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目的:研究气体信号分子一氧化氮(NO)、一氧化碳(CO)在大鼠肝硬化中对硫化氢(H2S)/胱硫酶-γ-裂解酶(CSE)体系的影响。方法:将SD大鼠随机分为3组:正常对照组(NC组)、肝硬化模型组(CIRM组)、肝硬化模型+左旋硝基精氨酸甲酯(L-NAME)+锌原卟啉-Ⅸ(Znpp-Ⅸ)组(CIRM+L-NAME+ZnPP-Ⅸ组)。采用复合因素法建立大鼠肝硬化模型。运用敏感硫电极方法检测大鼠血浆中H2S的含量;应用免疫组织化学方法对大鼠门静脉(PV)上CSE蛋白进行定位及半定量分析;利用蛋白质免疫印迹技术(Western blot)测定大鼠肝组织中CSE蛋白的表达量。结果:在大鼠肝硬化形成后,血浆H2S含量显著降低,CIRM组〔(134.49±12.25)umol/L〕与NC组〔(180.34±11.71)umol/L〕比较有显著性差异(P<0.01);门静脉上CSE蛋白的表达〔NC组、CIRM组分别为(139.19±1.13)、(153.68±0.90)〕及其在肝组织中的蛋白含量〔NC组、CIRM组分别为(159.30±1.37)、(121.72±1.61)〕显著降低(P<0.01);当外源性给予L-NAME、ZnPP-Ⅸ后,CIRM+L-NAME+ZnPP-Ⅸ组血浆H2S含量〔(160.81±6.79)u-mol/L〕显著高于CIRM组〔(134.49±12.25)umol/L,P<0.01〕,同时,CIRM+L-NAME+ZnPP-Ⅸ组门静脉上CSE蛋白的表达(148.72±1.39)及其在肝组织中的蛋白含量(142.79±1.13)显著高于CIRM组(P<0.01)。结论:在大鼠肝硬化形成中,NO、CO可以上调H2S/CSE体系。
AIM: To investigate the effects of gas signaling molecules nitric oxide (NO) and carbon monoxide (CO) on hydrogen sulfide (H2S) / cystathionine-γ-lyase (CSE) system in rat liver cirrhosis. Methods: The SD rats were randomly divided into three groups: normal control group (NC group), cirrhosis model group (CIRM group), cirrhosis model + L-NAME + zinc protoporphyrin (Znpp-IX) group (CIRM + L-NAME + ZnPP-IX group). The compound factor method was used to establish the rat model of liver cirrhosis. The contents of H2S in rat plasma were detected by the sensitive sulfur electrode method. The CSE protein in the portal vein (PV) was localized and semi-quantitatively analyzed by immunohistochemical method. The rat liver tissue was determined by Western blot In CSE protein expression. Results: After the formation of cirrhosis in rats, the content of H2S in plasma decreased significantly (P <0.01, (134.49 ± 12.25) μmol / L in CIRM group compared with that in NC group (180.34 ± 11.71 umol / L〕 ). The expression of CSE protein in portal vein (NC group, CIRM group was (139.19 ± 1.13), (153.68 ± 0.90)] and its content in liver tissue [NC group, CIRM group were (159.30 ± 1.37) (121.72 ± 1.61)〕 (P <0.01). After exogenous administration of L-NAME and ZnPP-Ⅸ, the content of plasma H2S in CIRM + L-NAME + ZnPP-Ⅸ group was (160.81 ± 6.79) mol / L] was significantly higher than that of CIRM group [(134.49 ± 12.25) μmol / L, P <0.01〕. Meanwhile, the expression of CSE protein in the portal vein of CIRM + L-NAME + ZnPP-Ⅸ group was (148.72 ± 1.39) The protein content in liver tissue (142.79 ± 1.13) was significantly higher than that in CIRM group (P <0.01). Conclusion: NO and CO can up-regulate the H2S / CSE system in the formation of cirrhosis in rats.