腹膜淋巴孔与淋巴转归NO-cGMP-Ca^(2+)信号转导途径研究  

Effects of nitric oxide on peritoneal lymphatic stomata and lymph drainage via NO-cGMP-Ca^(2+) pathway

作　　者：李燕园[1] 李继承[1] 

机构地区：[1]浙江大学细胞生物学研究所淋巴学研究室,杭州310031

出　　处：《生理学报》2005年第1期45-53,共9页Acta Physiologica Sinica

基　　金：This work was supported by the National Natural Science Foundation of China (No. 30371815)

摘　　要：为研究一氧化氮(nitricoxide,NO)调节大鼠腹膜淋巴孔和淋巴吸收的细胞内信号转导机制,在体外培养的间皮细胞上,利用cGMP检测试剂盒和激光共聚焦扫描显微镜,研究NO对间皮细胞内cGMP和游离钙离子浓度([Ca2+]i)的影响;并利用动物实验研究NO-cGMP-Ca2+通路对大鼠腹膜淋巴孔和淋巴吸收的影响。结果发现:(1)与对照组相比,NO供体Sper/NO(spermine/nitricoxidecomplex)可以剂量依赖性地升高间皮细胞内cGMP的浓度(P<0.01)。此作用可被可溶性鸟苷酸环化酶(solubleguanylylcyclase,sGC)特异性抑制剂1H-[1,2,4]噁二唑[4,3-a]喹喔啉-1-one酮(1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one,ODQ)阻断(P<0.05,P<0.01)。Sper/NO可使间皮细胞内[Ca2+]i相对水平降低(P<0.05),但此效应可被ODQ阻断;L-型电压依赖性钙通道阻滞剂nifedipine,可使细胞内的[Ca2+]i在短时间内迅速明显下降(P<0.05),达平衡后再加入Sper/NO并不能引起[Ca2+]i的进一步改变(P>0.05);(2)NO可以剂量依赖性地提高大鼠膈组织淋巴孔最大分布面积(P<0.01)和对示踪剂的吸收量(P<0.05),ODQ可明显抑制NO引起的淋巴孔和淋巴吸收的改变(P<0.01)。该结果首次发现nifedipine能显著增加淋巴孔最大分布面积(P<0.01)及膈组织对台盼蓝的吸收量(P<0.05),而且nifedipine预?To study the cell signal transduction mechanism of nitric oxide (NO) on the peritoneal lymphatic stomata and lymph drainage in the rat, cGMP content were measured by a commercially available radioimmunoassay kit, and the [Ca2+]i were observed by a confocal laser scanning microscope in the cultured peritoneal mesothelial cell. Animal experiment was practiced to study the effect of NO-cGMP-Ca2+ pathway on the lymphatic stomata and lymph absorption. The results showed that: (1) Sper/NO increased cGMP of the rat peritoneal mesothelial cell (RPMC) in a dose-dependent manner (P<0.01) compared to the control group. This effect was blocked by 1H-[1,2,4] oxadiazolo [4,3-a] quinoxalin-1-one (ODQ) (P<0.05), a specific inhibitor of soluble guanylyl cyclase (sGC). The level of [Ca2+]i in single RPMC decreased by adding Sper/NO (P<0.05). Pretreatment with ODQ for 10 min blocked the Sper/NO-induced decrease in [Ca2+]i. L-typed calcium channel blocker nifedipine induced an immediate and marked decrease in [Ca2+]i(P<0.05). After [Ca2+]i reached a balance again, adding Sper/NO could not change [Ca2+]i (P>0.05). (2) Sper/NO increased the area of the stomata (P<0.01) and the quantity of the tracer in a dose-dependent manner (P<0.05) compared to the control group. Pretreatment with ODQ significantly inhibited Sper/NO-induced change of lymphatic stomata and lymph drainage (P<0.01). Nifedipine increased the opening area of the lymphatic stomata (P<0.01) and the concentration of absorbed trypan blue of the diaphragm (P<0.05). Sper/NO could not make a further change in the samples pretreated by nifedipine (P>0.05). The results indicate that NO can decrease [Ca2+]i in the RPMC through the NO-cGMP pathway. This procession is related with the L- type voltage-gated Ca2+ channel. NO enlarges the opening area of the lymphatic stomata and enhances the lymph drainage of tracer by NO-cGMP-[Ca2+]i pathway.

关 键 词：淋巴孔 信号转导 间皮细胞 一氧化氮 大鼠 

分 类 号：R33[医药卫生—人体生理学]