积雪草苷对脊髓损伤大鼠神经元的保护作用的研究

目的探讨积雪草苷（asiaticoside,AS）对急性脊髓损伤（spinalcordinjury,SCI）大鼠神经元的保护作用及对诱导型一氧化氮合酶（induciblenitricoxidesynthase,iNOS）表达的影响。方法成年sD大鼠以Allen’s打击法制备脊髓损伤模型。分别给予不同剂量As治疗,术后24h、72h、7d、14d、28d采用BassoBeasttieandBresnahan（BBB）评分评价大鼠双后肢神经功能恢复情况,并通过HE和尼氏染色观察损伤脊髓组织病理变化,同步检测损伤部位脊髓的丙二醛（malondialdehyde,MDA）、超氧化物歧化酶（superoxidedismutase,SOD）值、iNOS蛋白表达及其活度值。结果与脊髓损伤模型组相比,低、中、高三剂量（15、30和45mg／kg）AS给药组在脊髓损伤后各时间点BBB评分分值明显提高（P〈0．05）,HE染色镜检脊髓组织病理损伤明显减轻,神经元尼氏体表达明显（P〈0．05）,MDA生成显著减少（P〈0．05）,SOD活性提高（P〈0．05）。各剂量AS均可下调SCI大鼠第7天受损脊髓组织iNOS蛋白表达和iNOS酶活度值。结论AS可减轻大鼠脊髓组织的损伤,保护脊髓神经元,此作用可能与抑制iNOS蛋白表达有关。     

积雪草苷对烫伤创面成纤维细胞及结缔组织生长因子表达的影响

目的探讨积雪草苷对烫伤创面成纤维细胞及结缔组织生长因子（CTGF）表达的影响。方法将30只SD大鼠按随机数字表法进行分组,选取6只大鼠不予烫伤,其余进行烫伤建模,采用积雪草苷和凡士林对烫伤创面组织进行同体对照研究,HE染色评估成纤维细胞变化,免疫组织化学法检测CTGF蛋白表达情况,并用透射电镜观察成纤维细胞微观结构改变。结果积雪草苷治疗组大鼠伤后第14天创面CTGF含量高于对照组（P〈0．05）,伤后第21天创面CTGF含量低于对照组（P〈0.05）。成纤维细胞HE染色的大体形态及电镜观察呈现双相动态变化。结论积雪草苷可能对成纤维细胞增殖及CTGF表达存在双相调节作用,对促进创面少瘢痕愈合有一定作用。     

Production of asiaticoside from centella (Centella asiatica L. Urban) cells in bioreactor

Objective

To investigate the effects of some culture conditions on production of asiaticoside from centella (Centella asiatica L. Urban) cells cultured in 5-L bioreactor.

Methods

The centell cell suspension culture was conducted in 5-L bioreactor to investigate the growth and asiaticoside accumulation under various conditions. Asiaticoside content was determined by HPLC analysis.

Results

The results showed that the cell growth and asiaticoside accumulation peaked after 24 d of culture at an agitation speed of 150 r/min and aeration rate of 2.5 L/min. The cell biomass reached a maximum value of 302.45 g fresh weight (31.45 g dry weight) and growth index of 3.03 with inoculum size of 100 g. However, asiaticoside content was the highest (60.08 mg/g dry weight) when culture was initiated with an inoculum size of 50 g.

Conclusions

The present study found the suitable conditions for growth of centella cells and their asiaticoside production in bioreactor.     

积雪苷霜软膏质量标准研究

目的:建立积雪苷霜软膏的质量控制方法。方法:用TLC法鉴别制剂中的羟基积雪草苷、积雪草苷,用HPLC法测定制剂中积雪草苷的含量。结果:鉴别方法专属性强,含量测定方法积雪草苷的平均回收率为97.97%,RSD为1.1%。结论:所建方法为制剂质量标准的制订提供了实验依据。     

积雪草苷对脑缺血再灌注大鼠Nrf-2/TXNIP/NLRP-3通路及小胶质细胞活化的影响

目的 观察积雪草苷(Asiaticoside,ASI)对脑缺血再灌注(Cerebral ischemia reperfusion,CIR)大鼠皮层中核因子相关因子-2(Nuclear factor-erythroid 2-related factor-2,Nrf-2)、氧还蛋白相互作用蛋白(Thiredoxin-int...     

积雪草甙诱导肿瘤细胞凋亡及增强长春新碱的抗肿瘤作用

背景与目的:中药积雪草的三萜类成分积雪草甙(asiaticoside,ATS)具有促进胶原蛋白合成作用,对创伤愈合有较好的疗效。近年有文献报道积雪草甙有一定的抗肿瘤活性。本文研究积雪草甙诱导肿瘤细胞凋亡及其与长春新碱(vincristine,VCR)的协同作用。方法:采用MTT法检测积雪草甙单用或合用VCR对KB、KBv200、MCF-7和MCF-7/ADM细胞的增殖抑制作用,流式细胞术分析KB细胞周期和凋亡率的变化,琼脂糖凝胶电泳、荧光显微镜和Westernblot方法观察KB细胞凋亡和细胞周期相关性质改变。结果:积雪草甙对KB、KBv200、MCF-7和MCF-7/ADM细胞的IC50分别为(1.11±0.13)、(1.82±0.08)、(1.58±0.15)mg/ml和(3.25±0.46)mg/ml,且KBv200和MCF-7/ADM细胞对积雪草甙表现出与相应的亲本细胞相近的药物敏感性。经积雪草甙处理后的KB细胞表现出凋亡细胞特征性的改变。积雪草甙与VCR合用对多种肿瘤细胞均有显著协同作用,合并用药组KB细胞凋亡率、Bcl-2蛋白磷酸化水平明显高于单用组,且仅有合用组检测到活化后的caspase-3蛋白。积雪草甙与VCR合用组KB细胞阻滞于S-G2/M期的比率和CyclinB1蛋白的表达水平高于单用组,P34cdc2蛋白表达略有降低。结论:积雪草甙可诱导肿瘤细胞凋亡并与长春新碱显示协同作用,有可能作为生化调节剂应用于肿瘤化     

优化积雪草苷非离子表面活性剂囊泡的处方和工艺

 目的 筛选积雪草苷非离子表面活性剂囊泡的最优处方和最佳制备工艺。方法 采用薄膜蒸发法制备积雪草苷囊泡,以包封率和载药量为指标对非离子型表面活性剂的种类和用量、投药量、胆固醇用量、孵化时间、孵化温度等处方因素和工艺条件进行单因素考察,正交设计筛选处方、优化制备工艺,并对积雪草苷囊泡的最优处方和工艺进行验证。结果 因素考察试验中,对积雪草苷囊泡包封率和载药量影响的顺序依次为:投药量>孵化温度>孵化时间。优化处方及工艺为:积雪草苷36 mg,孵化温度30 ℃,孵化时间30 min,其包封率和载药量分别为89.56%和25.26%,镜下观察成典型囊形。结论 采用薄膜蒸发法所制备的积雪草苷囊泡呈圆形,分散性好,囊泡的包封率和载药量均较高。     

积雪草甙对烧伤增生性瘢痕作用的实验研究

目的 研究积雪草甙对体外培养的成纤维细胞的胶原蛋白合成的影响,以及对裸鼠增生性瘢痕动物模型的作用,以探讨积雪草甙治疗增生性瘢痕的机制。方法采用光镜、电镜研究用药前后成纤维细胞的形态学的变化,以^3H脯氨酸掺入等方法检测成纤维细胞胶原蛋白的合成,通过建立裸鼠增生性瘢痕动物模型,观察积雪草甙在体局部注射对增生性瘢痕的作用。结果 积雪草甙能明显影响成纤维细胞的超微结构,抑制胶原蛋白的合成,并呈量效关系;在裸鼠体内局部注射积雪草甙可抑制瘢痕增生,而且大剂量使用未发现药物毒性作用。结论 积雪草甙治疗瘢痕增生的机理与其抑制成纤维细胞增殖和胶原蛋白合成有关。     

滞留时间法评价促透剂对积雪草苷体外经皮释药特性的影响

目的考察薄荷油、冰片和氮酮对积雪草苷体外经皮渗透的影响。方法使用改良Franz体外透皮装置,采用反相高效液相色谱法检测接收液中积雪草苷的浓度并计算药物的经皮累积渗透量,采用滞留时间法评价不同促透剂对积雪草苷的经皮释药特性的影响。结果氮酮对积雪草苷的经皮促透作用最好,24 h内累积经皮渗透量为7.120μg/cm2,滞留时间适中为0.366 h。1%冰片和1%的薄荷24 h内累积经皮渗透量分别为5.025μg/cm2和4.660μg/cm2,滞留时间分别为0.195 h和0.547 h。结论氮酮、冰片均能够提高积雪草苷的透皮效率,薄荷油对积雪草苷的促透作用较差。     

Comparative pharmacokinetics between madecassoside and asiaticoside presented in a standardised extract of Centella asiatica,ECa 233 and their respective pure compound given separately in rats

1.?ECa 233, the standardised extract of Centella asiatica, contains not less than 80% triterpenoid glycosides, in a madecassoside:asiaticoside ratio of 1.5 (±0.5):1.

2.?The pharmacokinetic comparison of madecassoside and asiaticoside was performed in rats following intravenous and oral administration of ECa 233, or an equivalent dose of the individual compounds. Blood, tissues, urine and faeces were collected after dosing to determine drug and metabolite levels using liquid chromatography–tandem mass spectrometry.

3.?Our study demonstrated that plasma levels of madecassoside, and to a lesser extent asiaticoside, were higher after administration of ECa 233 than the corresponding values for the pure compounds. There was a bidirectional interconversion between asiaticoside and madecassoside consistent with the increased exposure of madecassoside and asiaticoside in ECa 233.

4.?Both madecassoside and asiaticoside appeared to be widely distributed in several organs and metabolized extensively; following intravenous administration of either compound, approximately 80–90% of the dose was recovered as madecassic acid and asiatic acid in the faeces.