基于抑制巨噬细胞氧化亢进，益气活血方防治门脉高压症的药理研究

Portal hypertension is a complication with an elevated portal pressure in digestive disorders. Yiqi-huoxue prescription is effective for the portal hypertension in clinic. Recently, its pathogenesis was related to the balance between NADPH oxidase 2 (NOX2) and superoxide dismutase 3 (SOD3) from the macrophages infiltrated in portal triads. We hypothesised that Yiqi-huoxue prescription effect on portal hypertension via inhibiting the oxidase hyperactivity of macrophages in portal triads. In this study, a pathogenic model of portal hypertension was established in rats with both CCl4-induced chronic hepatitis and macrophage depletion by liposomal clodronate. Then the rats with portal hypertension were treated with Yiqi-huoxue prescription. The special effect on the oxidase hyperactivity from macrophages in portal triads was confirmed with the potency of portal pressure in vivo and the biomarker of activated macrophages. The bioavailability of nitric oxide (NO) or prostacyclin I2 (PGI2) for dilating portal venules was evaluated in the isolated portal perfused rat liver with or without its special synthase inhibitor. The potency of NOX2/SOD3 activities in the macrophages next to terminal portal venules was exactly calculated with the cellular ratio in portal triads to that in total hepatic tissue, the genic dynamics of each enzyme was detected with its mRNA level by real time RT-PCR and its protein amount by western blot. The efficiency of Yiqi-huoxue prescription on portal hypertension in this pathogenic model was evaluated with the bioavailability of NO or PGI2, which be regulated by the potency of designated macrophages in portal triads, to exploit further portal hypertensive pathogenesis in basic and clinic. This work is going to exploit the special mechanisms of Yiqi-huoxue prescription, supporting the advantages of TCM in clinics.

门脉高压症是常见的难治疾病，病因多样，病机不清，益气活血方疗效满意。前期发现汇管区巨噬细胞降低门脉舒张物效应，课题组提出“抑制巨噬细胞氧化亢进是益气活血方防治门脉高压症”的药理机制。本课题对肝炎门脉高压症大鼠，氯膦酸脂质体清除巨噬细胞，形成模型配对大鼠，确认巨噬细胞激活致病。灌胃益气活血方防治“模型配对大鼠”，检测门脉压力与巨噬细胞激活标志，确认方剂疗效的病机特异性。在离体门脉灌流上，氨基胍或塞来昔布抑制合酶活性，观察门脉诱导性舒张收缩变化，分析一氧化氮或前列环素的利用度；以汇管区巨噬细胞“2型尼克酰胺腺嘌呤二核苷酸磷酸氧化酶与3型超氧化物歧化酶”基因表达、蛋白含量和酶促活性的效能比，确定抑制氧化亢进的平衡机制；递进分析血浆、肝脏与巨噬细胞的代谢物组差异，探索网络药效。本课题阐明了方剂防治门脉高压症的药理机制，有益于提高中医药优势病种诊治水平。

门脉高压症是常见的难治疾病，病机不清，益气活血方疗效满意。本课题对肝炎门脉高压症大鼠，氯膦酸脂质体清除巨噬细胞，形成模型配对大鼠，灌胃益气活血方治疗“模型配对大鼠”，检测门脉压力与巨噬细胞激活标志，确认方剂疗效。在离体门脉灌流上，分析一氧化氮或前列环素的利用度；以汇管区巨噬细胞“2型尼克酰胺腺嘌呤二核苷酸磷酸氧化酶(NOX2)与3型超氧化物歧化酶(SOD3)”基因表达、蛋白含量和酶促活性的效能比，确定抑制氧化亢进的平衡机制。结果发现汇管区巨噬细胞氧化亢进导致门脉压力升高；益气活血方有效单体甘草酸二铵、丹酚酸B可显著降低门脉压力，降低血清sCD163水平，减少巨噬细胞在汇管区的浸润，包括门脉舒张物一氧化氮和前列环素生物利用度；增加SOD3酶活性,降低NOX2和诱导型一氧化氮合酶表达，抑制巨噬细胞氧化亢进，治疗门脉高压症。为门脉高压症的临床防治提供了依据，并丰富了中医药治疗优势病种的内涵。