsible for cancer chemopreventive activity of NSAIDs is not well understood but may be unrelated to their cyclooxygenase inhibitory activity.Emerging evidence suggests that sulindac inhibits the growth of colon tumor cells by suppressing the activity of certain phosphodiesterase isozymes to activate cGMβ-dependent protein kinase,PKG,through the elevation of the second messenger cyclic guanosine monophosphote,cGMP.PKG activation has been shown to inhibit the nuclear translocation of β-catenin,reduce β-catenin mRNA and protein levels,and suppress the transcriptional activity of β-catenin.This review describes the relationship between the Wnt/β-catenin signaling cascade and the activation of PKG through PDE inhibition and elevation of intracellular cGMP levels.展开更多
目的:探讨藏药三味檀香散(Tsantan Sumtang of Tibetan medicine,TS)舒张肺小动脉的物质基础和作用机制。方法:使用中医药整合药理学研究平台(Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine,TCMIP...目的:探讨藏药三味檀香散(Tsantan Sumtang of Tibetan medicine,TS)舒张肺小动脉的物质基础和作用机制。方法:使用中医药整合药理学研究平台(Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine,TCMIP)分析方剂中化学成分及疾病靶点,构建“方剂-疾病-靶点”的相互作用网络和“方剂-中药材-成分-疾病-靶点-通路”的多维网络图,利用分子对接进一步验证TS有效成分与关键靶点的互作关系。利用离体血管环实验评价TS有效成分对肺血管的舒张作用。通过构建低氧诱导的肺动脉平滑肌细胞(pulmonary artery smooth muscle cells,PASMCs)增殖模型,利用激光共聚焦检测细胞内Ca^(2+)浓度,ELISA法用于分析TS有效成分对环化核苷酸(cyclic guanosine monophosphate,cGMP)和环腺苷酸(cyclic adenosine monophosphate,cAMP)含量的影响。结果:“方剂-疾病-靶点”的相互作用网络和“方剂-中药材-成分-疾病-靶点-通路”的多维网络图结果显示,TS包含的3味药材中有19个活性成分作用于136个靶点参与22条信号通路的调控,与治疗高原性肺动脉高压密切相关。分子对接结果显示TS有效成分与关键靶点具有较强的结合力,结合能前3位的成分为柚皮素、鞣花酸、西托糖苷,用这3个单体进行离体血管环实验,结果显示柚皮素、鞣花酸、西托糖苷对去甲肾上腺素(norephinephrine,NE)预收缩的大鼠肺小动脉血管环具有舒张作用,舒张率分别为(71.55±3.17)%,(68.91±17.72)%和(75.08±4.35)%。经一氧化氮合酶(nitric oxide synthase,NOS)抑制剂N-硝基-L-精氨酸甲酯盐酸盐(N-nitro-L-arginine methyl ester hydrochloride,L-NAME)和环氧合酶(cyclooxygenase,COX)抑制剂吲哚美辛(indomethacin,INDO)预处理后,柚皮素、鞣花酸和西托糖苷对NE预收缩的血管舒张作用明显减弱;格列苯脲的预处理降低了柚皮素、鞣花酸和西托糖苷的舒血管效应,K_(V)通道抑制剂4-AP干预后,降低了柚皮素和西托糖苷对NE预收缩的血管舒张作用;K_(IR)通道抑制剂BaCl_(2)的预处理显著降低了鞣花酸、西托糖苷的舒血管效应;四乙基氯化铵的干预仅降低了西托糖苷的舒血管效应。细胞实验结果显示西托糖苷能够显著降低低氧诱导的PASMCs中Ca^(2+)浓度,显著增加cGMP和cAMP的含量。结论:TS能够通过调控cGMP-PKG信号通路,激活PASMCs中的Kv通道抑制低氧诱导的PASMCs增殖舒张血管。展开更多
文摘The activation of the Wnt/β-catenin signaling cascade has been well studied and documented in colorectal cancer（CRC）.The long-term use of non-steroidal anti-inflammatory drugs（NSAIDs） has been shown to reduce the incidence and risk of death from CRC in numerous epidemiological studies.The NSA1 D sulindac has also been reported to cause regression of precancerous adenomas in individuals with familial adenomatous polyposis who are at high risk of developing CRC.The mechanism responsible for cancer chemopreventive activity of NSAIDs is not well understood but may be unrelated to their cyclooxygenase inhibitory activity.Emerging evidence suggests that sulindac inhibits the growth of colon tumor cells by suppressing the activity of certain phosphodiesterase isozymes to activate cGMβ-dependent protein kinase,PKG,through the elevation of the second messenger cyclic guanosine monophosphote,cGMP.PKG activation has been shown to inhibit the nuclear translocation of β-catenin,reduce β-catenin mRNA and protein levels,and suppress the transcriptional activity of β-catenin.This review describes the relationship between the Wnt/β-catenin signaling cascade and the activation of PKG through PDE inhibition and elevation of intracellular cGMP levels.
文摘目的:探讨藏药三味檀香散(Tsantan Sumtang of Tibetan medicine,TS)舒张肺小动脉的物质基础和作用机制。方法:使用中医药整合药理学研究平台(Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine,TCMIP)分析方剂中化学成分及疾病靶点,构建“方剂-疾病-靶点”的相互作用网络和“方剂-中药材-成分-疾病-靶点-通路”的多维网络图,利用分子对接进一步验证TS有效成分与关键靶点的互作关系。利用离体血管环实验评价TS有效成分对肺血管的舒张作用。通过构建低氧诱导的肺动脉平滑肌细胞(pulmonary artery smooth muscle cells,PASMCs)增殖模型,利用激光共聚焦检测细胞内Ca^(2+)浓度,ELISA法用于分析TS有效成分对环化核苷酸(cyclic guanosine monophosphate,cGMP)和环腺苷酸(cyclic adenosine monophosphate,cAMP)含量的影响。结果:“方剂-疾病-靶点”的相互作用网络和“方剂-中药材-成分-疾病-靶点-通路”的多维网络图结果显示,TS包含的3味药材中有19个活性成分作用于136个靶点参与22条信号通路的调控,与治疗高原性肺动脉高压密切相关。分子对接结果显示TS有效成分与关键靶点具有较强的结合力,结合能前3位的成分为柚皮素、鞣花酸、西托糖苷,用这3个单体进行离体血管环实验,结果显示柚皮素、鞣花酸、西托糖苷对去甲肾上腺素(norephinephrine,NE)预收缩的大鼠肺小动脉血管环具有舒张作用,舒张率分别为(71.55±3.17)%,(68.91±17.72)%和(75.08±4.35)%。经一氧化氮合酶(nitric oxide synthase,NOS)抑制剂N-硝基-L-精氨酸甲酯盐酸盐(N-nitro-L-arginine methyl ester hydrochloride,L-NAME)和环氧合酶(cyclooxygenase,COX)抑制剂吲哚美辛(indomethacin,INDO)预处理后,柚皮素、鞣花酸和西托糖苷对NE预收缩的血管舒张作用明显减弱;格列苯脲的预处理降低了柚皮素、鞣花酸和西托糖苷的舒血管效应,K_(V)通道抑制剂4-AP干预后,降低了柚皮素和西托糖苷对NE预收缩的血管舒张作用;K_(IR)通道抑制剂BaCl_(2)的预处理显著降低了鞣花酸、西托糖苷的舒血管效应;四乙基氯化铵的干预仅降低了西托糖苷的舒血管效应。细胞实验结果显示西托糖苷能够显著降低低氧诱导的PASMCs中Ca^(2+)浓度,显著增加cGMP和cAMP的含量。结论:TS能够通过调控cGMP-PKG信号通路,激活PASMCs中的Kv通道抑制低氧诱导的PASMCs增殖舒张血管。
