水飞蓟宾自微乳化胶囊剂的制备及大鼠体内药动学

目的 研制水飞蓟宾自微乳化胶囊剂并对其进行体内外评价.方法 通过正交设计和伪三元相图的绘制,对自微乳化系统中的油相、乳化剂及助乳化剂的组成、用量进行研究,筛选最佳处方组成和组成比例.以自制水飞蓟宾胶囊作对照,RP-HPLC测定大鼠灌胃后的血药浓度,用3p97计算药物动力学参数.结果 水飞蓟宾最佳自微乳化给药系统处方组成及其比例为水飞蓟宾-中链甘油三酯-Cremophor RH40-PEG400=3.85∶16.15∶60∶20,所形成的微乳的平均粒径为14.6 nm,在人工胃液和人工肠液中16 min内累积溶出百分率均超过95％,自制水飞蓟宾颗粒胶囊溶出很少.胶囊内容物水飞蓟宾预乳化浓缩液(SLB-PMC)和水飞蓟宾颗粒浓度-时间数据符合一级吸收单室模型,水飞蓟宾预乳化浓缩液的Cmax为0.70 μg/mL,而自制水飞蓟宾颗粒仅为0.104 μg/mL,水飞蓟宾预乳化浓缩液的AUC比水飞蓟宾颗粒提高了11.7倍,其相对生物利用度为1 265.22％.结论 将水飞蓟宾制成自微乳化胶囊能显著提高其体外溶出和体内吸收.     

水飞蓟宾对成年大鼠分离的心肌细胞质膜Ca2+通道 活性的影响

 目的：观察药物水飞蓟宾对分离的成年大鼠心肌细胞钙通道阻滞效应。方法：用成年大鼠分离的心肌细胞，以放射性同位素⁴⁵Ca²⁺作示踪，通过对经细胞膜Ca²⁺流入测定，研究药物水飞蓟宾对钙通道的抑制效应，并与钙通道阻滞剂维拉帕米对照。结果：维拉帕米组Ca²⁺流入的平均抑制效应为30%左右（P<0.001),水飞蓟宾组为36.50%左右(P<0.001)。结论：药物水飞蓟宾能显著阻滞分离的成年大鼠心肌细胞胞膜钙通道。     

Plasma lipoproteins in transport of silibinin,an antioxidant flavonolignan from Silybum marianum

To assess the role of plasma lipoproteins in the transport of silibinin, an antioxidant flavonolignan, (125)I-labelled silibinin ((125)I-SB) administered perorally to the rat was used. The plasma (125)I-SB derived radioactivity was distributed among plasma lipoproteins according to their lipophilicity (TAG-rich lipoproteins 30-40% > LDL 15% > HDL 5%), and in the fraction of d > 1.215 containing albumin and other proteins a minority amount of radioactivity was found. Administration of (125)I-SB in a complex with phosphatidylcholine resulted in proportionally higher radioactivities in all fractions as well as in tissues. Dietary olive oil had a slightly decreasing effect on plasma concentrations of silibinin measured by HPLC as well as on (125)I-SB derived radioactivity in plasma and liver. In the TAG-rich lipoprotein fraction and HDL no effects of olive oil on the levels of (125)I-SB derived radioactivities were observed, however, at a 30 min interval the levels of (125)I-SB derived radioactivity in LDL and the heart were significantly decreased in the olive oil group. These results suggest that (i) silibinin is not resorbed by the chylomicron pathway, and (ii) the endogenous lipoprotein pathway VLDL --> LDL may play a role in the transport of silibinin from the liver to the extrahepatic tissues concurrently facilitating the lipoprotein antioxidant influence of silibinin.     

Effects of Flavonoids on the Resistance of Microsomes to Lipid Peroxidation In Vitro and Ex Vivo

Incubation of rat liver microsomes with preparations of grape flavonoids, dihydroquercetin, and silibinin increased their resistance to lipid peroxidation induced by NADPH-Fe2+. This was manifested in less pronounced accumulation of lipid peroxidation products and changes in activity of microsomal enzymes induced by lipid peroxidation. In vitro antioxidant activity of grape flavonoids markedly surpassed that of dihydroquercetin and silibinin. Addition of flavonoids into fodder led to moderate, statistically significant, and similar increase in the resistance of rat liver microsomes to ex vivo induced lipid peroxidation.     

水林佳联合复方丹参滴丸治疗脂肪肝和肝纤维化22例疗效观察

目的:观察水林佳(水飞蓟宾磷脂复合物)联合复方丹参滴丸治疗脂肪肝和肝纤维化临床疗效。方法:43例脂肪肝患者随机分为2组,对照组21例给予一般的保肝、降脂治疗(甘利欣、护肝片、脂必妥、维生素C等);治疗组22例在对照组治疗的基础上加用水林佳胶囊及复方丹参滴丸口服。结果:治疗组总有效率95.5%,对照组66.7%,经统计学处理,两组有显著性差异。治疗后治疗组肝纤维化指标较对照组明显降低。结论:水林佳联合复方丹参滴丸对脂肪肝、肝纤维化以及伴有高血脂的患者有较好疗效。     

Silibinin Effect on Fas/FasL,HMGB1, and CD45 Expressions in a Rat Model Subjected to Liver Ischemia-Reperfusion Injury

ABSTRACT

Purpose: We investigated the hepatoprotective effect of Silibinin (SLB) to ischemia-reperfusion (I/R) rat model, by evaluating the histological expression of the tissue markers Fas/FasL, HMGB-1 and CD45, and SLB pharmacokinetics. Methods: Seventy-three Wistar-type male rats were randomized in 11 groups: Sham control group (open-close laparotomy); four I/R control groups (laparotomy, 45 min vascular occlusion, reperfusion, euthanasia after 60, 120, 180, and 240 min); four SLB (Si) groups (laparotomy, 45 min vascular occlusion, IV administration of SLB, reperfusion, euthanasia after 60, 120, 180, and 240 min); two SLB pharmacokinetics (PK) groups (IV administration of SLB, euthanasia after 45 and 240 min). Results: Fas/FasL increased with reperfusion time in I/R control groups and decreased in the Si groups, reaching, respectively, the highest and lowest values at 240 min of reperfusion (p <.0001). HMGB1 and CD45 increased with time in the I/R control groups up to 240 min and decreased in the Si groups, approaching zero expression after 180 and 60 min, respectively. Pharmacokinetic data showed higher liver accumulation and slower plasma elimination of SLB in ischemic animals. Conclusions: The hepatoprotective effect of SLB was demonstrated through the reduction of the expression of Fas/FasL, HMGB-1 and CD45 in liver tissue under I/R conditions, and in the pharmacokinetic study. The results document the efficacy of silibinin in the protection of the liver, and are particularly encouraging for its use in hepatic surgery.     

水飞蓟宾诱导人肝癌细胞HepG2死亡及机制研究

目的：探讨水飞蓟宾对人肝癌细胞HepG2的杀伤作用及其机制。方法不同浓度水飞蓟宾处理HepG2细胞,采用MTT法检测HepG2细胞增殖情况,流式细胞术检测水飞蓟宾诱导HepG2细胞的凋亡情况。利用ROS荧光探针二氢乙啶（DHE）检测水飞蓟宾是否诱导HepG2细胞ROS的产生,并用ROS抑制剂抗坏血酸预处理后检测水飞蓟宾对HepG2的杀伤活性及诱导凋亡的能力。结果水飞蓟宾可显著抑制HepG2细胞的增殖并诱导其发生凋亡。水飞蓟宾处理后HepG2细胞的ROS显著增高（P〈0.05,P〈0.01）,用抗坏血酸预处理后水飞蓟宾对HepG2的增殖抑制作用降低,并抑制水飞蓟宾诱导的凋亡。结论水飞蓟宾通过诱导ROS的产生引起人肝癌细胞发生凋亡。     

Silibinin-mediated metabolic reprogramming attenuates pancreatic cancer-induced cachexia and tumor growth

Pancreatic ductal adenocarcinoma (PDAC) is the fourth leading cause of cancer-related deaths in the US. Cancer-associated cachexia is present in up to 80% of PDAC patients and is associated with aggressive disease and poor prognosis. In the present studies we evaluated an anti-cancer natural product silibinin for its effectiveness in targeting pancreatic cancer aggressiveness and the cachectic properties of pancreatic cancer cells and tumors. Our results demonstrate that silibinin inhibits pancreatic cancer cell growth in a dose-dependent manner and reduces glycolytic activity of cancer cells. Our LC-MS/MS based metabolomics data demonstrates that silibinin treatment induces global metabolic reprogramming in pancreatic cancer cells. Silibinin treatment diminishes c-MYC expression, a key regulator of cancer metabolism. Furthermore, we observed reduced STAT3 signaling in silibinin-treated cancer cells. Overexpression of constitutively active STAT3 was sufficient to substantially revert the silibinin-induced downregulation of c-MYC and the metabolic phenotype. Our in vivo investigations demonstrate that silibinin reduces tumor growth and proliferation in an orthotopic mouse model of pancreatic cancer and prevents the loss of body weight and muscle. It also improves physical activity including grip strength and latency to fall in tumor-bearing mice. In conclusion, silibinin-induced metabolic reprogramming diminishes cell growth and cachectic properties of pancreatic cancer cells and animal models.     

High-dose silibinin rescue treatment for HCV-infected patients showing suboptimal virologic response to standard combination therapy

Incomplete suppression of hepatitis C virus (HCV) replication with persistence of minimal viremia (partial virologic response) leading to treatment failure can be observed in a significant proportion of HCV type 1-infected patients during antiviral therapy. Recently, high-dose intravenous silibinin has demonstrated strong antiviral activity against HCV. We were therefore interested in whether patients with partial virologic response can be rescued by the on-treatment addition of a short-term course of high-dose intravenous silibinin infusions. Twenty patients who failed to achieve a complete virologic response to different interferon-based regimens qualified for the rescue strategy and received 1400 mg/day silibinin infusions on two consecutive days. Complete viral suppression (below the limit of detection <6 IU/mL, TMA assay) could be induced in 13 of 20 patients within the first week after the short-term silibinin infusion, and all but one of them also remained HCV RNA negative during the subsequent follow-up period on continued peginterferon plus ribavirin treatment. In the remaining seven patients, no complete suppression could be achieved although four showed a significant HCV RNA reduction in response to silibinin. Silibinin infusions were generally well tolerated, and activation of abdominal peristalsis with nausea, diarrhoea and vomiting were the most prominent side effects. Of the twelve patients who exhibited a durable response to peginterferon and ribavirin treatment, three achieved an SVR, two achieved a week 12 SVR and four suffered a viral relapse. Three patients could not complete the assigned antiviral treatment with peginterferon alpha and ribavirin for nonvirological reasons. Short-term administration of high-dose intravenous silibinin might be an interesting approach to rescue patients with ongoing minimal residual viremia while on interferon-based therapy. These preliminary findings may stimulate further studies to evaluate more refined therapeutic strategies.     

拉米夫定联合水飞蓟宾治疗对E抗原阳性的慢性乙型肝炎患者相关血液指标的影响

目的：应用拉米夫定联合水飞蓟宾治疗 E 抗原阳性的慢性乙型肝炎,通过治疗前后的相关血液指标变化判断疗效,为个体化治疗提供依据。方法选取 E 抗原阳性慢性乙型肝炎患者226例,按肝功能损伤程度分为轻度组55例、中度组94例和重度组77例,均给予口服拉米夫定100 mg,每日1次及服用水飞蓟宾70 mg,每日3次,疗程6个月。分别对治疗前后的相关血液指标进行分析。结果治疗6个月后,轻度组及中度组患者血清 ALT 复常率、E 抗原阴转率及 HBV-DNA 阴转率有不同程度的改善,而重度组上述指标则均显著提高（P ＜0．05）。结论拉米夫定联合水飞蓟宾治疗更能有效抑制 HBV-DNA 的复制,明显促进血清 ALT 复常率及提高 E 抗原阴转率,尤其是肝功能损伤程度重者;在一定程度上提高治疗慢性乙型肝炎的疗效。