目的 通過構(gòu)建肝性腦病大鼠模型，探究安宮牛黃丸對肝性腦病大鼠腫瘤壞死因子α/核因子κB（TNF-α/NF-κB）信號通路及神經(jīng)功能的影響。方法 將60只SPF級SD大鼠分為對照組，模型組，乳果糖組（0.2 mL/200 g乳果糖）、安宮牛黃低劑量組（1.0 g/kg）、中劑量（2.0 g/kg）、高劑量（3.0 g/kg），每組10只。通過硫代乙酰胺（TAA）法構(gòu)建肝性腦病模型（連續(xù)3 d對模型大鼠注射300 mg/kg TAA溶液），造模前4 d開始各組每天連續(xù)ig相應(yīng)劑量乳果糖及安宮牛黃丸直至造模結(jié)束后2 d；對照組注射等量生理鹽水。水迷宮實驗檢測大鼠學(xué)習(xí)記憶能力；ELISA法檢測大鼠血清生化指標(biāo)及海馬組織乙酰膽堿（Ach）、谷氨酸（Glu）含量；蘇木精-伊紅染色（HE）觀察肝組織和腦組織病理學(xué)變化；實時熒光定量PCR檢測各組樣本中TNF-α、NF-κB mRNA的表達(dá)情況；Western blotting檢測通路相關(guān)蛋白表達(dá)情況。結(jié)果 與對照組相比，模型組尋臺潛伏期、血氨、ALT、AST水平、肝組織損傷、腦組織海馬CA1區(qū)損傷程度、Glu含量、TNF-α和NF-κB的mRNA及蛋白表達(dá)水平顯著增加，而穿臺次數(shù)、神經(jīng)學(xué)評分、Ach含量顯著降低（P<0.05）。與模型組相比，乳果糖組和安宮牛黃低、中、高劑量組大鼠尋臺潛伏期、血氨、ALT、AST水平、肝組織損傷、腦組織海馬CA1區(qū)損傷程度、Glu含量、TNF-α和NF-κB的mRNA及蛋白表達(dá)水平顯著降低，而穿臺次數(shù)、神經(jīng)學(xué)評分、Ach含量顯著增加（P<0.05），且安宮牛黃丸各組間存在一定劑量相關(guān)性。結(jié)論 安宮牛黃丸能抑制肝性腦病大鼠TNF-α/NF-κB信號通路，對大鼠神經(jīng)功能具有保護(hù)作用。

Objective To explore the effects of Angong Niuhuang Pill on tumor necrosis factor-α/nuclear factor-κB (TNF-α/NF-κB) signal pathway and neurological function in rats with hepatic encephalopathy by constructing a HE rat model. Methods Sixty SPF SD rats (10 rats in each group) were divided into control group, model group, lactulose group (0.2 mL/200 g lactulose), Angong Niuhuang low-dose group (1.0 g/kg), medium-dose group (2.0 g/kg), and high-dose group (3.0 g/kg). Hepatic encephalopathy model was established by thioacetamide (TAA) method (300 mg/kg TAA solution was injected into the model rats for 3 consecutive days). From 4 days before modeling, each treatment group was given a corresponding dose of lactulose and Angong Niuhuang Pill by gavage until 2 days after modeling; the control group was injected with the same amount of normal saline. Water maze test was used to test the ability of learning and memory, enzyme linked immunosorbent assay (ELISA) was used to detect the serum biochemical indexes and the contents of acetylcholine (ACh) and glutamic acid (Glu) in hippocampus, hematoxylin eosin staining (HE) was used to observe the pathological changes of liver and brain, real-time fluorescence quantitative PCR was used to detect the mRNA expressions of TNF-α and NF-κB, Western blot was used to detect the expression of pathway related proteins. Results Compared with the control group, the latency of platform searching, levels of blood ammonia, ALT, AST, the injury degree of liver tissue, injury degree of hippocampal CA1area in brain tissue, the content of Glu, the mRNA and protein expression levels of TNF-α and NF-κB in the model group were significantly increased, however, the times of crossing platform, neurological score and content of ACh were significantly decreased (P<0.05). Compared with those in model group, the latency of platform searching, the levels of blood ammonia, ALT, AST, the injury degree of liver tissue, injury degree of hippocampal CA1 area in brain tissue, the content of Glu, the mRNA and protein expression levels of TNF-α and NF-κB in lactulose group, low-dose group, medium-dose group and high-dose group were significantly decreased, however, the times of crossing platform, neurological score and content of ACh were significantly increased (P<0.05), there was a dose-dependent relationship between the Angong Niuhuang Pill groups. Conclusion Angong Niuhuang Pill can inhibit TNF-α/NF-κB signaling pathway in hepatic encephalopathy rats, and has protective effect on neurological function of rats.

R285.5

河南省中醫(yī)藥科學(xué)研究專項課題（2019YZ2052）