HPLC同时测定穿心莲药材及其注射液中3种内酯类成分的含量

目的利用HPLC同时测定穿心莲药材及其注射液中穿心莲内酯、脱水穿心莲内酯和新穿心莲内酯的含量。方法色谱柱为Shimadzu Shim-pack VP-ODS柱(4.6 mm×150 mm,5μm),以甲醇-水(53∶47)洗脱,流速:1 mL.min?1,柱温:25℃,检测波长225 nm处检测穿心莲内酯,检测波长205 nm处检测脱水穿心莲内酯与新穿心莲内酯。结果穿心莲药材及其注射液中3种内酯类成分能达到很好分离,线性关系良好(r≥0.999 5);平均加样回收率为99.6%~101.2%,RSD<3%。结论本方法灵敏、准确、可靠、重复性好,可用于穿心莲药材及其注射液的质量评价与3种内酯类成分的快速测定。     

Suppression of rat neutrophil reactive oxygen species production and adhesion by the diterpenoid lactone andrographolide

The present study was to examine whether andrographolide, a diterpenoid lactone isolated from the anti-inflammatory herbal medicine Andrographis paniculata (Burm. f.) Nees. (Acanthaceae), has the ability to prevent phorbol-12-myristate-13-acetate (PMA)-induced reactive oxygen species (ROS) production, as well as N-formyl-methionyl-leucyl-phenylalanine (fMLP)-induced adhesion by rat neutrophils. Results demonstrated that PMA (100 ng/ml) induced rapid accumulation of H2O2 and O2. in neutrophils within 30 minutes. Andrographolide (0.1 to 10 microM) pretreatment (10 min, 37 degrees C) significantly attenuated the accumulation of these two oxygen radical metabolites. Administration of andrographolide also significantly prevented fMLP-induced neutrophil adhesion. These data suggest that preventing ROS production and neutrophils adhesion may confer andrographolide the ability to be an anti-inflammatory drug.     

Andrographolide mitigates cardiac apoptosis to provide cardio‐protection in high‐fat‐diet‐induced obese mice

Excessive intake of high fat diet (HFD) and associated obese conditions are critical contributors of cardiac diseases. In this study, an active metabolite andrographolide from Andrographis paniculata was found to ameliorate HFD‐induced cardiac apoptosis. C57/BL6 mouse were grouped as control (n = 9), obese (n = 8), low dose (25 mg/kg/d) andrographolide treatment (n = 9), and high dose (50 mg/kg/d) andrographolide treatment (n = 9). The control group was provided with standard laboratory chow and the other groups were fed with HFD. Andrographolide was administered through oral gavage for 1 week. Histopathological analysis showed increase in apoptotic nuclei and considerable cardiac‐damages in the obese group signifying cardiac remodeling effects. Further, Western blot results showed increase in pro‐apoptotic proteins and decrease in the proteins of IGF‐1R‐survival signaling. However, feeding of andrographolide significantly reduced the cardiac effects of HFD. The results strongly suggest that andrographolide supplementation can be used for prevention and treatment of cardiovascular disease in obese patients.     

蓝心草颗粒的定性定量方法研究

目的 建立蓝心草颗粒的定性定量方法.方法 对蓝心草颗粒中板蓝根、穿心莲、鱼腥草进行薄层色谱鉴别.用HPLC法测定穿心莲内酯、脱水穿心莲内酯的含量,采用Dikma DiamonsiL C18柱,以甲醇-水(48∶52)为流动相,流速为1.0 mL·min -1,检测波长为240 nm.结果 薄层色谱斑点清晰,阴性样品无干扰.穿心莲内酯进样量在0.036 6～1.464μg与峰面积线性关系良好,r=0.999 2,平均回收率为97.6％,RSD为1.3％(n=9);脱水穿心莲内酯进样量在0.041 9～1.677 μg与峰面积线性关系良好,r=0.999 9,平均回收率为98.9％,RSD为1.7％(n=9).结论 本法专属性强,准确度高,重现性好,可作为该产品质量控制方法.     

Effect of Andrographolide and Ethanol Extract of Andrographis paniculata on Liver Glycolytic,Gluconeogenic, and Lipogenic Enzymes in a Type 2 Diabetic Rat Model

Andrographis paniculata (Burm.f.) Nees (Acanthaceae) (AP), containing the major active principle, andrographolide,(AG) has been used ethnomedically in the treatment of diabetes and hypertension in Malaysia and India. Type 2 diabetes mellitus was induced by administering nicotinamide (NA) 180 mg/kg i.p., followed by streptozotocin (STZ) 45 mg/kg i.p., 15 min later and allowed for 15 days to develop. Diabetic rats treated with active principle andrographolide 10 mg/kg and ethanol extract of Andrographis paniculata at doses of 500 and 1000 mg/kg, p.o, for 21 days, caused a significant reduction in fasting serum glucose levels on all days analysed (P < 0.01). Treatment with andrographolide and extract showed an increase in glucokinase (GK; P < 0.05), hexokinase (HK; P < 0.05), and lactate dehydrogenase (LDH; P < 0.05) enzyme levels respectively. Significant reductions were observed in serum cholesterol (P < 0.05), triglycerides (P < 0.05), free fatty acids (P < 0.05), and liver glucose 6-phosphatase (G6P’Tase; P < 0.05) enzyme levels on treatment with andrographolide and extract. Glucose 6-phosphate dehydrogenase (G6PDH; P < 0.05) level also showed a significant increase on treatment with andrographolide and extract. Liver antioxidant status (P < 0.05) also improved significantly on treatment with andrographolide and extract. However, no significant increase in serum insulin was found on treatment with either andrographolide or extract, suggesting an effective extra pancreatic mechanism. Thus, the present study demonstrates that andrographolide at 10 mg/kg, and ethanol extract of Andrographis paniculata at 500 and 1000 mg/kg, p.o., on treatment for 21 days, exhibited antidiabetic, hypolipidemic, and antioxidant activities in an adult streptozotocin-nicotinamide type 2 diabetes mellitus model. The results support the traditional use of Andrographis paniculata in the treatment of type 2 diabetes mellitus, and its associated plethora of complications.     

A Rapid Method for Isolation of Andrographolide from Andrographis Paniculata Nees (Kalmegh)

A simple and rapid method for isolation of andrographolide from the leaves of Andrographis paniculata is reported. This involves extraction of the leaf powder by cold maceration in a 1:1 mixture of dichloromethane and methanol and isolation of andrographolide directly from the resulting extract by recrystallisation. The identity of the compound was confirmed through IR, UV, mass and melting point, and co-chromatography with a reference standard on TLC. The purity of the compound was confirmed by TLC, UV absorption spectrum, HPLC and differential scanning calorimetry, the latter of which gave the melting point of andrographolide as 235.3°C.     

A rapid method for isolation of andrographolide from andrographis paniculata nees (kalmegh)

A simple and rapid method for isolation of andrographolide from the leaves of Andrographis paniculata is reported. This involves extraction of the leaf powder by cold maceration in a 1:1 mixture of dichloromethane and methanol and isolation of andrographolide directly from the resulting extract by recrystallisation. The identity of the compound was confirmed through IR, UV, mass and melting point, and co-chromatography with a reference standard on TLC. The purity of the compound was confirmed by TLC, UV absorption spectrum, HPLC and differential scanning calorimetry, the latter of which gave the melting point of andrographolide as 235.3°C.     

妇科千金分散片质量标准研究

目的妇科千金分散片的质量标准研究。方法采用薄层色谱法鉴别当归、穿心莲、党参,用高效液相色谱法测定穿心莲内酯和脱水穿心莲内酯的含量。SHIMADZU VP－ODS（150×4．6mm,5μm）柱,以甲醇：水=52：48为流动相．穿心莲内酯检测波长为225nm,脱水穿心莲内酯检测波长为254nm。结果能准确对当归、穿心莲、党参进行定性鉴别;穿心莲内酯与脱水穿心莲内酯分别在0．2004－2．0400μg与0．1172－0．7024μg呈良好线性关系。结论本法简便、快速、准确,灵敏,可作为该制剂质量控制的有效方法。     

不同提取方法对穿心莲内酯含量的影响

目的:探讨穿心莲的最佳提取方法。方法:以穿心莲内酯为考察指标,以HPLC法测定不同提取方法对穿心莲内酯含量及出膏率的影响。结果:穿心莲内酯含量以醇提法及．0．01％碱水提取为佳,含量分别为4．43％和4．31％,但出膏率后者较高,达24．08％。结论:穿心莲的提取方法以0．01％碱水回流提取为最佳。     

Gender-associated modulation of inducible CYP1A1 expression by andrographolide in mouse liver

We previously observed a strong synergistic effect on polycyclic aromatic hydrocarbon (PAH)-induced CYP1A1 expression by andrographolide, a major constituent of an herbal medicine derived from the plant Andrographis paniculata, in mouse hepatocytes in primary culture. The present paper describes confirmation of an enhancing effect of andrographolide on the CYP1 family in vivo in the PAH-responsive C57BL/6 mouse. Andrographolide did not alter CYP1 expression in the PAH-nonresponsive DBA/2 mouse. The enhanced expression induced by andrographolide was observed in male C57BL/6 mice, but not in intact or ovariectomized females, or in orchiectomized male mice. However, treatment with testosterone restored the effect in both orchiectomized males and ovariectomized females. These observations indicate a male hormone-related system to be a crucial mediator of the modulation of CYP1 expression by andrographolide. Precautions should be taken regarding the use of A. paniculata as an alternative medication or health promotion, according to its distinctive characterization on sexually dimorphic modulation of CYP1A1 expression.     

Choleretic effect of andrographolide in rats and guinea pigs.

Andrographolide from the herb Andrographis paniculata (whole plant) per se produces a significant dose (1.5-12 mg/kg) dependent choleretic effect (4.8-73%) as evidenced by increase in bile flow, bile salt, and bile acids in conscious rats and anaesthetized guinea pigs. The paracetamol induced decrease in volume and contents of bile was prevented significantly by andrographolide pretreatment. It was found to be more potent than silymarin, a clinically used hepatoprotective agent.     

穿心莲浸膏片的质量研究

本文对甲、乙、丙、丁四个生产厂的穿心莲浸膏片的外观、水分、硬度、崩解度、穿心莲总内脂含量、溶出度进行了测定,并对穿心莲总内酯的含量分布进行了正态检验。结果表明:甲、丙两厂产品质量较好,乙厂次之,丁厂较差。     

Antihyperglycemic effect of andrographolide in streptozotocin-induced diabetic rats

The antihyperglycemic action of andrographolide, an active principle in the leaves of Andrographis paniculata (Burm. f.) Nees, was investigated in streptozotocin-induced diabetic rats (STZ-diabetic rats). Oral treatment of andrographolide decreased the plasma glucose concentrations of STZ-diabetic rats in a dose-dependent manner. Similar treatment with andrographolide also decreased the plasma glucose in normal rats and the maximal effect was more marked than that in STZ-diabetic rats. Andrographolide at the effective dose (1.5 mg/kg) significantly attenuated the increase of plasma glucose induced by an intravenous glucose challenge test in normal rats. In the isolated soleus muscle of STZ-diabetic rats, andrographolide enhanced the uptake of radioactive glucose in a concentration-dependent manner. Moreover, the mRNA and protein levels of the subtype 4 form of the glucose transporter (GLUT4) in soleus muscle were increased after repeated intravenous administration of andrographolide in STZ-diabetic rats for 3 days. These results suggest that andrographolide can increase the glucose utilization to lower plasma glucose in diabetic rats lacking insulin.     

基于多组分分析的痢泻灵片质量标准研究

目的 建立痢泻灵片的质量标准。方法 采用薄层色谱法对痢泻灵片中拳参、穿心莲、苦参进行定性鉴别；采用HPLC对痢泻灵片中穿心莲类成分和苦参碱类成分进行测定。结果 拳参、穿心莲、苦参薄层斑点清晰，分离效果良好，重复性好。穿心莲内酯、新穿心莲内酯、14-去氧穿心莲内酯、脱水穿心莲内酯、苦参碱、氧化苦参碱、槐果碱、氧化槐果碱在各自的浓度范围内线性关系良好，仪器精密度、重复性、稳定性、加样回收率的RSD值均符合要求。结论 建立的方法简单易操作，可用于控制痢泻灵片的质量。     

Antidiabetic and antihiperlipidemic effect of Andrographis paniculata (Burm. f.) Nees and andrographolide in high-fructose-fat-fed rats

Objectives:

Andrographis paniculata (Burm. f.) Nees originates from India and grows widely in many areas in Southeast Asian countries. Andrographis paniculata (Burm. f.) Nees has shown an antidiabetic effect in type 1 DM rats. The present study investigates the purified extract of the plant and its active compound andrographolide for antidiabetic and antihyperlipidemic effects in high-fructose-fat-fed rats, a model of type 2 DM rats.

Materials and Methods:

Hyperglycemia in rats was induced by high-fructose-fat diet containing 36% fructose, 15% lard, and 5% egg yolks in 0.36 g/200 gb.wt. 55 days. The rats were treated with the extract or test compound on the 50^th day. Antidiabetic activity was measured by estimating mainly the pre– and postprandial blood glucose levels and other parameters such as cholesterol, LDL, triglyceride, and body weight.

Results:

The purified extract and andrographolide significantly (P<0.05) decreased the levels of blood glucose, triglyceride, and LDL compared to controls. However, no changes were observed in serum cholesterol and rat body weight. Metformin also showed similar effects on these parameters.

Conclusions:

Andrographis paniculata (Burm. f.) Nees or its active compound andrographolide showed hypoglycemic and hypolipidemic effects in high-fat-fructose-fed rat.KEY WORDS: Andrographis paniculata (Burm. f.) Nees, andrographolide, diabetes, fructose, hyperlipidemia     

穿黄消炎胶囊的制备及质量标准研究

目的建立穿黄消炎胶囊的定性定量分析方法。方法采用薄层色谱法定性鉴别制剂中的穿心莲和一枝黄花,采用高效液相色谱法测定制剂中脱水穿心莲内酯的含量。结果薄层色谱法可定性检出穿心莲内酯与脱水穿心莲内酯;脱水穿心莲内酯质量浓度在0.03～0.15g/L范围内与峰面积线性关系良好,r=0.9999,平均回收率为98.44%,RSD为0.72%（n=5）。结论所用定性定量方法可用于穿黄消炎胶囊的质量控制。     

Suppression of NO production in activated macrophages in vitro and ex vivo by neoandrographolide isolated from Andrographis paniculata

In this study, we investigated the in vitro and ex vivo suppressive effects of Andrographis paniculata on nitric oxide (NO) production in mouse peritoneal macrophages elicited by bacillus Calmette-Guéin (BCG) and stimulated by lipopolysaccharide (LPS). Incubation of BCG-induced macrophages with the methanol extract of A. paniculata reduced LPS stimulated NO production. The diterpene lactones andrographolide and neoandrographolide were isolated as active components from the extract. These compounds suppressed NO production in a concentration-dependent manner in the concentration range from 0.1 to 100 microM and their IC50 values were 7.9 and 35.5 microM. Neoandrographolide also suppressed NO production by 35 and 40% when the macrophages were collected after oral administration of neoandrographolide at doses of 5 and 25 mg/kg/d and LPS stimulated NO production was examined. However, andrographolide did not reduce NO production on oral administration at the same doses. These results indicate that neoandrographolide, which inhibited NO production both in vitro and ex vivo may play an important role in the use of A. paniculata as an anti-inflammatory crude drug.