Microtox技术应用于生脉注射液综合毒性检测

目的：探索-Microtox技术应用于生脉注射液综合毒性检测。方法：以费氏弧菌为测试菌种,通过方法学考察确定最优检测体系及方法学可靠性;在最优检测体系条件下,首次以费氏弧菌对不同生产厂家所生产的生脉注射液进行发光菌综合毒性检测。结果：2mL反应体系下,最优复苏液体积0.9mL/支菌,每个待测样品加入最优菌液体积50μL,最优检测时间10min,最优pH范围5-10,且10min发光强度以80-120万为宜;重复性试验、中间精密度试验的相对标准偏差均<15%;不同生产厂家A、B、C成品的EC₅₀平均值分别为22.10%、34.10%、46.04%,具有极显著性差异（P<0.01）。结论：生脉注射液对费氏弧菌的毒性存在显著的浓度-效应关系,且不同生产厂家之间成品EC₅₀值具有显著性差异,提示我们生脉注射液成品生物学检测标准存在进一步提升的空间,应用Microtox技术检测生脉射液综合毒性并用于控制不同厂家成品质量波动具有很好的应用前景。     

生脉注射液不良反应分析

生脉注射液是根据金元时期名医李东垣创立的著名古方生脉散(人参、麦冬、五味子)经科学方法提取精制而成的注射液,被国家中医药管理局指定为中医急诊科(室)必备中成药之一,具有益气固脱、养阳生津、生脉之功,广泛应用于休克、冠心病、心衰等循环系统疾病[1].随着临床应用的扩展,其不良反应有所报道,应引起重视.     

生脉注射液预防阿霉素急性心脏毒性40例疗效观察

目的观察生脉注射液对阿霉素急性心脏毒性的预防效果。方法将40例肿瘤患者随机分为治疗组与对照组各20例,均予阿霉素为主联合化疗,治疗组加用生脉注射液,对照组口服维生素C、维生素E、辅酶Q10;比较两组用药后不同时段的主要症状、心电图、心肌肌钙蛋白I改善情况,并进行心脏损伤分级评价。结果两组急性心脏毒性反应和CTnI损伤情况相近。结论生脉注射液预防阿霉素所致急性心脏毒性效果与传统预防药物相似。     

生脉（及参脉）注射液的不良反应

生脉注射液根据著名古方生脉散（人参、麦冬、五味子）经科学方法精制而成的注射液,参脉注射液是在生脉注射液基础上,由人参、麦冬提取加工而成,被国家中医药管理局指定为中医急诊科（室）必备中成药之一,广泛应用于休克、冠心病、心衰等循环系统疾病。随着临床应用的...     

生脉及丹参注射液联用预防抗肿瘤抗生素化疗心脏毒性反应35例

目的:观察生脉注射液、丹参注射液对蒽环类相关性心脏毒性的保护作用。方法:69例恶性肿瘤患者随机分成治疗组及对照组。治疗组35例,在以蒽环类抗肿瘤抗生素为主的一线方案化疗前3 d给予生脉注射液60 ml及丹参注射液30 ml分别加入5%葡萄糖注射液150 ml中静脉注射,日1次,两周1疗程;对照组34例,于化疗前3 d开始服用辅酶Q10每次20 mg,每日3次,两周一疗程。治疗前检查12导联心电图排除基础心脏病、心绞痛、陈旧性心肌梗死,治疗后复查。结果:两组心电图改变有显著性差异(P<0.05)。结论:生脉及丹参注射液可预防和减轻蒽环类引起的心脏毒性。     

生脉注射液防治表柔比星心肌毒性的临床观察

目的研究生脉注射液防治表柔比星心肌毒性的疗效。方法治疗组与对照组均予CAP方案辅助化疗,治疗组于化疗开始前2 d予生脉注射液60 mL每日1次静点,连用15 d。结果治疗组心电图异常比率为2%,对照组为20%,2组比较有显著性差异。治疗组与对照组左室射血分数比较有显著性差异(P<0.05),生脉注射液减轻了表柔比星对心脏EF的影响。结论生脉注射液可防治表柔比星的心肌毒性,为临床上适当加大表柔比星的化疗剂量,提高疗效提供了可能。     

生脉注射液药效学研究进展

生脉注射液为《内外伤辨感论》中生脉散的注射剂，古人对生脉散有“人有将死脉欲绝者，服此能复生之”的评价。现代临床上补广泛用于冠心病、心衰、休克、病毒性心肌炎、急性心肌梗死、肺心病、脑血管病等的治疗，具有广范的临床使用价值。现将其药效学研究进展综述如下。     

生脉注射液及参麦注射液的临床应用与研究

生脉注射液源于著名方剂生脉散,麦门冬加五味子、人参二味,为生脉散。生脉注射液由红参、麦冬、五味子三味药组成。人参甘温,大补元气,尤长于补脾肺之气,麦冬性味甘苦微寒,入肺心胃经,五味子性味酸温,入肺肾二经,五味子与麦冬配伍,酸甘合化,以滋耗伤之津液,三药合用,一补一清一敛,共奏益气生津,养阴敛汗之功。生脉注射液中红参、麦冬、五味子三药比例为1∶3.12∶1.56。重用麦冬,养阴为主,补以益气。运用生脉注射液必须注意审邪,辨证虚实,外邪已尽,仅见气津耗伤者,方可用。参麦注射液仅以红参、麦冬二味,较生脉针少五味子一味。既无金水相生…     

生脉注射液防治阿霉素所致心脏毒性临床观察

化疗是治疗恶性肿瘤主要方法之一,阿霉素(ADM)是目前使用量广泛,疗效较佳的化疗药物,而由ADM所引起的心脏毒性屡见不鲜。我科于1996年6月～1999年10月应用生脉注射液防治ADM所致的心脏毒性取得较好效果,现报道如下。     

生脉注射液辅助治疗非小细胞肺癌临床观察

我院肿瘤科于2005年1月-2007年12月采用生脉注射液与化疗联用治疗中晚期非小细胞肺癌(NSCLC)32例,并与同期单纯化疗的34例患者进行对照,取得满意疗效,现报道如下.     

Two xanthones from Swertia punicea with hepatoprotective activities in vitro and in vivo

Ethnopharmacological relevance

Swertia punicea Hemsl. (Gentianaceae) is more commonly known as “Ganyan-cao” and used mainly as a traditional Chinese folk medicine for the treatment of acute bilious hepatitis, cholecystitis, fever, intoxification and jaundice.

Materials and methods

The active hepatoprotective constituents of Swertia punicea were purified using various column chromatography techniques. The structures of two isolated compounds were determined on the basis of spectroscopic data interpretation such as NMR analysis. The hepatoprotective activities of isolated compounds were evaluated by using hepatotoxicity in vitro and dimethylnitrosamine-induced rat hepatic fibrosis in vivo, respectively.

Results

Two xanthones, 1, 7-dihydroxy-3, 4, 8-trimethoxyxanthone (1) and bellidifolin (2) were isolated from the stems of Swertia punicea. The compounds 1 and 2 exhibited notable hepatoprotective activities against carbon tetrachloride (CCl₄) -induced HepG2 cell damage, and effectively alleviated the levels of aspartate transaminase (AST), lactate dehydrogenase (LDH), superoxide dismutase (SOD) and malonic dialdehyde (MDA) induced by CCl₄ in a concentration-dependent manner. Co-treatment with compound 2 significantly increased the cell viability compared with N-acetyl-p-aminophenol (APAP) treatment. Compound 2 also alleviated APAP-induced hepatotoxicity by increasing glutathione (GSH) content and decreasing hydroxyl free radical (·OH) levels and reactive oxygen specises (ROS) production. In addition, the protective effect of compound 1 significantly alleviated DMN-induced liver inflammation and fibrosis. Oral administration of compound 1 recovered the reduction of albumin (ALB) and reversed the elevation of serum alanine transaminase (ALT), AST and total bilirubin (TBIL) in dimethylnitrosamine (DMN)-induced fibrotic rats. Severe oxidative stress induced in fibrotic rats was evidenced by a 1.5-fold elevation in MDA and a fall in the SOD activity, and treatment with compound 1 protected against these adverse effects. Recovery of rat liver tissue against DMN-induced hepatocellular necrosis, inflammatory changes and hepatic fibrosis by compound 1 is also confirmed by H&E and Masson stained histopathological evaluation of liver tissue.

Conclusion

Two xanthones from Swertia punicea exhibited hepatoprotective activities in vitro (compounds 1 and 2) and in vivo (compound 1), respectively.     

Kaempferitrin induces immunostimulatory effects in vitro

Ethnopharmacological relevance

Justicia spicigera is a plant used as immunostimulatory in Mexican traditional medicine. Recently, we showed that Justicia spicigera extracts exerted immunostimulatory effects and the major component of this extract was kaempferitrin (KM). This work shows a correlation between the medical traditional use of Justicia spicigera and kaempferitrin, its active compound.

Materials and methods

The in vitro immunostimulatory effects of KM were evaluated on the proliferation of murine splenocytes and macrophages, and human peripheral blood mononuclear cells (PBMC). The effects of KM on NO production, lysosomal enzyme activity and neutral red uptake were assayed in murine macrophages RAW 264.7. The effects of KM on the NK cell activity were also assayed.

Results

KM at 25 μM, the highest concentration tested, increased the proliferation of murine macrophages (23%) and splenocytes (17%), and human PBMC (24%) in the absence of lipopolysaccharides (LPS), compared to untreated cells. KM also stimulated the pinocytosis (25%) and lysosomal enzyme activity (57%) in murine macrophages with a similar potency than LPS 1 μg/ml. In addition, KM induced the NK cell activity (11%).

Conclusion

KM exerts immunostimulatory effects on immune responses mediated by splenocytes, macrophages, PBMC and NK cells.     

Evaluating the anti-inflammatory potential of Tectaria cicutaria L. rhizome extract in vitro as well as in vivo

Ethnopharmacological relevance

The rhizome of Tectaria cicutaria has been used in the folklore system of Indian traditional medicine (Ayurveda) for the treatment of various disorders such as rheumatic pain, chest complaints, burns, sprain, poisonous bites, tonsilitis, toothache, gum complaints, cuts and wounds. The present work has for the first time tried to elucidate the anti-inflammatory potential of aqueous extract of Tectaria cicutaria rhizome (TCR_aq) in vitro as well as in vivo.

Materials and methods

Anti-inflammatory potential of TCR_aq was analyzed in vivo in carrageenan induced rat paw edema model. Serum antioxidant status in TCR_aq-treated as well as untreated control rodents was measured by oxygen radical absorbance capacity (ORAC) assay. In vitro experiments for analyzing the anti-inflammatory potential of TCR_aq were performed on murine macrophage cell line, RAW 264.7. Analysis of nitric oxide release in RAW 264.7 cells was done by Griess reaction. RT-PCR and western blotting experiment was performed to analyze the expression of iNOS. Expression of COX-2 and NFκB proteins was evaluated by western blotting.

Results

TCR_aq significantly reduced the paw volume in Sprague-Dawley rats at a dose of 200 mg/kg body weight, which was comparable with the standard diclofenac treatment. The rats treated with TCR_aq showed a significant increase in the serum antioxidant levels compared to the untreated control animals. TCR_aq was able to reduce the nitric oxide (NO) levels in RAW 264.7 cells that had been stimulated with lipopolysaccharide (LPS). This was accompanied by a corresponding decrease in iNOS expression at mRNA and protein level. Interestingly, TCR_aq was found to decrease the expression of COX-2 as well as the nuclear translocation of NFκB in RAW 264.7 cells.

Conclusion

Our study signifies the anti-inflammatory potential of Tectaria cicutaria and scientifically validates its traditional use in inflammatory conditions.     

柴胡注射液体外抑制PGE2释放的生物活性测定法

目的: 建立rrIL-1β体外干扰下丘脑神经细胞释放PGE₂离体模型,考察柴胡注射作用下该模型中PGE₂含量与药物作用浓度的相关性。 方法: 体外培养下丘脑神经细胞,加入rrIL-1β(40 μg·L^-1)刺激并于不同时间点收集细胞液,ELISA法检测各时间点收集的细胞液中PGE₂含量;体外培养下丘脑神经细胞,加入rrIL-1β刺激后,各组加入不同浓度柴胡注射液,ELISA法检测细胞上清液中PGE₂含量变化,分析加样浓度与对PGE₂抑制率的线性关系。 结果: rrIL-1β能刺激体外培养的下丘脑神经细胞释放PGE₂,并在10 h时达到峰值;柴胡注射液能够显著干扰离体模型中PGE₂的释放(P<0.01,P<0.05),且干扰作用与柴胡注射液作用浓度呈一定的线性相关(r=0.911,P<0.01)。 结论: rrIL-1β能够干扰体外培养的下丘脑神经细胞释放PGE₂,且对PGE₂的抑制生成作用与药物浓度相关性较好。     

An in vitro study on antiproliferative and antiestrogenic effects of Boerhaavia diffusa L. extracts

Ethnopharmacological relevance

Boerhaavia diffusa L. (Nyctinaceae) is a plant of tropical region used in Indian traditional medicine for the treatment of human ailments including abdominal tumor, jaundice, dyspepsia, menstrual disorders, etc. This plant also has antilymphoproliferative, antimetastatic and immunomodulatory effects.

Aim of the study

This study aimed to assess the antiproliferative and antiestrogenic properties of methanol extract of Boerhaavia diffusa (BME) in MCF-7 breast cancer cell lines.

Materials and methods

The effective concentration range of BME on cell viability was analyzed using MTT assay. Hydroxylapatite assay (HAP) was carried out to confirm the competitive binding of BME to the estrogen receptor (ER). The effect of BME on the expression of a selected estrogen responsive gene pS2 was analyzed by RT-PCR. The ability of BME to alter the cell cycle phases and distributions were studied using FACS analysis.

Results

Treatment with varying concentrations of BME (20–320 μg/mL) resulted in moderate to very strong growth inhibition in MCF-7 cell lines. BME competed with [³H]-estradiol for binding to ER with IC₅₀ value of 320 ± 25 μg/mL. RT-PCR analysis revealed that BME reduced the mRNA expression of pS2 indicating the antiestrogenic action of BME. BME treatment for 48 h resulted in a remarkable increase in the number of MCF-7 cells in the G0-G1 fraction from 69.1% to 75.8%, with a reciprocal decrease of cells in all other phases indicating cell cycle arrest at G0-G1 phase.

Conclusions

The results demonstrate that Boerhaavia diffusa possess antiproliferative and antiestrogenic properties and suggest that it may have therapeutic potential in estrogen dependent breast cancers.     

In vitro and in vivo anti-inflammatory effect of Rhodomyrtus tomentosa methanol extract

Ethnopharmacological relevance

Rhodomyrtus tomentosa (Aiton) Hassk. is a representative Thai medicinal plant traditionally used in South Asian countries to relieve various inflammatory symptoms. However, no systematic studies on its anti-inflammatory activity and mechanisms have been reported.

Materials and methods

The effect of the methanol extract from the leaves of this plant (Rt-ME) on the production of inflammatory mediators [nitric oxide (NO) and prostaglandin E₂ (PGE₂)] and the molecular mechanism of Rt-ME-mediated inhibition, including target enzymes, were studied with RAW264.7, peritoneal macrophage, and HEK293 cells. Additionally, the in vivo anti-inflammatory activity of this extract was evaluated with mouse gastritis and colitis models.

Results

Rt-ME clearly inhibited the production of NO and PGE₂ in lipopolysaccharide (LPS)-activated RAW264.7 cells and peritoneal macrophages in a dose-dependent manner. According to RT-PCR, immunoblotting and immunoprecipitation analyses and a kinase assay with mRNA, whole cell extract, and nucleus lysates from RAW264.7 cells and mice, it was revealed that Rt-ME was capable of suppressing the activation of both nuclear factor (NF)-κB and activator protein (AP)-1 pathways by directly targeting Syk/Src and IRAK1/IRAK4.

Conclusion

Rt-ME could have anti-inflammatory properties by suppressing Syk/Src/NF-kB and IRAK1/IRAK4/AP-1 pathways and will be further developed as a herbal remedy for preventive and/or curative purposes in various inflammatory diseases.     

杨桃根多糖体外抗氧化作用的研究

目的:探讨杨桃根多糖(Yangtaogen polysaccharide,YTGP)的抗氧化活性。方法:邻苯三酚自氧化法和邻二氮菲-Fe2+/H2 O2体系研究YTGP对自由基的作用,用硫代巴比妥酸法测小鼠肝中丙二醛(MDA)含量。在体外化学模拟条件下测定杨桃根多糖的总还原能力。结果:YTGP能有效抑制MDA的产生,能明显地抑制.OH和O2-的生成。结论:YTGP在体外有明显的抗氧化作用。     

Mirabijalone E: A novel rotenoid from Mirabilis himalaica inhibited A549 cell growth in vitro and in vivo

Ethnopharmacological relevance

The roots of Mirabilis himalaica have been used in Tibetan folk medicine for treatment of uterine cancer, nephritis edematous, renal calculus and arthrodynia. In our previous work, the ethanol extract of roots had shown potent cytotoxicity against human cancer cells. However, no information is available on the antitumor effect of Mirabilis himalaica. The aim of the present study was to investigate the active constituents guided by bioassay and evaluate the related antitumor efficacy in vitro and in vivo.

Materials and methods

The active subextract (ethyl acetate) was subjected to successive chemical separation using a combination of silica gel, LH-20 chromatography and semi-preparative HPLC. The structures were determined by spectroscopic analysis techniques such as nuclear magnetic resonance (NMR) and mass spectrometry. Three human cancer cell lines, A549, HepG2 and HeLa were used for in vitro cytotoxicity evaluation of all isolated compounds by MTT-assay. Then, the potent and novel compound mirabijalone E was employed to the mechanism study againstA549 cells. BrdU immunofluorescence, soft agar assay and cell cycle analysis were employed to detect the cell proliferation effects. Annexin V-FITC/PI staining assay was used for examining apoptotic effects. Expression levels of apoptosis-related proteins were determined by western blot assay. in vivo tumorigenic assay was used to evaluate the xenograft tumor growth treated with mirabijalone E.

Results

One new rotenoid compound, mirabijalone E, together with eight known rotenoids was isolated from Mirabilis himalaica. Mirabijalone E, 9-O-methyl-inone B, boeravinone C and boeravinone H exhibited cytotoxicity against A 549 and HeLa cells. Further study on mirabijalone E was carried out in vitro and in vivo. Mirabijalone E inhibited A549 cells growth in a time and dose-dependent manner, which arrested cell cycle in S phase. Mechanistically, mirabijalone E treatment resulted in the increase of Bax expression level, the decrease of Bcl-2 level and the activation of caspase-3, which suggested the activation of apoptosis cascades. Consequently, the xenograft treated with mirabijalone E showed markedly suppressed tumor growth.

Conclusions

The result suggested that mirabijalone E, together with active compounds, 9-O-methyl-4-hydroxyboeravinone B, boeravinone C and boeravinone H could be a promising candidate for cancer therapy.     

糙苏素的抗肿瘤活性研究

目的:探讨藏药螃蟹甲中环烯醚萜类成分糙苏素(phlomiol)的抗肿瘤作用.方法:采用MTT法检测糙苏素对体外培养的2种人肿瘤细胞增殖的抑制作用;体内试验采用小鼠实体瘤S180、肝癌H22细胞株接种小鼠,连续给药14 d,计算抑瘤率.采用MTT比色法检测糙苏素对S180荷瘤小鼠NK细胞杀伤活性的影响和对S180荷瘤小鼠T淋巴细胞增殖反应的影响.结果:糙苏素在50～150 mg·L-1对2种肿瘤细胞增殖均有抑制作用,且呈剂量依赖关系(r=0.989,P<0.05);体内试验结果显示小鼠分别灌胃给予2.5,5,10 mg·kg-13个剂量的糙苏素,对接种的实体瘤S180、肝癌H22细胞株的抑瘤率分别为28.5%～65.0%.35.0%～74.5%.同时,糙苏素可显著提高S180荷瘤小鼠淋巴细胞增殖活性(P<0.05),显著增加S180荷瘤小鼠NK细胞的杀伤能力(P<0.05).结论:糙苏素对体外培养的人肿瘤细胞和接种的小鼠肿瘤具有明显抑制作用,能够增强淋巴细胞增殖功能和NK细胞的杀伤能力,具有抗肿瘤和免疫调节功能.