Presence Of Aristolochic Acid Research Articles

Toxicological Safety Evaluation in Acute and 21-Day Studies of Ethanol Extract from Solanum lyratum Thunb.

Solanum lyratum (Solanaceae) is a traditional Chinese medicine widely used to remedy cold fever, damp-heat jaundice, herpes, and nephritis dropsy. Despite its obvious therapeutic advantages, few toxicological studies have involved the efficacy and safety of its long-term treatment. To investigate the acute and subchronic toxicity of the extract of 75% ethanol extract of whole Solanum lyratum (ESL) after oral administration in mice. In acute toxicity experiment, mice were intragastric administration with ESL at doses of 1000, 2000, 3000, 4000, or 5000 mg/kg for 1 day. In a subchronic toxicity experiment, mice were intragastrically administration with ESL at doses of 180, 360, and 720 mg/kg and 0.9% saline for 21 days. Weight gain, hematological, biochemical, and histopathological analysis of vital organs were evaluated. The presence of aristolochic acid I in ESL was studied using UPLC-QTOF-MS. Phytochemical analysis indicated that the presence of aristolochic acid I in ESL was 0.0025 mg/g. This relatively low concentration is not enough to cause toxicity. In the acute toxicity experiment, neither mortality nor clinical alterations were shown, except for the mild transient diarrhea at 5000 mg/kg. So the LD50 value of ESL was assessed to be more than 5000 mg/kg. In the subchronic toxicity experiment, neither mortality nor treatment-related clinical signs were observed. There was a significant increase in body weight, hemoglobin (HB), and urea nitrogen (BUN) after administration with ESL at 180 mg/kg. In addition, the weight of the stomach was increased and the hematocrit (HCT) was decreased after administration with ESL at 360 mg/kg. The changes were not considered treatment-related toxicological effects because the toxicity and histopathological analysis indicate that the extracts are safe for oral administration.

Chemical Constituents and Pharmacology properties of Aristolochia triangularis: a south brazilian highly-consumed botanical with multiple bioactivities.

Aristolochia triangularis Cham., is one of the most frequently used medicinal plant in Southern Brazil. Preparations containing the leaves and/or stems are traditionally used as anti-inflammatory, diuretic, as well as antidote against snakebites. This study screened A. triangularis extracts, fractions and isolated compounds for different bioactivities. A weak antiproliferative activity against human lung cancer cell line (A549) was observed only for chloroform fraction obtained from stems (CFstems - CC50: 2.93 µg/mL). Also, a moderate antimicrobial activity against Staphylococcus aureus was detected just for chloroform fraction obtained from leaves (CFleaves -13-16 mm inhibition zone). Additionally, two semi-purified fractions (CFstems-4 and CFleaves-4) selectively inhibited HSV-1 replication (IC50 values of 0.40 and 2.61 µg/mL, respectively), while only CFleaves showed promising results against Leishmania amazonensis. Fractionation of extracts resulted in the isolation of one neolignan (-) cubebin and one lignan (+) galbacin. However, these compounds are not responsible for the in vitro bioactivities herein detected. The presence of aristolochic acid I and aristolochic acid II in the crude ethanol extract of stems (CEEstems) and leaves (CEEleaves) was also investigated. The HPLC analysis of these extracts did not display any peak with retention time or UV spectra comparable to aristolochic acids I and II.

Sequestration of Aristolochic Acid I from Aristolochia pilosa by Mapeta xanthomelas Walker, 1863

Sequestration of secondary plant chemicals and brightly colored bodies occur in a number of unpalatable insects. The utilization of toxic plant chemicals has been proposed as a strategy of chemical defense, while aposematic coloration may advertise unpalatability. Here, we tested for the presence of aristolochic acid I in leaves of Aristolochia pilosa and female bodies of Mapeta xanthomelas, obtained from larvae feeding on the plant, using high performance liquid chromatography with photodiode array detection and electrospray ionization mass spectrometry. The presence of aristolochic acid I in females of this conspicuous diurnal moth, an oligophagous herbivore of Aristolochia, is the first report of sequestration of aristolochic acids by an herbivore other than a species of Papilionidae.

A Sensitive Enzyme-Linked Immunosorbent Assay for Detecting Carcinogenic Aristolochic Acid in Herbal Remedies

Aristolochic acid, a naturally occurring nephrotoxin and rodent carcinogen, has been associated with the development of various nephropathies in humans. Developing a sensitive and rapid method to screen the aristolochic acid levels in herbal remedies is urgent for protecting public health. Polyclonal antibodies for aristolochic acid were generated from rabbits after the animals had been immunized with either aristolochic acid-ovalbumin (OVA) or aristolochic acid-keyhole limpet hemocyanin (KLH). A competitive indirect enzyme-linked immunosorbent assay (ciELISA) and a competitive direct enzyme-linked immunosorbent assay (cdELISA) were used for the characterization of the antibodies and for analysis of aristolochic acid contaminated in herbal medicine and diet pills. The antibody titers in the serum of rabbits immunized with aristolochic acid-OVA were considerably higher than those from aristolochic acid-KLH-immunized rabbits. The antibodies from the aristolochic acid-OVA-immunized rabbits were further characterized. In the ciELISA with aristolochic acid-KLH as the plate-coating antigen, the concentrations of the aristolochic acid mixture, aristolochic acid I, and aristolochic acid II that caused 50% inhibition (IC50) of binding of antibodies to aristolochic acid-KLH were found to be 1.2, 0.7, and 18 ng/mL, respectively. When 0.25-5 microg/g of standard aristolochic acid was spiked to ground lotus seeds and then extracted with 0.01 M phosphate-buffered saline, the recovery rate was found to be 86.5% in the ciELISA. Analysis of aristolochic acid in herbal medicine and diet pills with ciELISA showed that 10 of the 12 examined samples were contaminated at levels from 0.6 to 655 microg/g. The presence of aristolochic acid was also confirmed by the high-performance liquid chromatography method.

Detection of aristolochic acid in Chinese phytomedicines and dietary supplements used as slimming regimens

Over the last 10 years, numerous cases of intoxications, leading for the most part to end-stage renal failure, have been reported after consumption of slimming regimens made of Chinese herbal preparations. These intoxications were associated with species of the Aristolochia genus, such as Aristolochia fangchi (Aristolochiaceae), known to contain very nephrotoxic and carcinogenic metabolites named aristolochic acids. Several commercial dietary supplements, teas and phytomedicines used as slimming regimens were analysed for their aristolochic acid I content. A preliminary detection of this toxic compound was made by thin-layer chromatography. The presence of aristolochic acid I in these preparations was confirmed by a HPLC/UV-DAD/MS analysis. A quantitative determination of aristolochic acid I was also achieved in the incriminated preparations using both UV and MS detection. Out of 42 analysed preparations, four were found to contain aristolochic acid I and two were suspected to contain aristolochic acid derivatives. Immediate removal of these products from the Swiss market was called for.

Chinese Herb Nephropathy

In 1994, a 44-year-old woman progressed from normal renal function to advanced renal failure and end-stage renal disease within 8 months. Biopsy revealed extensive interstitial fibrosis with focal lymphocytic infiltration. She received a cadaveric renal transplant in January 1996 and had an uneventful posttransplant course. As a result of a minor motor vehicle accident, the patient had received acupuncture and Chinese herbal medicine for pain relief approximately 5 months before the onset of renal symptoms. After the transplant, analysis of the herbal remedies clearly indicated the presence of aristolochic acid in 2 of the 6 Chinese herbs ingested. Ingestion of aristolochic acid has been linked to a newly defined entity, Chinese herb nephropathy (CHN). This article discusses the history of CHN and its implication in the current case and in other recent similar cases and makes recommendations to avoid future problems caused by unregulated use of herbal medicines. This is the first reported case of CHN in the USA.

Chinese Herb Nephropathy

In 1994, a 44-year-old woman progressed from normal renal function to advanced renal failure and end-stage renal disease within 8 months. Biopsy revealed extensive interstitial fibrosis with focal lymphocytic infiltration. She received a cadaveric renal transplant in January 1996 and had an uneventful posttransplant course. As a result of a minor motor vehicle accident, the patient had received acupuncture and Chinese herbal medicine for pain relief approximately 5 months before the onset of renal symptoms. After the transplant, analysis of the herbal remedies clearly indicated the presence of aristolochic acid in 2 of the 6 Chinese herbs ingested. Ingestion of aristolochic acid has been linked to a newly defined entity, Chinese herb nephropathy (CHN). This article discusses the history of CHN and its implication in the current case and in other recent similar cases and makes recommendations to avoid future problems caused by unregulated use of herbal medicines. This is the first reported case of CHN in the USA.

25 The cytotoxicity of aristolochic acid in the MDCK and LLC-PK1 renal cell lines

Rapidly progressive fibrosing interstitial nephritis in young women following a slimming regimen including Chinese herbs was ascribed to the administration of Aristolochiafangchi instead of Stephania tetrandra. This was concluded from the presence of aristolochic acid (AA) in the capsules used (Vanhaelen et al., Lancet 343, 174, 1994). Here we report the in vitro cytotoxicity of AA in the MDCK (canine distal) and LLC-PK1 (porcine proximal) renal cell lines of tubular origin. A commercially available mixture of 52% AAI and 41% AAII was used to measure the neutral red uptake inhibition. The results were quantifed by the NI50, i.e. the concentration (in mg/l) of AA inducing a 50% inhibition in neutral red uptake. The NI50 values are given in the table. The cyto-

Biosynthesis of aristolochic acid.

Some Aristolochia species were investigated for the presence of aristolochic acid and magnoflorine. In all cases these two compounds were found in the roots and rhizomes of the plants