黄芩苷对脓毒症小鼠急性肾损伤的保护作用及机制研究  

Protective effect and mechanism of Baicalin on acute kidney injury in septic mice

作　　者：乐健伟[1] 樊恒[1] 孙敏[1] 朱建华[1] Le Jianwei;Fan Heng;Sun Min;Zhu Jianhua(Department of Intensive Care Unit,Ningbo First Hospital,Ningbo 315010,Zhejiang,China)

机构地区：[1]宁波市第一医院重症医学科,浙江宁波315010

出　　处：《中华危重病急救医学》2021年第7期866-870,共5页Chinese Critical Care Medicine

基　　金：浙江省医药卫生科技计划项目(2020KY815,2020KY828,2019ZA113);浙江省宁波市科技计划项目(2018A610296)。

摘　　要：目的探讨不同剂量黄芩苷(BAI)对脓毒症小鼠急性肾损伤(AKI)的保护作用及机制。方法按随机数字表法将100只实验小鼠分为假手术组(Sham组)、盲肠结扎穿孔术(CLP)致脓毒症模型组(CLP组)和BAI预处理组,其中BAI预处理组根据BAI用量分为低、中、高剂量组(BAI-L+CLP、BAI-M+CLP、BAI-H+CLP组),每组20只。通过CLP法制备小鼠脓毒症相关性急性肾损伤(SA-AKI)模型;Sham组仅开腹、关腹,不予盲肠结扎、穿孔。BAI预处理组小鼠每日分别灌胃BAI 25、50、100 mg/kg,持续3 d,第3天灌胃结束后6 h行CLP制备脓毒症模型;Sham组及CLP组灌胃等量蒸馏水。各组分别于术后24 h随机处死10只小鼠取眼眶血,采用酶联免疫吸附试验(ELISA)测定肾功能〔包括血肌酐(SCr)、血尿素氮(BUN)、血中性粒细胞明胶酶相关载脂蛋白(pNGAL)及血肾损伤分子-1(pKIM-1)〕;取肾组织,苏木素-伊红(HE)染色后,光镜下观察肾组织损伤情况,同时采用原位末端缺刻标记法(TUNEL)观察肾小管上皮细胞凋亡情况;采用蛋白质免疫印迹试验(Western blotting)测定肾组织细胞FLICE样抑制蛋白(c-FLIP)表达水平。各组其余10只小鼠用于统计术后7 d生存情况。结果CLP组小鼠肾小管上皮细胞大量变性、坏死,肾小管管腔明显扩张,间质内可见炎性细胞广泛浸润,肾功能急剧恶化。与CLP组比较,给予低剂量BAI预处理后,小鼠肾功能有所改善,但各指标差异无统计学意义;给予中、高剂量BAI预处理后,小鼠肾功能明显改善,SCr、BUN、pNGAL和pKIM-1均显著降低〔SCr(μmol/L):135.16±5.18、125.70±5.26比170.42±5.42,BUN(mmol/L):33.59±1.77、27.29±1.61比45.68±1.39,pNGAL(μg/L):91.29±4.68、73.40±3.77比131.50±6.55,pKIM-1(μg/L):6.34±0.30、5.51±0.35比8.03±0.29,均P<0.01〕,肾组织病理损伤显著减轻,肾小管上皮细胞凋亡数量明显减少(个/HP:16.20±0.49、13.10±0.66比29.60±0.49,均P<0.01),肾组织c-FLIP蛋白表达水平显著升高〔c-FLIP�Objective To observe the protective effect and mechanism of different doses of Baicalin(BAI)on acute kidney injury(AKI)in septic mice.Methods According to the random number table,100 mice were divided into sham operation group(Sham group),cecal ligation and perforation(CLP)induced sepsis model group(CLP group)and BAI pretreatment groups.The mice in BAI pretreatment groups were divided into low-,medium-and high-dose groups(BAI-L+CLP,BAI-M+CLP,BAI-H+CLP groups),with 20 mice in each group.A murine sepsis associated-acute kidney injury(SA-AKI)model was reproduced using CLP.The mice in the Sham group were only opened and closed the abdomen,without ligating or perforating the cecum.The mice in the BAI pretreatment groups were given BAI 25,50 and 100 mg/kg daily for 3 days,and CLP was performed at 6 hours after administration of BAI at the 3rd day to reproduce sepsis model.The mice in the Sham group and CLP group were given the same amount of distilled water as control.Ten mice were sacrificed at 24 hours after operation to collect orbital blood for renal function determination[serum creatinine(SCr),blood urea nitrogen(BUN),plasma neutrophil gelatinase-associated lipocalin(pNGAL)and plasma kidney injury molecule-1(pKIM-1)]by enzyme linked immunosorbent assay(ELISA).The kidney tissue was collected to observe the kidney tissue injury under light microscope after hematoxylin-eosin(HE)staining.The TdT-mediated dUTP nick-end labeling(TUNEL)was used to detect the apoptosis of renal tubular epithelial cells.Western blotting was used to detect the expression of cell FLICE like inhibitory protein(c-FLIP)in renal tissue.The remaining 10 mice in each group were used to calculate the survival rate of 7 days after operation.Results The renal tubular epithelial cells in the CLP group were massively degenerated with necrosis,the renal tubular lumen was significantly expanded,and inflammatory cells were widely infiltrated in the renal interstitium.Furthermore,the renal function deteriorated rapidly.Compared with the CLP group,the renal

关 键 词：黄芩苷 脓毒症 急性肾损伤 小鼠 

分 类 号：R285.5[医药卫生—中药学]