茵陳水提物對新生豚鼠高膽紅素血癥耳蝸核抗炎保護作用

張奕　吳小?！×殖晒狻“Α顟啤●T桂明　黃旭龍　陳開賢　鐘小苑

【摘要】 目的：研究茵陳水提物對新生豚鼠高膽紅素血癥的耳蝸核凋亡保護作用。方法：將50只3日齡新生豚鼠隨機分成5組，每組10只，除了正常組，其余各組腹腔膽紅素溶液100 μg/g，給藥前和給藥后1、4 h分別進行ABR動態(tài)監(jiān)測。模型組給予2 mL等體積的生理鹽水，其余分別設茵陳水提物高中低[8、4、2 g/（kg·d）]灌胃，共給藥14 d，分別檢查ABR值，試劑盒檢測血清膽紅素含量，熒光定量PCR檢測TNF-α、IL-1β mRNA，ELSIA檢測血清炎癥因子TNF-α、IL-1β表達。結(jié)果：高膽紅素改變ABR值主要表現(xiàn)在Ⅲ波、Ⅴ波、Ⅰ～Ⅲ波間延長。在模型組，炎癥因子TNF-α和IL-1β基因水平和蛋白水平與正常組比較均明顯升高（F=1 021.2，P=0.021）。結(jié)論：高膽紅素改變ABR值的敏感變化，茵陳提取物有效預防膽紅素對新生豚鼠的炎癥因子升高，預防高膽紅素對聽覺神經(jīng)的損傷。

【關鍵詞】 茵陳水提物; 膽紅素; 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

新生兒血清總膽紅素水平大于221 μmol/L被定義為新生兒高膽紅素血癥[1]，其發(fā)病率占住院新生兒的30%～50%[2]，經(jīng)常并發(fā)膽紅素腦病，導致腦中樞神經(jīng)系統(tǒng)損傷，引起運動功能不全、視覺聽覺受損及智力低下等多種神經(jīng)系統(tǒng)后遺癥，甚至死亡，造成家庭社會的沉重負擔[3-5]。研究表明聽力損失主要發(fā)生在血清總膽紅素水平中度增高時（TSB>1 768 μmol/L）[6]。認識膽紅素腦病發(fā)病機制，尋找利于早期識別和干預新生兒高膽紅素血癥，重點預防膽紅素對聽力系統(tǒng)的破壞勢在必行。本研究通過建立高膽紅素血癥模型，研究茵陳水提物干預后低位中樞核團細胞炎癥因子的表達關系，探討茵陳水提物對新生豚鼠高膽紅素耳蝸核保護作用機制，為臨床疾病防治提供理論依據(jù)?，F(xiàn)報道如下。

1 材料與方法

1.1 動物 選用出生3 d的健康豚鼠50只，由南方醫(yī)科大學實驗動物中心提供，體重60～80 g，雌雄各半，耳廓反射靈敏，無強噪聲暴露及耳毒性藥物使用史，ABR測試正常。動物實驗通過廣東藥科大學動物倫理委員會審查。

1.2 儀器與試劑配制 ABR儀器：美國Intelligent hearing systems，分析天平：上海越平儀器，減壓真空泵：鄭州科學儀器，離心機：長沙湘儀，酶標儀：瑞士迪肯，熒光定量PCR儀器，ABI7500。膽紅素溶液配制：避光稱取晶體膽紅素50 mg，溶于1 mL 0.5 mol/L NaOH中，加入雙蒸水9 mL，用0.5 mol/L?HCl調(diào)節(jié)pH值至8.5。0.22 μm濾膜過濾，4 ℃避光保存。茵陳蒿湯：茵陳200 g，3 L水浸泡2 h，先文火后武火，煎煮2次，每次30 min，布氏漏斗真空過濾，合并2次所得濾液，真空減壓濃縮至100 mL，濃度為含原藥材量2 g/mL，4 ℃保存?zhèn)溆谩?/p>