Qiao Wei-Li, Yang Wen-Xue, Liu Lei, Shi Yue, Cui Jie, Liu Hong, Yan Chang-Dong
Department of Physiology; Institute of Cardiovascular Disease; Department of Pathology; Jiangsu Province Key Laboratory of Anesthesiology, Xuzhou Medical College, Xuzhou 221004, China.
Sheng Li Xue Bao. 2014 Jun 25;66(3):276-82.
The present study was aimed to observe the protective effect of exogenous hydrogen sulfide (H₂S) on vascular structural and functional changes of aorta in D-galactose-induced subacute aging rats. Adult male SD rats were randomly divided to five groups: the vehicle group, the D-galactose (D-gal) group, and the three NaHS groups treated with low (1 μmol·kg⁻¹·d⁻¹), middle (10 μmol·kg⁻¹·d⁻¹) or high (100 μmol·kg⁻¹·d⁻¹) dose of NaHS respectively. The D-gal group rats were given subcutaneously injection of 125 mg/kg D-gal per day for eight weeks to induce subacute aging model. In the NaHS group, D-gal was administered as above but with NaHS intraperitoneally injected at a dosage of 1, 10, 100 μmol·kg⁻¹·d⁻¹ respectively. Equivalent volumes of saline were administered per day for eight weeks in vehicle group. Morphological changes of aorta were observed by HE and Masson staining. The level of H₂S in serum, the activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA), as well as anti-superoxide anions in vascular tissue were determined by spectrophotometry. Angiotensin II (AngII) levels in plasma were measured using competitive enzyme immunoassay. The expression of angiotensin II type 1 receptor (AT1R) in aorta was determined by Western blot. The results showed that the aging aortic morphologic changes in model rats were ameliorated in NaHS groups. Decreased vascular endothelial exfoliative cells and vascular smooth muscle cell (SMC) proliferation were shown in NaHS groups by HE staining. Masson staining analysis showed reduced relative contents of collagen fibers (P < 0.05) and SMC (P < 0.05) in NaHS groups. Compared to vehicle group, serum concentration of H₂S in D-gal group was decreased, while it was increased in NaHS groups after treatment with NaHS (P < 0.05). In the D-gal group, the concentration of AngII in plasma was significantly increased compared with that in vehicle group, while it was decreased in NaHS groups (P < 0.05). Moreover, levels of vascular tissue anti-superoxide anion and the activity of SOD were obviously higher, MDA was significantly lower in all NaHS treated groups than those in the D-gal group respectively (P < 0.05). Western blot analysis showed that the expression of AT1R was increased in D-gal group compared with that in vehicle group, while it was decreased after treatment with NaHS compared with that in D-gal group (P < 0.05). These results suggest that exogenous H₂S can ameliorate the age-related changes of aortic morphology, decrease the concentration of AngII in plasma, down-regulate the expression of AT1R in vascular tissue, and mitigate the level of oxidative stress. These changes delay the vascular aging in aging rats ultimately.
本研究旨在观察外源性硫化氢(H₂S)对D-半乳糖诱导的亚急性衰老大鼠主动脉血管结构和功能变化的保护作用。成年雄性SD大鼠随机分为五组:溶剂对照组、D-半乳糖(D-gal)组、低剂量(1 μmol·kg⁻¹·d⁻¹)、中剂量(10 μmol·kg⁻¹·d⁻¹)和高剂量(100 μmol·kg⁻¹·d⁻¹)NaHS处理组。D-gal组大鼠每天皮下注射125 mg/kg D-半乳糖,持续8周以诱导亚急性衰老模型。在NaHS组中,按上述方法给予D-半乳糖,但分别腹腔注射剂量为1、10、100 μmol·kg⁻¹·d⁻¹的NaHS。溶剂对照组每天给予等量体积的生理盐水,持续8周。通过HE和Masson染色观察主动脉的形态学变化。采用分光光度法测定血清中H₂S水平、超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量,以及血管组织中的抗超氧阴离子。采用竞争性酶免疫测定法测定血浆中血管紧张素II(AngII)水平。通过蛋白质免疫印迹法测定主动脉中血管紧张素II 1型受体(AT1R)的表达。结果显示,NaHS组改善了模型大鼠衰老主动脉的形态学变化。HE染色显示,NaHS组血管内皮剥脱细胞减少,血管平滑肌细胞(SMC)增殖减少。Masson染色分析显示,NaHS组胶原纤维(P < 0.05)和SMC(P < 0.05)的相对含量降低。与溶剂对照组相比,D-gal组血清H₂S浓度降低,而NaHS处理后NaHS组血清H₂S浓度升高(P < 0.05)。与溶剂对照组相比,D-gal组血浆中AngII浓度显著升高,而NaHS组血浆中AngII浓度降低(P < 0.05)。此外,所有NaHS处理组血管组织抗超氧阴离子水平和SOD活性均明显高于D-gal组,MDA含量明显低于D-gal组(P < 0.05)。蛋白质免疫印迹分析显示,与溶剂对照组相比,D-gal组AT1R表达增加,而与D-gal组相比,NaHS处理后AT1R表达降低(P < 0.05)。这些结果表明,外源性H₂S可改善主动脉形态的年龄相关变化,降低血浆中AngII浓度,下调血管组织中AT1R的表达,并减轻氧化应激水平。这些变化最终延缓了衰老大鼠的血管衰老。