摘要
研究蝉花虫草提取物N^6-(2-羟乙基)腺苷[N^6-(2-hydroxyethyl)-adenosine,HEA]对小鼠肾脏缺血再灌注损伤(ischemia reperfusion,IR)的影响。选择20–25g雄性C57BL/6小鼠,随机分成5组。假手术组小鼠仅接受腹中线开腹、游离双侧肾蒂及缝合腹部操作;IR组小鼠制成肾脏IR模型,不给药;HEA低剂量组、HEA中剂量组、HEA高剂量组分别在建立肾脏缺血再灌注模型前15min腹腔注射HEA(2.5mg/kg、5mg/kg和7.5mg/kg)。再灌注24h后检测各组肾功能指标血清肌酐(serum creatinine,Scr)和尿素氮(blood urea nitrogen,BUN)水平。苏木精-伊红染色后用光学显微镜观察肾脏组织形态学改变。电镜和TUNEL分析法观察肾小管上皮细胞凋亡状况。实时荧光定量RT-PCR检测细胞间粘附分子-1(intercellular adhesion molecule-1,ICAM-1)、白细胞介素(interleukin-1β,IL-1β)和肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)等基因的m RNA的表达情况。结果显示,与假手术组比较,IR组BUN和Scr水平明显升高(P<0.01,P<0.05);肾脏病理表现为上皮细胞碎片和管型,且最为严重;与IR组比较,HEA 7.5mg/kg组血清BUN和Scr水平显著降低(P<0.05);HEA 5mg/kg组和HEA 7.5mg/kg组肾损伤情况明显改善(P<0.05,P<0.01);电镜观察可知HEA 7.5mg/kg组明显改善肾小管上皮细胞的损伤程度,并且保护线粒体和细胞核膜的完整性。原位末端标记法[terminal dexynucleotidyl transferase(Td T)-mediated d UTP nick end labeling,TUNEL]显示HEA治疗组凋亡细胞数量明显低于IR组(P<0.01)。HEA治疗组ICAM-1、IL-1β和TNF-αm RNA的水平均低于IR组(P<0.01)。所以预处理HEA 7.5mg/kg对肾脏缺血再灌注有保护作用。
The protective effects of N6-（2-hydroxyethyl）-adenosine （HEA） extracted from Ophiocordyceps sobolifere on rena schemia reperfusion injury （IRI） in mice were investigated. C57BL/6 mice were randomly divided into 5 groups, IR （ischemia reperfusion） group, sham-operated （SO） group, low-dose, middle-dose and high-dose HEA groups. The mice were injected with 2.5mg/kg, 5mg/kg and 7.5mg/kg HEA just as 25 minutes before ischemia reperfusion. In 24h after reperfusion, the serum and renal samples were collected. The serum creatinine （Scr） and blood urea nitrogen （BUN） levels in serum were determined for nvestigating the index of renal function. Renal patho-histological changes were also examined by HE staining. The condition of the apoptosis of renal tubular epithelial cells was observed by electron microscopy and TUNEL. Expressions of intercellular adhesion molecule-2 （ICAM-1）, interleukin-213 （IL-1β） and tumor necrosis factor-a （TNF-a） were assessed by using quantitative fluorescence RT-PCR. The result showed that as compared with the sham operation group, the BUN and Scr levels of IR group were significantly e Compared with the evated （P〈0.01, P〈0.05）. Renal pathology showed epithelial cell debris and tube types were the most severe R group, HEA 7.5mg/kg group exhibited lower levels of Scr and BUN （P〈0.05）. HEA 5mg/kg group and HEA 7.5mg/kg group significantly improved renal injury （P〈0.05, P〈0.02）. Electron microscopic observation indicated that the HEA 7.5mg/kg group significantly reduced the degree of damage of renal tubular in ischemia reperfusion injury, especially on mitochondrial and cell membrane. TUNEL （terminal dexynucleotidyl transferase（TdT）-mediated dUTP nick end labeling） staining analysis indicated that the number of apoptotic cells in HEA pretreatment group was much smaller than that in IR group （P〈0.01） The level of ICAM-2, IL-1β and TNF-a mRNA of HEA treatment group was lower than that of IR group （P〈0.02）. Therefore, pretreatment with HEA （7.5mg/kg） had protective effects on renal ischemia reperfusion injury in mice
出处
《菌物学报》
CAS
CSCD
北大核心
2015年第2期311-320,共10页
Mycosystema
基金
浙江省重大优先项目(2008C12044-3)
浙江省自然科学基金(Y3100625)
