Zhou Xian, Dai Li-li, Jia Li-ping, Zheng Yuan-yi, Wang Wen-bing
Department of Gastroenterology, Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China.
Zhonghua Gan Zang Bing Za Zhi. 2005 Aug;13(8):575-8.
To investigate the effect of Astragalus Injection solution on rat hepatic stellate cells (HSC) and hepatic fibrosis.
HSCs of rats were incubated with various concentrations of Astragalus Injection solution (0 mg/ml, 25 mg/ml, 50 mg/ml, 100 mg/ml, 200 mg/ml, 400 mg/ml) for 24, 48 and 72 hours. Cell proliferation was detected with 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphennyltetrazolium bromide (MTT) colorimetric assay. Cell cycle was detected with flow cytometry. Cell apoptosis was detected with acridine orange/ethidium bromide (AO/EB) fluorescent staining and flow cytometry. In vivo, rats were randomly allocated into a normal control group, a model control group and an Astragalus Injection group. Astragalus Injection (800 mg.kg-1.d-1) was administered to rats of the Astragalus Injection group. Rats of the model control group received saline. Serum concentrations of hyaluronic acid (HA) and laminin (LN), hepatic tissue activity of superoxide dismutase (SOD), and hepatic tissue contents of malondialdehyde (MDA) were measured in these groups at 8 weeks. Hepatic tissue expression of LN was assessed by using immunohistochemistry. The pathological changes of hepatic tissues were examined by hematoxylin-eosin (HE) and van Gieson (VG) staining of their histological slides.
In vitro, compared with the 0 mg/ml group, the proliferation of HSCs in other concentration groups was significantly inhibited by Astragalus Injection solution in a dose and time dependent manner, the cell proliferation cycle of HSCs was blocked in the G2-M phase, there was no apoptosis of HSCs in AO/EB fluorescent staining and flow cytometry. In vivo, compared with rats of the model control group, the rats of the Astragalus Injection solution treated group had remarkably decreased serum HA and LN levels (114.3+/-25.6) microg/L vs (85.6+/-37.3) microg/L and (78.8+/-11.7) microg/L vs (66.8+/-17.6) microg/L, P < 0.05, and liver MDA level (3.7+/-0.4) micromol/g protein vs (2.4+/-0.2) micromol/g protein, P < 0.01, but had increased activity of liver SOD (49.6+/-5.7) NU/mg protein vs (75.9+/-5.9) NU/mg protein, P < 0.01. Microscopic studies revealed that the livers of rats receiving Astragalus Injection solution showed decreases in fibrosis and in expression of LN.
Astragalus Injection solution has an inhibitive effect on experimental hepatic fibrogenesis. The mechanisms of its effects might possibly be associated with its antioxidant activity, expression of decreasing LN and its inhibition of HSCs proliferation.
探讨黄芪注射液对大鼠肝星状细胞（HSC）及肝纤维化的影响。
将大鼠HSC与不同浓度的黄芪注射液（0mg/ml、25mg/ml、50mg/ml、100mg/ml、200mg/ml、400mg/ml）孵育24、48和72小时。采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）比色法检测细胞增殖。用流式细胞术检测细胞周期。采用吖啶橙/溴化乙锭（AO/EB）荧光染色和流式细胞术检测细胞凋亡。在体内，将大鼠随机分为正常对照组、模型对照组和黄芪注射液组。黄芪注射液组大鼠给予黄芪注射液（800mg·kg-1·d-1）。模型对照组大鼠给予生理盐水。8周时检测这些组大鼠血清透明质酸（HA）和层粘连蛋白（LN）浓度、肝组织超氧化物歧化酶（SOD）活性以及肝组织丙二醛（MDA）含量。采用免疫组织化学法评估肝组织LN表达。通过苏木精-伊红（HE）和维多利亚蓝（VG）染色观察肝组织病理变化。
体外实验中，与0mg/ml组相比，黄芪注射液在其他浓度组均以剂量和时间依赖性方式显著抑制HSC增殖，HSC细胞增殖周期阻滞于G2-M期，AO/EB荧光染色和流式细胞术检测未发现HSC凋亡。体内实验中，与模型对照组大鼠相比，黄芪注射液治疗组大鼠血清HA和LN水平显著降低（114.3±25.6）μg/L对（85.6±37.3）μg/L和（78.8±11.7）μg/L对（66.8±17.6）μg/L，P<0.05，肝组织MDA水平（3.7±0.4）μmol/g蛋白对（2.4±0.2）μmol/g蛋白，P<0.01，但肝组织SOD活性升高（49.6±5.7）NU/mg蛋白对（75.9±5.9）NU/mg蛋白，P<0.01。显微镜检查显示，接受黄芪注射液的大鼠肝脏纤维化及LN表达降低。
黄芪注射液对实验性肝纤维化有抑制作用。其作用机制可能与其抗氧化活性、降低LN表达及抑制HSC增殖有关。
