蓝莓对肝纤维化大鼠血红素加氧酶-1表达的影响

Effects of blueberry on the expression patterns of heme oxygcnase-1 in rats with hepatic fibrosis

目的 观察蓝莓对大鼠肝纤维化的预防作用及对血红素加氧酶-1（HO-1）表达的影响.方法 45只SD大鼠随机分为生理盐水对照组（A组）、模型组（B组）、蓝莓汁预防组（C组）、丹芍化纤胶囊预防组（D组）及蓝莓汁加丹芍化纤胶囊预防组（E组）,每组9只.CCl4复合因素制备大鼠肝纤维化模型,各预防组给予蓝莓汁或（和）丹芍化纤胶囊灌胃,共饲养8周.测定各组大鼠血清ALT水平及肝组织匀浆中超氧化物歧化酶（SOD）和丙二醛（MDA）的表达,并进行肝组织病理学检查.采用实时荧光定量聚合酶链反应、免疫组织化学法和Western blot分析大鼠肝组织HO 1的mRNA及蛋白质表达.多组间数据比较用单因素方差分析,等级资料用秩和检验.结果 B组大鼠血清ALT水平高于A组[（203.25±31.62）U/L比（57.25±6.88）U/L,F=92.498,P＜0.05）],各预防组较B组明显减少[C、D和E组分别为（149.44±16.51）U/L、（136.88±10.07）U/L和（127.38±11.03）U/L,F=92.498,P＜0.05）].C、D、E组大鼠肝组织匀浆SOD水平分别为（1.36±0.09）U/mg、（1.42±0.13）U/mg、（1.50±0.15）U/mg,高于B组的（1.08±0.19）U/mg（F=13.671,P＜0.05）;MDA水平分别为（0.294±0.026）nmol/mg、（0.285±0.025）nmol/mg、（0.284±0.028）nmol/mg,低于B组的（0.335±0.056）nmol/mg（F=20.809,P＜0.05）.各预防组肝纤维化程度较B组也有所减轻（x2-24.956,P＜0.05）.B组大鼠肝组织HO-1的mRNA和蛋白质表达高于A组（F值分别为4.549和22.926,P值均＜0.05）,各预防组大鼠肝组织HO-1的mRNA和蛋白质表达均高于B组,但差异无统计学意义（P＞0.05）.结论 蓝莓对CCl4所致大鼠肝纤维化有一定的预防作用;在慢性肝损伤时,蓝莓对HO-1的表达无明显影响.

Objective To study the protective effect of Blueberry against rat hepatic fibrosis and the effect of Blueberry on HO-1 expression patterns. Methods Atotal of 45 SD rats were randomly divided into five groups namely control group （group A）, model group （group B）, blueberry group （group C）, Dan-shaohua-xian （DSHX） capsule group （group D） and blueberry ＋Dan-shao-hua-xian group （group E）. Fibrous liver models in rats were induced by subcutaneous injection of CC14 and high-lipid/low-protein diet for 8 weeks except the control group which accepted saline alone. The level of alanine aminotransferase （ALT） in serum was examined. The activities of superoxide dismutase （SOD） and malondialdehyde （MDA） in liver homogenates were determined. by the xanthine oxidase method and the thiobarbituric acid method. The pathology of hepatic fibrosis was evaluated by hematoxylin and eosin （H＆E） staining. The Expression of HO-1 was detected by real-time RT-PCR, immunohistochemical techniques and western blotting. Results Serum ALT levels in every prevention group was lower than the group B [（149.44 ± 16.51）, （136.88 ± 10.07）, （127.38 ± 11.03）vs （203.25 ± 31.62） U/L, F = 92.498, P 〈 0.05], the SOD of liver homogenate in prevention group was significantly higher and the MDA was lower compared with the group B [SOD： （1.36 ± 0.09）, （1.42 ±0.13）, （1.50 ± 0.15） vs （1.08 ± 0.19） U/mg, F= 13.671, P 〈 0.05; MDA： （0.294 ± 0.026）, （0.285 ±0.025）, （0.284 ± 0.028） vs （0.335 ± 0.056） nmol/mg, F = 20.809, P 〈 0.05]. The pathological stages of hepatic fibrosis were all significantly reduced in prevention group （ x 2 = 24.956, P 〈 0.05）. Compared with group A, the mRNA and protein expressions of HO-1 were elevated （F = 4.549, 22.926, P 〈 0.05） in group B and increased in group C-E, but there is no significant difference existed. Conclusion Blueberry may have preventive and protective effects on CCl4-induced hepatic fibrosis by reducing lipid peroxidation. However,these effects may not be related to the activation of HO-1 during long-term of CCl4.