Objective To investigate the effect of H2S on lower limb ischemia-reperfusion （LIR） induced lung injury and explore the underlying mechanism. Methods Wistar rats were randomly divided into control group, IR group, I...Objective To investigate the effect of H2S on lower limb ischemia-reperfusion （LIR） induced lung injury and explore the underlying mechanism. Methods Wistar rats were randomly divided into control group, IR group, IR＋ Sodium Hydrosulphide （NariS） group and IR＋ DL-propargylglycine （PPG） group. IR group as lung injury model induced by LIR were given 4 h reperfusion following 4 h ischemia of bilateral hindlimbs with rubber bands. NariS （0.78 mg/kg） as exogenous H2S donor and PPG （60 mg/kg） which can suppress endogenous H2S production were administrated before LIR, respectively. The lungs were removed for histologic analysis, the determination of wet-to-dry weight ratios and the measurement of mRNA and protein levels of aquaporin-1 （AQP1）, aquaporin-5 （AQP5） as indexes of water transport abnormality, and mRNA and protein levels of Toll-like receptor 4 （TLR4）, myeloid differentiation primary-response gene 88 （MyD88） and p-NF-KB as indexes of inflammation. Results LIR induced lung injury was accompanied with upregulation of TLR4-Myd88-NF-κB pathway and downregulation of AQP1/AQP5. NariS pre-treatment reduced lung injury with increasing AQP1/AQP5 expression and inhibition of TLR4-Myd88-NF-KB pathway, but PPG adjusted AO.PJAO.Ps and TLR4 pathway to the opposite side and exacerbated lung injury. Conclusion Endogenous H2S, TLR4-Myd88-NF-κB pathway and AQP1/AQP5 were involved in LIR induced lung injury. Increased H2S would alleviate lung injury and the effect is at least partially depend on the adjustment of TLR4-Myd88-NF-κB pathway and AQP1/AQP5 expression to reduce inflammatory reaction and lessen pulmonary edema.展开更多
In this study, we amplified aqnaporin I（AQP1） promoter sequence with polymerase chain reaction（PCR）, then AQP1 promoter fragment and pGL2 basic vector were linked to create an artificial pGL2/AQP1 promoter re- por...In this study, we amplified aqnaporin I（AQP1） promoter sequence with polymerase chain reaction（PCR）, then AQP1 promoter fragment and pGL2 basic vector were linked to create an artificial pGL2/AQP1 promoter re- porter system. A certain concentration of 17β-estradiol（E2） activated pGL2/AQPlp, which demonstrated the pGL2/AQPlp transcriptional system effective. The pGL2/AQP1 promoter reporter system was applied to evaluate the activate effect on AQP1 of different kinds of phytoestrogens. Dual hiciferase reporter gene activity assay showed that a certain concentration phytoestrogens including daidzein and genistein can increase AQP1 promoter transcription activity. In addition, E2, daidzein and genistein can make AQP1 protein endogenous expression level increase and promote the function of water scretion. The result can guide the clinical application to treat the Sjogren＇s syndrome and provide a necessary molecular tool for the subsequent drug screening.展开更多
基金supported by the Military Health Care Foundation during the 12th Five-year Plan Period(11BZ21)the Military Scientific Research Foundation during the 12th Five-year Plan Period(BWS12J051)
文摘Objective To investigate the effect of H2S on lower limb ischemia-reperfusion （LIR） induced lung injury and explore the underlying mechanism. Methods Wistar rats were randomly divided into control group, IR group, IR＋ Sodium Hydrosulphide （NariS） group and IR＋ DL-propargylglycine （PPG） group. IR group as lung injury model induced by LIR were given 4 h reperfusion following 4 h ischemia of bilateral hindlimbs with rubber bands. NariS （0.78 mg/kg） as exogenous H2S donor and PPG （60 mg/kg） which can suppress endogenous H2S production were administrated before LIR, respectively. The lungs were removed for histologic analysis, the determination of wet-to-dry weight ratios and the measurement of mRNA and protein levels of aquaporin-1 （AQP1）, aquaporin-5 （AQP5） as indexes of water transport abnormality, and mRNA and protein levels of Toll-like receptor 4 （TLR4）, myeloid differentiation primary-response gene 88 （MyD88） and p-NF-KB as indexes of inflammation. Results LIR induced lung injury was accompanied with upregulation of TLR4-Myd88-NF-κB pathway and downregulation of AQP1/AQP5. NariS pre-treatment reduced lung injury with increasing AQP1/AQP5 expression and inhibition of TLR4-Myd88-NF-KB pathway, but PPG adjusted AO.PJAO.Ps and TLR4 pathway to the opposite side and exacerbated lung injury. Conclusion Endogenous H2S, TLR4-Myd88-NF-κB pathway and AQP1/AQP5 were involved in LIR induced lung injury. Increased H2S would alleviate lung injury and the effect is at least partially depend on the adjustment of TLR4-Myd88-NF-κB pathway and AQP1/AQP5 expression to reduce inflammatory reaction and lessen pulmonary edema.
文摘In this study, we amplified aqnaporin I（AQP1） promoter sequence with polymerase chain reaction（PCR）, then AQP1 promoter fragment and pGL2 basic vector were linked to create an artificial pGL2/AQP1 promoter re- porter system. A certain concentration of 17β-estradiol（E2） activated pGL2/AQPlp, which demonstrated the pGL2/AQPlp transcriptional system effective. The pGL2/AQP1 promoter reporter system was applied to evaluate the activate effect on AQP1 of different kinds of phytoestrogens. Dual hiciferase reporter gene activity assay showed that a certain concentration phytoestrogens including daidzein and genistein can increase AQP1 promoter transcription activity. In addition, E2, daidzein and genistein can make AQP1 protein endogenous expression level increase and promote the function of water scretion. The result can guide the clinical application to treat the Sjogren＇s syndrome and provide a necessary molecular tool for the subsequent drug screening.
