还原型谷胱甘肽对顺铂致大鼠急性肾损伤炎症反应的干预研究

发布时间：2018-05-12 03:23

本文选题：顺铂 + 急性肾损伤　；参考：《广西医科大学》2014年硕士论文

【摘要】：目的：通过腹腔注射顺铂建立大鼠急性肾损伤动物模型，检测血清肿瘤坏死因子-α（TNF-α）、C反应蛋白（CRP）、白细胞介素-6（IL-6）在顺铂致大鼠急性肾损伤中多个时间点的变化了解其炎症反应情况，同时探讨还原型谷胱甘肽对顺铂致大鼠急性肾损伤中上述炎症因子水平的影响以了解有否肾保护作用。方法：54只健康雄性SD大鼠为研究对象，根据随机数字法分为生理盐水对照组(CN组，n＝6)，顺铂模型组(CP组，n＝24)及还原型谷胱甘肽干预组(GI组，n＝24)，CP组为腹腔单次注射顺铂10mg/kg，根据给药后各时间点的不同分为CP24h组、CP48h组、CP72h组、CP96h组；GI组为应用顺铂前单次腹腔注射还原型谷胱甘肽800mg/kg，然后与CP组一样使用顺铂，根据顺铂给药时间不同分为GI24h组、GI48h组、GI72h组、GI96h组；每个时间点均为6只大鼠。CN组为腹腔注射与CP组等容量的生理盐水。取各时间点大鼠乙醚吸入麻醉，打开腹腔，门静脉取血检测血清肌酐(Scr)、TNF-α、CRP、IL-6，立即取出大鼠双肾，去除被膜，生理盐水冲洗后，沿冠状面分为两部分，用4%多聚甲醛固定过夜，拟行肾病理切片。结果：1.肾功能变化CP组在注射顺铂后24h血Scr较CN组升高不明显，差别无统计学意义（P＞0.05），48h Scr较CN组明显升高（P＜0.05），72h Scr较基线升高50%，提示顺铂致大鼠急性肾损伤建模成功；GI组上述时间点的Scr水平明显低于CP组（P＜0.05），略高于CN组无统计学差异（P＞0.05）； 2．炎症因子水平与CN组比较，CP组应用顺铂后24h TNF-α即有明显升高（P＜0.05），随之呈持续升高状态，至48h达高峰，72h、96h仍有较高水平；CP组应用顺铂后24h血CRP升高明显（P＜0.05），随后持续升高至72h达高峰，之后下降；CP组应用顺铂后72h血IL-6升高明显（P＜0.05），至96h仍有较高水平；GI组血TNF-α、CRP、IL-6有相似结果，但与CP组比较水平明显下降（P＜0.05），与CN组比较上述炎症因子水平仍有明显升高。 3．肾脏病理改变NS组大鼠肾组织HE染色可见：肾组织结构清晰未见明显异常，肾小球、肾小管结构正常，间质较少；CP组大鼠肾组织HE染色可见：除有淤积红细胞外肾小球未见明显异常，但肾小管明显变性，可见肾小管上皮细胞刷状缘脱落、胞质空泡变性、管腔扩张、肾小管堵塞、间质炎症细胞浸润，，甚至出现聚集凋亡细胞；且肾小管病变随时间逐渐加重。GI组大鼠肾小囊腔隙变窄，肾小管结构完整，细胞轻度变性、肿胀，少量管腔扩张，腔内偶见渗出物。结论：1.在顺铂致大鼠急性肾损伤中，血清炎症因子TNF-α、CRP、IL-6升高明显。 2.还原型谷胱甘肽对顺铂致大鼠急性肾损伤炎症反应有一定的保护作用。
[Abstract]:Objective: to establish a rat model of acute renal injury by intraperitoneal injection of cisplatin, and to detect the changes of serum tumor necrosis factor- 伪 (TNF- 伪) C-reactive protein (TNF- 伪) and interleukin-6 (IL-6) in acute renal injury induced by cisplatin in rats. At the same time, the effects of reduced glutathione on the level of inflammatory cytokines in acute renal injury induced by cisplatin in rats were investigated. Methods 54 healthy male Sprague-Dawley rats were studied. According to the random number method, the rats were divided into normal saline control group (CN group), cisplatin model group (CP group) and reduced glutathione intervention group (Glutathione intervention group). The control group was given a single intraperitoneal injection of 10 mg / kg cisplatin. According to the different time points after administration, they were divided into CP24h group (CP48h group) and reduced glutathione intervention group (Glutathione intervention group). Glutathione (800 mg / kg) was injected intraperitoneally before cisplatin was used in GI group, and then cisplatin was used as CP group. According to the time of cisplatin administration, the rats were divided into GI24h group (Gi 48 h) and Gi 72 h group (Gi 96 h group), 6 rats in each group were intraperitoneally injected with normal saline of the same volume as CP group. The rats were anesthetized by ether inhalation at different time points, the abdominal cavity was opened, and the blood samples from portal vein were taken out to detect the serum creatinine, TNF- 伪 and CRP IL-6. The bilateral kidneys of the rats were removed immediately, the membrane was removed. After rinsing with normal saline, the rats were divided into two parts along the coronal surface and fixed with 4% paraformaldehyde for the night. Pathological sections of the kidney were prepared. The result is 1: 1. The changes of renal function in CP group were not significantly higher than those in CN group at 24 h after injection of cisplatin. There was no significant difference in Scr between CN group and CP group (P > 0.05) at 48h. The level of Scr in GI group was significantly lower than that in CP group (P < 0.05), and there was no significant difference between CN group and CN group (P > 0.05). The results showed that the level of Scr in GI group was significantly lower than that in CP group (P < 0.05), and it was slightly higher than that in CN group (P > 0.05). 2. Compared with CN group, the level of inflammatory cytokines in CP group increased significantly 24 hours after cisplatin administration (P < 0.05), and continued to rise, and reached the peak at 48h to 72h / 96h. There was still a higher level of CRP in CP group 24h after cisplatin administration (P < 0.05), and then it continued to rise to the peak at 72 h (P < 0.05). The level of TNF- 伪 in CP group was significantly higher than that in CP group (P < 0.05). After that, the serum IL-6 increased significantly 72 hours after treatment with cisplatin in the CP group (P < 0.05), and there was a similar result at 96 h in the group with high level of TNF- 伪 CRPU IL-6, but the level was significantly lower than that in the CP group (P < 0.05), and the level of the above inflammatory cytokines was still significantly higher than that in the CN group. 3. The pathological changes of kidney in NS group showed that the clear structure of renal tissue was not abnormal, glomeruli and tubule structure were normal. The HE staining in renal tissue of rats with less interstitial protein showed that there was no obvious abnormality in glomeruli except for the accumulation of red blood cells, but the renal tubules were obviously denatured, the brush edge of renal tubule epithelial cells fell off, the cytosolic vacuoles degenerated, the lumen dilated, the renal tubules were blocked, and the renal tubules were blocked. The interstitial inflammatory cells infiltrated and even apoptotic cells appeared, and the renal tubule lesion gradually increased with the time. GI group, the renal tubule structure was intact, the cells were slightly denatured, swelling, a small amount of lumen dilatation. Exudates were occasionally seen in the cavity. Conclusion 1. In the acute renal injury induced by cisplatin, the serum inflammatory factor TNF- 伪 was significantly increased. 2. Reduced glutathione can protect acute renal injury induced by cisplatin in rats.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R730.53;R692.5

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