茵陈水提物对新生豚鼠高胆红素血症耳蜗核抗炎保护作用

张奕　吴小海　林成光　艾力　杨应浩　冯桂明　黄旭龙　陈开贤　钟小苑

【摘要】 目的：研究茵陈水提物对新生豚鼠高胆红素血症的耳蜗核凋亡保护作用。方法：将50只3日龄新生豚鼠随机分成5组，每组10只，除了正常组，其余各组腹腔胆红素溶液100 μg/g，给药前和给药后1、4 h分别进行ABR动态监测。模型组给予2 mL等体积的生理盐水，其余分别设茵陈水提物高中低[8、4、2 g/（kg·d）]灌胃，共给药14 d，分别检查ABR值，试剂盒检测血清胆红素含量，荧光定量PCR检测TNF-α、IL-1β mRNA，ELSIA检测血清炎症因子TNF-α、IL-1β表达。结果：高胆红素改变ABR值主要表现在Ⅲ波、Ⅴ波、Ⅰ～Ⅲ波间延长。在模型组，炎症因子TNF-α和IL-1β基因水平和蛋白水平与正常组比较均明显升高（F=1 021.2，P=0.021）。结论：高胆红素改变ABR值的敏感变化，茵陈提取物有效预防胆红素对新生豚鼠的炎症因子升高，预防高胆红素对听觉神经的损伤。

【关键词】 茵陈水提物; 胆红素; 新生豚鼠; ABR; TNF-α; IL-1β

【Abstract】 Objective：To study the protective effect of Yinchen water extract on cochlear nuclear apoptosis in neonatal guinea pigs with hyperbilirubinemia.Method：A total of 50 three-day-old newborn guinea pigs were randomly divided into 5 groups，10 in each group.Except the normal group，100 μg/g peritoneal bilirubin solution and ABR were detected before administration and 1 h and 4 h after administration.The model group was given 2 mL equal volume of normal saline，and the rest were respectively treated with Yinchen water extract[8，4，and 2 g/（kg·d）]，and they were administered for 12 days.The serum bilirubin level was measured by the kit.The expression of serum inflammatory factors（TNF-α and IL-1β）were detected by Real-time PCR.ELSIA was used to detect TNF-α and IL-1β mRNA.Result：The ABR value of hyper bilirubin was mainly expressed in the Ⅲ wave，Ⅴ wave and Ⅰ-Ⅲ wave.In the model group.The inflammatory factors TNF-α and IL-1β gene levels and protein levels were significantly higher than those in the normal group（F=1 021.2，P<0.05）.Conclusion：Hyper bilirubin changes the sensitive change of ABR value.Yinchen water extract can effectively prevent the increase of inflammatory factors of bilirubin in newborn guinea pigs.

【Key words】 Yinchen water extract; Bilirubin; Newborn guinea pig; ABR; TNF-α; IL-1β

First-authors address：The Second Affiliated Hospital，Guangdong Pharmaceutical University，Guangzhou 510300，China

doi：10.3969/j.issn.1674-4985.2019.09.003

新生兒血清总胆红素水平大于221 μmol/L被定义为新生儿高胆红素血症[1]，其发病率占住院新生儿的30%～50%[2]，经常并发胆红素脑病，导致脑中枢神经系统损伤，引起运动功能不全、视觉听觉受损及智力低下等多种神经系统后遗症，甚至死亡，造成家庭社会的沉重负担[3-5]。研究表明听力损失主要发生在血清总胆红素水平中度增高时（TSB>1 768 μmol/L）[6]。认识胆红素脑病发病机制，寻找利于早期识别和干预新生儿高胆红素血症，重点预防胆红素对听力系统的破坏势在必行。本研究通过建立高胆红素血症模型，研究茵陈水提物干预后低位中枢核团细胞炎症因子的表达关系，探讨茵陈水提物对新生豚鼠高胆红素耳蜗核保护作用机制，为临床疾病防治提供理论依据。现报道如下。

1 材料与方法

1.1 动物 选用出生3 d的健康豚鼠50只，由南方医科大学实验动物中心提供，体重60～80 g，雌雄各半，耳廓反射灵敏，无强噪声暴露及耳毒性药物使用史，ABR测试正常。动物实验通过广东药科大学动物伦理委员会审查。

1.2 仪器与试剂配制 ABR仪器：美国Intelligent hearing systems，分析天平：上海越平仪器，减压真空泵：郑州科学仪器，离心机：长沙湘仪，酶标仪：瑞士迪肯，荧光定量PCR仪器，ABI7500。胆红素溶液配制：避光称取晶体胆红素50 mg，溶于1 mL 0.5 mol/L NaOH中，加入双蒸水9 mL，用0.5 mol/L HCl调节pH值至8.5。0.22 μm滤膜过滤，4 ℃避光保存。茵陈蒿汤：茵陈200 g，3 L水浸泡2 h，先文火后武火，煎煮2次，每次30 min，布氏漏斗真空过滤，合并2次所得滤液，真空减压浓缩至100 mL，浓度为含原药材量2 g/mL，4 ℃保存备用。