硫化氢对蝮蛇毒致大鼠急性肺损伤氧化应激的调节作用

发布时间：2019-04-20 19:39

【摘要】：目的：腹腔注射蝮蛇毒素建立急性肺损伤模型，观察增加或减少H_2S的含量对氧化保护及氧化损伤指标的影响，探讨H_2S/CSE体系对蝮蛇毒素致大鼠急性肺损伤氧化应激的调节作用。方法： 1.分组：采用完全随机设计将36只雄性清洁级SD大鼠分成6组，每组6只，分别为：①生理盐水对照组(N组)；②蝮蛇毒素组(S组)；③蝮蛇毒素＋低剂量NaHS组(S+L组)；④蝮蛇毒素＋中剂量NaHS组(S+M组)；⑤蝮蛇毒素＋高剂量NaHS组(S+H组)；⑥蝮蛇毒素＋PPG组(S+PPG组)。 2.方法：①将N组腹腔注射等容积生理盐水，其余组分别腹腔注射蝮蛇毒素。②3h后，N组和S组分别腹腔注射等容积生理盐水；S+L, S+M, S+H组和S+PPG组，分别腹腔注射0.78mg/kg,1.56mg/kg和3.12mg/kg剂量的NaHS和30mg/kg剂量的PPG。③6h后，将各组实验动物处死，采集心脏血并分离留取血清，取相应肺组织留作标本。④肺组织HE染色光镜观察病理学变化并行病理学评分，测定肺组织湿/干重比，去蛋白法检测血浆H_2S含量，亚甲基蓝法测定肺组织CSE活性，WST-1法测定肺组织匀浆超氧化物歧化酶(SOD)活力，酶促反应谷胱甘肽消耗法测定肺组织谷胱甘肽过氧化酶(GSH-Px)活力，酶联免疫法(ELISA)法测定肺组织8-异构前列腺素(8-iso-PGF2α)含量。结果： 1.肺组织湿/干重比(W/D)变化：与N组相比较，S组W/D值升高(P0.01)。与S组相比较，不认为S+L组W/D值下降存在差异(P0.05)；S+M, S+H组W/D值均有明显下降(P0.01)；S+PPG组W/D值升高(P0.05)。 2.光镜下肺组织病理学变化及肺损伤病理评分(IQA)：与N组正常肺组织相比，S组肺泡结构明显紊乱、破坏，肺泡结构不完整，明显肺泡水肿，大量透明膜形成，肺间质明显增厚，肺间质及肺泡大量红细胞及粒细胞浸润，可见肺不张，IQA显著升高(P0.01)。与S组相比较，S+L组肺组织损伤未见明显改善(P0.05)，S+M,S+H组上述肺损伤情况有所减轻，肺泡结构基本完整，但仍见肺泡间隔增厚，炎细胞浸润，肺泡腔内有少量红细胞漏出，偶见透明膜，IQA明显下降(P0.01)；S+PPG组肺损伤明显加重，肺泡塌陷及肺不张明显，多量肺泡见广泛透明膜形成，IQA升高(P0.05)。 3.血浆H_2S含量及肺组织CSE活性变化：与N组比较，S组血浆H_2S含量和肺组织CSE活性明显降低(P＜0.01)。与S组比较，S+L组血浆H_2S含量和肺组织CSE活性未见明显变化(P＞0.05)，S+M, S+H组血浆H_2S含量和肺组织CSE活性均显著升高(P＜0.01)；S+PPG组血浆H_2S含量及肺组织CSE活性降低(P＜0.01或P＜0.05)。 4.肺组织匀浆SOD及GSH-Px活力变化：与N组相比较，S组肺组织匀浆SOD及GSH-Px活力明显下降(P0.01)。与S组相比较，不认为S+L组肺组织匀浆SOD及GSH-Px活力升高存在差异(P0.05)，S+M, S+H组肺组织匀浆SOD及GSH-Px活力均有明显上升(P0.01)；S+PPG组肺组织匀浆SOD及GSH-Px活力减低(P0.05或P＜0.05)。 5.肺组织匀浆8-异构前列腺素(8-iso-PGF2α)含量的变化：与N组相比较，S组肺组织8-iso-PGF2α含量明显升高(P0.01)。与S组相比较，不认为S+L组肺组织8-iso-PGF2α含量下降存在差异(P0.05)，S+M, S+H组肺组织8-iso-PGF2α含量均有明显下降(P0.01)；S+PPG组肺组织8-iso-PGF2α含量升高(P0.01)。结论： 1.外源性补充H_2S，，蝮蛇毒染毒大鼠SOD与GSH-Px水平上升，8-isoPGF2a水平下降，肺损伤减轻；而抑制H_2S生成，蝮蛇毒染毒大鼠SOD与GSH-Px水平下降，8-isoPGF2a水平上升，肺损伤加重。 2. H_2S可能通过改善氧化/抗氧化系统之间的失衡状态，减轻蝮蛇毒染毒后大鼠急性肺损伤的程度。
[Abstract]:Purpose: The effect of H _ 2S/ CSE on oxidative stress in rats with acute lung injury was studied. For use. Methods:1. Group:36 male clean-grade SD rats were divided into 6 groups by a complete random design,6 rats in each group, respectively: the normal saline control group (N group), the control group (S group) and the low-dose NaHS group (group S). (S + L group); NaHS group (group S + M) in the high-dose NHS group (S + M group); the high-dose NHS group (S + H group); the pyotoxin + PPG group (S + PPG group).2. Method: The volume of physiological saline such as intraperitoneal injection of N groups and the rest of the group were divided into two groups. After 3 h, the volume of NS + L, S + M, S + H group and S + PPG group were injected intraperitoneally and the doses of 0.78 mg/ kg, 1.56 mg/ kg and 3.12 mg/ kg of NaHS and 30 mg/ k were respectively injected into the abdominal cavity. G-dose of PPG. After 6 h, each group of experimental animals was sacrificed, cardiac blood was collected and the serum was separated and the corresponding samples were taken. The lung tissue was retained as a specimen. The lung tissue HE staining was used to observe the pathological changes in parallel with the pathological changes, the wet/ dry weight ratio of the lung was measured, the plasma H _ 2S content was detected by the deproteinization method, the CSE activity of the lung tissue was determined by the methylene blue method, and the lung tissue homogenate superoxide dismutase was determined by the WST-1 method. The activity of glutathione peroxidase (GSH-Px) and the enzyme-linked immunosorbent assay (ELISA) were used to determine the activity of glutathione peroxidase (GSH-Px) in the lung tissue and the enzyme-linked immunosorbent assay (ELISA) for the determination of 8-isoprostane (8-iso-P) in the lung. GF2 1. Results:1. The change of the wet/ dry weight ratio (W/ D) of the lung tissue: compared with the N group, the S group W/ In comparison with S group, there was no difference in W/ D value of S + L group (P0.05), and W/ D of S + M and S + H group decreased significantly (P0.01); and S + PPG group W/ D. The pathological changes of the pulmonary tissue and the pathological score of the lung injury (IQA) under the light microscope: compared with the normal lung tissue of the N group, the pulmonary alveolar structure of the S group was obviously disturbed, destroyed, the alveolar structure was incomplete, the alveolar edema was obvious, and a large amount of the lung The formation of hyaline membrane, the marked thickening of the interstitial lung, the large amount of red blood cells and the infiltration of the pulmonary alveoli and the alveoli in the lung can be seen in the lung, and I can see that the In comparison with S group, there was no significant improvement in lung injury in S + L group (P0.05), and the lung injury in S + M and S + H group was relieved. There was a small amount of red blood cell leakage, and there was a marked decrease in IQA (P0.01); the lung injury in the S + PPG group was significantly increased, the alveolar collapse and the lung were not obvious, and the multi-dose alveoli were formed in the extensive hyaline membrane formation. IQA increased (P0.05).3. The plasma H _ 2S content and the changes of the activity of CSE in the lung tissue: compared with the N group, the plasma H _ 2S content in the S group and the lung tissue CS The activity of plasma H _ 2S and CSE in the S + L group was significantly higher than that of the S group (P <0.01). The plasma H _ 2S content in the S + group and the activity of CSE in the lung tissue were significantly increased (P <0.01), and the plasma H _ 2S content in the S + PPG group and the activity of the CSE in the lung tissue were decreased. (P <0.01 or P <0.05).4. SOD and GSH-Px activity in the lung tissue homogenate: compared with the N group, the SOD and GS of the lung tissue of the S group were homogenate. The activity of SOD and GSH-Px in the lung tissue of S + L group was significantly higher than that in S group (P0.05). The changes of the content of 8-iso-prostaglandin (8-iso-PGF2) in the lung tissue homogenate: compared with the N group, the S-group lung tissue 8-iso Compared with S group, there was no difference in the content of 8-iso-PGF2 in the lung of S + L group (P0.05), and the content of the 8-iso-PGF2 in the S + M, S + H group was significantly decreased (P0.01); and the lung tissue of the S + PPG group was 8-i. so- Conclusion:1. The level of SOD and GSH-Px in rats exposed to H _ 2S and the level of GSH-Px increased, and the level of 8-isoPGF2a decreased and the lung injury was reduced. A decrease in the level of 8-isoPGF2a and an increase in lung injury.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R563

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