摘要
目的探究盐酸纳美芬对缺氧缺血性脑病(HIE)新生大鼠Ras相关C3肉毒素底物1(Rac1)/烟酰胺腺嘌呤二核苷酸磷酸氧化酶2(NOX2)通路及神经细胞凋亡的影响。方法构建HIE新生大鼠模型,使用随机数字表法分为模型组,盐酸纳美芬低、中、高剂量(10μg/kg、15μg/kg、20μg/kg,尾静脉注射)组,地塞米松组(阳性对照,6 mg/kg,腹腔注射),另取假手术组新生大鼠,每组20只,模型组和假手术组大鼠给予等剂量生理盐水,其余各组给予相对应剂量药物,每天1次,连续5 d。干湿比重法检测新生大鼠脑组织含水率;2,3,5-三苯基氯化四氮唑(TTC)染色法检测新生大鼠脑梗死面积;苏木精-伊红(H-E)染色法观察新生大鼠海马组织病理学变化;酶联免疫吸附法(ELISA)检测新生大鼠海马组织白细胞介素-1β(Interleukin-1β,IL-1β)、肿瘤坏死因子-α(Tumor necrosis Factor-α,TNF-α)和白细胞介素-6(Interleukin-6,IL-6)含量;TUNEL染色检测新生大鼠神经细胞凋亡指数;蛋白免疫印迹法(Western blot)法检测新生大鼠海马组织Rac1和NOX2蛋白表达水平。结果假手术组大鼠海马组织结构正常,未见明显病理损伤。与假手术组相比,模型组大鼠海马组织神经元体积增大,间质水肿,脑组织含水率、脑梗死面积、神经细胞凋亡指数、IL-1β含量、TNF-α含量、IL-6含量及Rac1和NOX2蛋白表达水平显著升高(P<0.05);与模型组相比,盐酸纳美芬低、中、高剂量组大鼠海马组织病理损伤依次缓解,脑组织含水率、脑梗死面积、神经细胞凋亡指数、IL-1β含量、TNF-α含量、IL-6含量及Rac1和NOX2蛋白表达水平依次降低(P <0.05);盐酸纳美芬高剂量组与地塞米松组新生大鼠各项指标差异无统计学意义(P>0.05)。结论盐酸纳美芬可能通过抑制Rac1/NOX2通路,减轻HIE新生大鼠海马组织炎症反应,减少神经细胞凋亡。
Objective To investigate the effects of nalmefene hydrochloride on Ras-related C3 botulinum toxin substrate 1(Rac1)/nicotinamide adenine dinucleotide phospho oxidase 2(NOX2) pathway and neuronal apoptosis in neonatal rats with hypoxic ischemic encephalopathy(HIE). Methods HIE neonatal rat models were established and randomly divided into model group, low, medium and high dose nalmefene hydrochloride groups(10 μg/kg, 15 μg/kg, 20 μg/kg, tail vein injection)and dexamethasone group(positive control, 6 mg/kg, intraperitoneal injection), another neonatal rats in sham operation group were selected, with 20 rats in each group. Rats in model group and sham operation group were given equal dose of normal saline, and the other groups were given the corresponding dose of drugs, once a day for 5 consecutive days. The water content of brain tissue, the infarct area and the pathological changes of hippocampus in neonatal rats was measured by dry-wet specific gravity method, 2,3,5-triphenyl tetrazolium chloride(TTC) staining and hematoxylin-eosin(H-E) staining respectively;the contents of interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) were detected by enzyme-linked immunosorbent assay(ELISA);TUNEL staining was used to detect neuronal apoptosis index;Western blot was used to detect the protein expression levels of Rac1 and NOX2 in hippocampus of neonatal rats. Results In the sham operation group, the hippocampal structure was normal without obvious pathological damage. Compared with those in the sham operation group,the volume of hippocampal neurons in the model group was higher and the interstitial edema was observed, brain water content, cerebral infarction area, neuronal apoptosis index, IL-1β, TNF-α, IL-6 contents and Rac1 and NOX2 protein expression levels were significantly higher(P<0.05);compared with those in the model group, the pathological damage of hippocampus in the low, medium and high dose nalmefene hydrochloride groups was alleviated in turn, brain water content, cerebral infarction area, neuronal apoptosis index, IL-1β, TNF-α, IL-6 contents and Rac1 and NOX2 protein expression levels decreased in turn(P<0.05);there was no significant difference between the high dose nalmefene hydrochloride group and dexamethasone group(P>0.05). Conclusion Nalmefene hydrochloride may reduce the inflammatory reaction and neuronal apoptosis in the hippocampus of neonatal rats with HIE by inhibiting Rac1/NOX2 pathway.
作者
白鹏祖
张利亮
张海盛
贾珍
BAI Pengzu;ZHANG Liliang;ZHANG Haisheng;JIA Zhen(Department of Anesthesiology,Qinghai Red Cross Hospital,Xining,Qinghai 810000,China;Department of Anesthesiology,Affiliated Hospital of Qinghai University,Xining,Qinghai 810000,China)
出处
《中国优生与遗传杂志》
2021年第7期955-960,共6页
Chinese Journal of Birth Health & Heredity
