血必净注射液对大鼠下肢缺血再灌注损伤的影响

发布时间：2018-01-22 08:36

本文关键词： 血必净注射液缺血-再灌注损伤下肢骨骼肌　出处：《山西医科大学》2012年硕士论文　论文类型：学位论文

【摘要】：目的：建立大鼠下肢缺血-再灌注损伤（ischemia-reperfusion injure, IRI）动物模型，再灌注即刻给予血必净注射液，初步探讨血必净对大鼠下肢缺血-再灌注损伤中的影响，为进一步开展临床上应用血必净防治肢体缺血-再灌注损伤奠定基础。方法：健康Wistar大鼠36只，随机分为三组，每组12只。（1）假手术组（sham operationgroup，SOG组），开腹只分离腹主动脉，不夹闭腹主动脉。（2）缺血再灌注组（I/R组），无创动脉夹夹闭腹主动脉2h后，再灌注即刻自尾静脉注射生理盐水（8ml/kg），松开血管夹恢复血流灌注4h。（3）血必净组（XBJ组），夹闭腹主动脉2h，再灌注即刻自尾静脉注射血必净注射液（8ml/kg），松开血管夹恢复血流灌注4h。每组动物分别在再灌注4h后抽取血液标本，分离血清，测定血清中MPO、LDH、TNOS、iNOS和cNOS的活性以及HE染色后光镜下观察骨骼肌、血管壁结构的变化。结果：于SOG组比较，I/R组XBJ组血清中MPO、LDH、TNOS、iNOS的活性均增高（P0.05），I/R中血清中cNOS活性降低（P0.05），XBJ组血清中cNOS活性增高（P0.05）。于I/R组比较，XBJ组血清中MPO、LHD、iNOS活性均降低（P0.05），XBJ组血清中TNOS、cNOS活性增高（P0.05）。I/R组再灌注4小时后骨骼肌细胞肿胀；细胞间隙变窄，大量肌细胞断裂，细胞间隙中有大量炎性细胞浸润。肌纤维水肿，排列不整齐，部分肌纤维有断裂；血管壁各层排列疏松，细胞水肿，VECs胞核向管腔突出明显，VECs与内皮下弹性膜结合松散，部分细胞脱落至管腔内，部分血管内皮细胞缺失，可见炎性细胞黏附。XBJ组再灌注4小时骨骼肌细胞肿胀减轻，细胞间隙趋于正常，肌细胞断裂较少，细胞间隙中炎性细胞浸润减少，细胞核肿胀减轻；肌纤维排列较整齐，肌纤维断裂较少；VECs排列尚规则，部分细胞水肿，与内皮下弹性膜结合较紧密，内皮细胞脱落及炎性细胞粘附较少结论：实验表明血必净注射液对缺血再灌注损伤有一定的保护作用。其保护作用主要表现在稳定中性粒细胞，降低iNOS活性，减少iNOS源性NO；增高cNOS活性，使eNOS源性NO增加，，减少对细胞的毒性作用，减轻细胞水肿，降低细胞膜通透性，恢复细胞的正常功能。
[Abstract]:Objective: to establish an animal model of ischemia-reperfusion injury (IRI) in rats and to give Xuebijing injection immediately after reperfusion. To explore the effect of Xuebijing on lower limb ischemia-reperfusion injury in rats, and to lay a foundation for further clinical application of Xuebijing in prevention and treatment of limb ischemia-reperfusion injury. Methods: Thirty-six healthy Wistar rats were randomly divided into three groups (12 rats in each group) sham operationg roupSOG group (sham operation group). The abdominal aorta was isolated from the abdominal aorta without clamping the abdominal aorta in the ischemia reperfusion group. The abdominal aorta was clamped by non-invasive artery for 2 hours. Immediately after reperfusion, 8 ml / kg of normal saline was injected into the caudal vein, and the blood flow perfusion was restored 4 h. 3) in XBJ group, the abdominal aorta was clipped for 2 hours. Immediately after reperfusion, Xuebijing injection was injected into caudal vein at 8 ml / kg 路kg ~ (-1), and blood perfusion was recovered for 4 h by loosening the clip. Blood samples were taken from each group for 4 hours after reperfusion and serum was isolated. The activities of iNOS and cNOS in serum were measured and the changes of skeletal muscle and vascular wall structure were observed under light microscope after HE staining. Results: compared with SOG group, the activity of XBJ in XBJ group was significantly higher than that in XBJ group (P 0.05). The activity of cNOS in the serum of I / R group was lower than that in the group of P0.05 and XBJ, and the activity of cNOS in the serum of the group of P0.05 was significantly higher than that in the group of I / R, compared with that of the group of XBJ in the I / R group. The activity of iNOS decreased in both groups. The activity of TNOS-CNOS increased in the serum of P0.05 + XBJ group and the swelling of skeletal muscle cells was observed in P0.05 + I / R group after 4 hours of reperfusion. The intercellular space became narrow, a large number of muscle cells broke, a large number of inflammatory cells infiltrated in the intercellular space, the muscle fibers were edema, disordered arrangement, and some of the muscle fibers were broken. The layers of vascular wall were loosely arranged, and the VECs nucleus protruded into the lumen. The VECs were loosely bound to the endodermic elastic membrane, some of the cells fell off into the lumen and some of the vascular endothelial cells were missing. It can be seen that the inflammatory cell adhesion. XBJ group reduced the swelling of skeletal muscle cells at 4 hours after reperfusion, the gap tended to be normal, the break of muscle cells was less, the infiltration of inflammatory cells in the intercellular space was decreased, and the swelling of nucleus was alleviated. The arrangement of muscle fibers was neat, and the breakage of muscle fibers was less. The VECs arrangement is regular, some cells are edema, binding to the endodermic elastic membrane is close, endothelial cells fall off and inflammatory cell adhesion is less. Conclusion: Xuebijing injection has a certain protective effect on ischemia-reperfusion injury. The protective effect of Xuebijing injection is mainly in stabilizing neutrophil, decreasing iNOS activity and decreasing iNOS derived no. By increasing the activity of cNOS, the activity of eNOS derived no was increased, the toxic effect of eNOS on cells was reduced, the edema of cells was alleviated, the permeability of cell membrane was decreased, and the normal function of cells was restored.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R-332

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