复方鳖甲软肝片减方对CCl_4诱导大鼠肝纤维化的影响及机制研究  

Effect of modified formula Compound Biejia Ruangan Pills on hepatic fibrosis in rats induced by CCl_4 and its mechanism

作　　者：邓莉[1,2] 申宝德 刘园[2] 刘肖[2] 连王权[2] 李寅超 袁海龙[2] DENG Li1, 2, SHEN Bao-de2, LIU Yuan2, LIU Xiao2, LIAN Wang-quart2, LI Yin-chao1, YUAN Hai-long2(1. Zhengzhou University, Zhengzhou 450001, China 2. Air Force General Hospital, Beijing 100036, Chin)

机构地区：[1]郑州大学,河南郑州450001 [2]中国人民解放军空军总医院,北京100036

出　　处：《中草药》2018年第6期1371-1378,共8页Chinese Traditional and Herbal Drugs

基　　金：内蒙古科技计划重点项目(2015ZY0024);中国肝炎防治基金会-王宝恩肝纤维化研究基金资助课题(WBE20170066)

摘　　要：目的探讨复方鳖甲软肝片(CBRP)减去处方中紫河车后(即复方鳖甲软肝片减方,MF-CBRP)对肝纤维化大鼠的保护作用及其机制。方法将SD大鼠随机分为对照组,模型组,阳性对照组(秋水仙碱,0.1 mg/kg),CBRP组(0.594 7g/kg)和MF-CBRP高、低剂量(1.061 0、0.530 5 g/kg)组。除对照组外,其余各组大鼠sc CCl_4大豆油溶液制备大鼠肝纤维化模型,连续6周。造模结束后,各给药组ig对应剂量的药物治疗6周。给药结束后,检测大鼠血清生化指标、脂质过氧化指标以及纤维化因子、羟脯氨酸(Hyp)、血小板衍生因子-α(PDGF-α)和结缔组织生长因子(CTGF)量的变化;观察肝组织HE染色和Masson染色的变化;应用免疫组化和RT-PCR法检测大鼠肝组织中α-平滑肌肌动蛋白(α-SMA)和转化生长因子-β1(TGF-β1)、基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶抑制因子-1(TIMP-1)m RNA表达水平。结果与模型组比较,MF-CBRP可以使大鼠血清中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、总胆红素(T-Bil)和碱性磷酸酶(ALP)水平降低(P<0.01),AST/ALT、总蛋白(TP)和白蛋白(ALB)水平显著增高(P<0.05、0.01),使超氧化物歧化酶(SOD)和还原型谷胱甘肽(GSH)量升高(P<0.05)、丙二醛(MDA)量降低(P<0.01),抑制肝组织中Hyp、透明质酸(HA)、层黏连蛋白(LN)、III型前胶原(PC-III)和IV型胶原(IV-C)的表达(P<0.05、0.01),使肝组织中PDGF-α和CTGF量降低(P<0.01),抑制MMP-2和TIMP-1 m RNA表达(P<0.01),降低α-SMA免疫组化的阳性表达面积(P<0.01),改善肝组织病理学结构病变。与CBRP比较,MF-CBRP抗肝纤维化效果轻微减弱。结论 MF-CBRP通过减轻氧化胁迫、抑制胶原纤维增生、逆转肝星状细胞激活和抑制肝生长因子表达来抗大鼠肝纤维化,但其抗纤维化活性稍弱于CBRP。Objective To explore the possible effects and mechanism of modified Compound Biejia Ruangan Pills（MF-CBRP） on liver fibrosis induced by CCl_4 in rats. Methods SD rats were randomly divided into control group, model group, positive control（colchicine, 0.1 mg/kg） group, CBRP group（0.594 7 g/kg）, MF-CBRP high-and low-dose（1.061 0 and 0.530 5 g/kg） groups. The rat model with liver fibrosis was established by sc injection CCl_4 solution（dissolved in soybean oil）, twice a week for six weeks except control group. The rats in the treatment groups were administered six weeks after the model establishment. At the end of the administration, the contents of serum biochemical, hydroxyproline（Hyp）, lipid peroxidation, fibrosis factor, platelet-derived factor-α（PDGF-α）, and connective tissue growth factor（CTGF） were measured and the pathological changes of liver tissue were examined by HE and Masson. The level of α-SMA and the expression of transforming growth factor-β1（TGF-β1）, matrix metalloproteinase-2（MMP-2） and tissue inhibitor of metalloproteinase-1（TIMP-1） were detected by immunohistochemistry and RT-PCR. Results The results showed that MF-CBRP improved the liver function significantly through reducing the level of ALT, AST, T-Bil, ALP, Hyp, HA, LN, PC-III, IV-C, and MDA（P〈0.05, 0.01）, and increasing the content of S/L, TP, ALB, SOD, and GSH（P〈0.05, 0.01）. MFCBRP also reduced the expression of PDGF-α, CTGF, α-SMA, MMP-2, and TIMP-1 m RNA（P〈0.01） and improved the pathological changes of liver histopathology. Compared with CBRP, MF-CBRP anti-hepatic fibrosis effect was slightly weakened. ConclusionThe results suggested that MF-CBRP may against hepatic fibrosis by reducing oxidative stress, inhibiting collagen fibrillation, reversing hepatic stellate cell activation, and inhibiting the expression of hepatic growth factor. However its anti-fibrosis activity and mechanism was weaker than the original CBRP in a certain extent.

关 键 词：复方鳖甲软肝片减方 紫河车 肝纤维化 转化生长因子-β1 基质金属蛋白酶-2 基质金属蛋白酶抑制因子-1 

分 类 号：R285.5[医药卫生—中药学]