三甲益肝沖劑對實(shí)驗(yàn)性肝纖維化大鼠脂質(zhì)過氧化的影響

【摘要】  　　目的 觀察三甲益肝沖劑對大鼠肝纖維化的作用并探討與抗脂質(zhì)過氧化有關(guān)的作用機(jī)制。方法 雄性SD大鼠40只，隨機(jī)分為模型組、復(fù)方丹參組、三甲益肝沖劑組和正常組。以二甲基亞硝胺(DMN)腹腔注射誘導(dǎo)大鼠肝纖維化模型，復(fù)方丹參組造模同時給予復(fù)方丹參灌胃，三甲益肝沖劑組造模同時給予三甲益肝沖劑灌胃，共4周。4周后處死大鼠取肝組織標(biāo)本，光鏡觀察肝組織的病理變化，放射免疫法測定血清丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性。結(jié)果 與正常組比較，模型組肝組織Masson染色膠原纖維面積和血清MDA含量顯著增加(P<0.05)，而血清SOD活性明顯下降(P<0.05)；與模型組比較，三甲益肝沖劑組Masson染色膠原纖維面積和血清MDA含量顯著下降(P<0.05)，而SOD活性明顯升高(P<0.05)。Masson 染色膠原纖維面積和血清MDA含量表達(dá)呈正相關(guān)關(guān)系(r=0.435, P<0.05)，與SOD活性呈負(fù)相關(guān)關(guān)系(r=-0.358, P<0.05)。結(jié)論 三甲益肝沖劑具有良好的抗實(shí)驗(yàn)性大鼠肝纖維化作用，其主要作用機(jī)制與抗肝臟脂質(zhì)過氧化損傷有關(guān)。

【關(guān)鍵詞】  肝纖維化；三甲益肝沖劑；肝星狀細(xì)胞；脂質(zhì)過氧化通信論文發(fā)表
　　
　　ABSTRACT: Objective  To observe the effects of Sanjia Yigan Granule (S�睯G) on hepatic fibrosis and explore its mechanism related to anti�瞝ipid peroxidation in rats. Methods  Forty male SD rats were randomly divided into four groups: normal group, model group, and compound Salviae Miltiorrhizae (Sm) group and S�睯G group. Dimethylnitrosamine was injected intraperitoneally for 4 weeks to induce hepatic fobrosis. At the same time of modeling, Sm and S�睯G were given in the corresponding groups. The rats were killed after four weeks. The histomorphylogic structure of the liver tissues was observed under optical microscope; the levels of MDA and SOD in serum were determined by radioimmunoassay. Results  Compared with those in normal group, the collagen area in Masson staining and level of serum MDA were inreased obviously, and the level of serum SOD was dereased obviously in model group (P<0.05). Compared with those in model group, the collagen area and level of serum MDA were dereased obviously, and the level of serum SOD was inreased obviously in S�睯G group (P<0.05). Highly positive correlation between collagen area and level of serum MDA was observed (r=0.435, P<0.05), and significantly negative correlation between collagen area and level of serum SOD was found (r=-0.358, P<0.05). Conclusion  S�睯G has good anti�瞗ibrosis effects in rats with hepatic fibrosis, and its mechanism may be correlated with its anti�瞝ipid peroxidation.

　　KEY WORDS: hepatic fibrosis; Sanjia Yigan Granule; hepatic stellate cell; lipid peroxidation

　　脂質(zhì)過氧化(lipid peroxidation)在肝纖維化過程中發(fā)揮重要作用，脂質(zhì)過氧化的終產(chǎn)物丙二醛(malondialdehyde, MDA)可以激活靜息狀態(tài)的肝星型細(xì)胞(hepatic stellate cells, HSCs)，促使其合成與分泌大量的細(xì)胞外基質(zhì)(extracellular matrix, ECM)而最終導(dǎo)致肝纖維化的形成[1]。因此，可通過抗脂質(zhì)過氧化來達(dá)到抗肝纖維化的目的[2]。我院自制復(fù)方中藥三甲益肝沖劑(Sanjia�瞴igan Granule, S�睯G)具有活血化瘀、健脾益氣、舒肝理氣、軟堅(jiān)散結(jié)等作用，對治療肝炎后肝硬化和慢性肝炎已取得良好效果[3]。本研究以二甲基亞硝胺(dimethylnitrosamine, DMN)誘導(dǎo)大鼠肝纖維化，以復(fù)方丹參(compound Salviae miltiorrhizae, Sm)作為治療對照組，觀察三甲益肝沖劑對大鼠肝纖維化的作用，并進(jìn)一步探討其與抗脂質(zhì)過氧化的關(guān)系，以指導(dǎo)臨床用藥。

　　1  材料與方法

　　1.1  材料 專業(yè)論文發(fā)表網(wǎng)

　　雄性Spragne�睤awley(SD)大鼠，3月齡，40只，體重180-220g(新鄉(xiāng)醫(yī)學(xué)院實(shí)驗(yàn)動物中心提供)；DMN(日本化成公司)；三甲益肝沖劑(12g/袋，新鄉(xiāng)醫(yī)學(xué)院第一附屬醫(yī)院生產(chǎn)，產(chǎn)品批號071024，組方包括穿山甲、莪術(shù)、桃仁、丹參、郁金、赤芍、黃芪、白術(shù)、柴胡、內(nèi)金、大黃等十五味。以蒸餾水稀釋成0.34g/mL溶液)；復(fù)方丹參滴丸(天津天力士制藥股份有限公司生產(chǎn)，產(chǎn)品批號20070417，研細(xì)以蒸餾水稀釋成8.1g/L溶液)。超氧化物岐化酶(SOD)、MDA檢測試劑盒(南京建成生物工程研究所)。

　　1.2  動物分組及肝纖維化模型的建立

　　SD大鼠飼養(yǎng)籠飼養(yǎng)，自由飲水，標(biāo)準(zhǔn)飼料喂養(yǎng)。適應(yīng)環(huán)境1周后，隨機(jī)分4組：正常組、模型組、復(fù)方丹參組和三甲益肝沖劑組(n=10)。參考文獻(xiàn)[4]方法造模，將DMN 335μL加0.15mol/L的氯化鈉溶液中稀釋至66.96mL，制備腹腔注射液，每周連續(xù)3d，按1次/d腹腔注射上述DMN稀釋液2mL/kg(第1周為此劑量的2/3)，共4周。模型對照組造模同時予生理鹽水灌胃，復(fù)方丹參組造模同時給予復(fù)方丹參滴丸蒸餾水稀釋液灌胃(每日10mL/kg)，三甲益肝沖劑組造模同時給予三甲益肝沖劑蒸餾水稀釋液灌胃(每日10mL/kg)，正常對照組腹腔注射生理鹽水，用自來水灌胃，方法劑量同上，共4周。在造模過程中模型組死亡1只。

　　1.3  標(biāo)本留取

　　實(shí)驗(yàn)4周后，氯氨酮麻醉下，將動物取仰位固定于手術(shù)臺上，取左側(cè)第3-4肋間逐層切開暴露