Effects of Aqueous Extract of Plumbago zeylanica L. in the Reversal of DMN-induced Hepatic Fibrosis in Rat

[ Objectives ] This study was conducted to evaluate the action of the aqueous extract of a medicinal plant Plumbago zeylanica L. in the reversal of dime- thylnitrosamine （DMN） -induced hepatic fibrosis in rats, and to provide a scientific basis for the utilization of P. zeylanica in treatment of hepatic fibrosis. [Meth- ods ] Hepatic fibrosis in rats was induced by intraperitoneal injection of DMN. The rats were then given the aqueous extract of P. zeylanica at high, medium or low concentration by garage for five weeks. Serum level of alanine aminotransferase （ALT） was determined by lactate dehydrogenase （LDH）-release assay, and serum level of aspartate transaminase （AST） was measured by UV-malate dehydrogenase （MDH） assay. Serum levels of total bilirubin （TBIL）, direct bilirubin （DBIL） and indirect bilirubin （1BIL） were measured by vanadate oxidation assay. Four indices of hepatic fibrosis （hyaluronic acid, laminin, procollagen type III and colla- gen type IV） were determined by radioimmunoassay （RIA） assay. Morphological damage of liver tissue was observed by hematoxylin-eosin staining （H＆E stai- ning）. Immunohistochemical staining was performed to determine the location and area of deposited collagen type I, collagen type III and ct-smeoth muscle actin （a-SMA） in liver tissue. [ Results] Compared with the negative control （rats with diseased fiver and untreated with P. zeylanica aqueous extract）, the serum lev- els of ALT, AST, TBIL, DBIL and IBIL were significantly decreased by P. zeylanica aqueous extract; the levels of the four serum indices of hepatic fibrosis were also obviously reduced. H＆E staining showed that hepatic fibrosis in rats was obviously inhibited or even reversed by P. zeylanica aqueous extract. Immunohisto- chemical staining proved that the aqueous extract of P. zeylanica significantly reduced the area and coloration of collagen type I, collagen type III and ct-SMA in rat liver. [ Conclusions] The aqueous extract of P. zeylanica has a definite effect in reversal of DMN-indueed hepatic fibrosis in rat by promoting the recovery of liver function, reversal of histopathological changes and reducing fibrotic collagen.

响应面法优化兰雪醌超声辅助提取工艺的研究

目的优化白花丹中兰雪醌超声辅助提取的最佳工艺。方法采用紫外分光光度法分析兰雪醌含量。采用单因素试验考察超声时间、提取温度和料液比对兰雪醌得率的影响,并在单因素试验的基础上使用Box-Behnken响应面法优化得到最佳工艺。结果该模型的调整决定系数(R_(Adj)^(2))为0.8394,回归模型的概率值(P<0.005)显示出较高的显著性。通过Box-Behnke响应面法得到超声提取兰雪醌的最佳工艺参数为超声时间41 min,提取温度42℃,料液比1∶23(g/mL),预测兰雪醌得率为1.823%。在最佳条件下进行了5次验证试验,得到兰雪醌平均得率为1.8941%,与预测值基本吻合。结论利用超声辅助提取方法提取白花丹中兰雪醌的工艺稳定可靠。

不同溶剂索氏提取的白花丹叶中白花丹醌的含量测定

[目的]测定不同方法提取的白花丹鲜叶中白花丹醌的含量并加以比较,优选出最佳提取溶剂。[方法]采用索氏提取法,选用不同的溶剂提取白花丹鲜叶中的白花丹醌,并通过HPLC测定其含量,从而确定最佳提取溶剂。[结果]测得用氯仿、石油醚(II)、浓度95%乙醇、无水甲醇提取白花丹醌的含量依次为0.511%、0.341%、0.233%和0.191%;用氯仿提取的白花丹醌含量(提取率)最高。[结论]不同方法提取的白花丹鲜叶中白花丹醌的含量差异明显。

微波辅助提取白花丹中黄酮的最佳工艺研究

目的:优选白花丹中总黄酮的提取工艺。方法:利用正交实验优选总黄酮最佳提取工艺,考察乙醇浓度、料液比、微波辐射功率、微波辐射时间对总黄酮提取率的影响。结果:总黄酮的最佳工艺条件为乙醇浓度60%、料液比1∶25、微波辐射功率400 W、微波辐射时间60 s。结论:微波提取法可用于白花丹总黄酮的提取,其中乙醇浓度对总黄酮的提取影响最大。

白花丹提取物对高糖诱导的肾小球系膜细胞分泌炎症因子及细胞外基质的影响

目的探讨白花丹提取物对高糖诱导的肾小球系膜细胞分泌炎症因子及细胞外基质的影响。方法体外培养大鼠肾小球系膜细胞,随机分为对照组,模型组,白花丹提取物低剂量(1 mg/L)、中剂量(2 mg/L)、高剂量(4 mg/L)组,甘露醇(24.5 mmol/L)组,收集各组细胞及细胞培养基,以酶联免疫吸附法检测各组细胞上清液中肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白细胞介素6(interleukin-6,IL-6)、转化生长因子β1(transforming growth factor-β1,TGF-β1)水平;检测各组细胞超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)水平;以实时荧光定量法检测各组细胞miR-155、SOCS1 mRNA水平;采用蛋白免疫印迹法检测各组细胞纤维连结蛋白(fibronectin,FN)、Ⅳ型胶原蛋白(collagen IV,Col IV)、细胞因子信号传导抑制蛋白1(cytokine signal transduction inhibitor 1,SOCS1)蛋白表达。结果与对照组相比,模型组、甘露醇组细胞上清液中TNF-α、IL-6及TGF-β1水平升高,细胞MDA、miR-155、FN及Col IV表达升高(P<0.05),细胞SOD水平、SOCS1 mRNA及蛋白表达明显降低(P<0.05)。与模型组相比,白花丹提取物低、中、高剂量组细胞上清液中TNF-α、IL-6及TGF-β1水平降低,细胞MDA、miR-155、FN及Col IV表达降低(P<0.05),细胞SOD水平、SOCS1 mRNA及蛋白表达升高(P<0.05),且各项变化均对白花丹提取物呈剂量依赖性(P<0.05)。结论白花丹提取物可抑制高糖诱导的肾小球系膜细胞分泌炎症因子及细胞外基质,下调miR-155表达、上调SOCS1表达可能是其作用机制。

白花丹炮制前后长期毒性的研究

目的观察连续灌胃白花丹水煎液和白花丹炮制品水煎液导致大鼠慢性毒性的损伤表现、程度及可逆性。方法140只SD大鼠随机分为空白对照组、白花丹水煎液高、中、低剂量组,白花丹炮制品水煎液高、中、低剂量组,大鼠连续灌胃12周,给药期间每天观察动物的一般情况,停药24 h后,分别检测血常规、血生化,测定主要脏器指数。结果连续灌胃白花丹水煎液和白花丹炮制品水煎液12周,对大鼠体重的增加没明显影响;白花丹水煎液的血常规RBC、PLT、MCH值出现明显升高;白花丹水煎液血清ALB、A/G、AST、CREA、CK含量显著下降;白花丹炮制品水煎液血清ALP、GLU有显著升高;白花丹炮制前肾脏指数和肾上腺指数出现显著性升高,炮制后无显著性差异。结论白花丹水煎液长期给药对大鼠的血常规和血清有一定影响,经酒制炮制后,影响减少。