Interaction of five anthraquinones from rhubarb with human organic anion transporter 1 (SLC22A6) and 3 (SLC22A8) and drug–drug interaction in rats

Ethnopharmacological relevance

Rhubarb is a well-known traditional Chinese medicine and has been used in China for thousands of years. Anthraquinone derivatives including rhein, emodin, aloe-emodin, chrysophanol and physcion are the important components in rhubarb.

Materials and methods

Here we studied the interaction of five anthraquinone derivatives with human renal organic anion transporter 1 (hOAT1) and hOAT3 stably expressed in cells, and interaction of rhein or rhubarb extract (RE) with furosemide (FS, substrate of OATs) in rats.

Results

Uptake of 6-carboxyl fluorescein via hOAT1 and fluorescein via hOAT3 were markedly inhibited by rhein, emodin and aloe-emodin, and slightly inhibited by chrysophanol and physcion. The estimated IC₅₀ values for rhein, emodin, aloe-emodin and probenecid (typical inhibitor of hOAT1 and hOAT3) were 0.23, 0.61, 2.29 and 18.34 μM for hOAT1, and 0.08, 1.22, 5.37 and 5.83 μM for hOAT3, respectively. Furthermore, the data from the cellular accumulation assay indicated that these five compounds were not substrates of hOAT1 or hOAT3. Pharmacokinetic interaction between rhein and FS in rats showed that area under the curve (AUC_0–t) for FS was increased by 65% when coadministrated with rhein. RE was also used to interact with FS in rats and results showed that AUC_0–t of FS was increased by 32% and by 52% when coadministrated with single-dose or multiple-dose of RE, respectively.

Conclusions

These findings suggested that five anthraquinones inhibited hOAT1 and hOAT3, but these compounds were not transported by hOAT1 or hOAT3. Furthermore, rhein or RE, might cause drug–drug interaction when coadministrated with substrates of OAT1 or OAT3 in vivo.     

Pharmacokinetic study of salvianolic acid A in beagle dog after oral administration by a liquid chromatography–mass spectrometry method: A study on bioavailability and dose proportionality

Ethnopharmacological relevance

Salvianolic acid A (SAA) is one of the main water-soluble components isolated from Salvia miltiorrhiza Bunge. Pharmacological researches revealed that it had various curative activities after oral and intravenous administration, including beneficial effects on diabetes and its complications, cardioprotective effect, anti-platelet aggregation, and so on. However, there is no report regarding the pharmacokinetics of SAA in beagle dogs after oral administration up to now.

Aim of the study

To study the pharmacokinetics of different doses of SAA in beagle dogs and figure out the absolute bioavailability and dose proportionality of SAA after oral administration.

Materials and methods

Male and female beagle dogs were orally administered SAA 5, 10 and 20 mg/kg randomly. The plasma drug concentration was detected by a rapid, sensitive and reproducible liquid chromatography–mass spectrometry (LC–MS) method. The pharmacokinetic parameters were calculated from plasma concentration–time data using the DAS pharmacokinetic software Data Analysis System Version 3.0 program.

Results

After single-dose oral administration of SAA, the mean peak plasma concentration (C_max) values for groups treated with 5, 10 and 20 mg/kg doses ranged from 14.38 to 38.18 µg/L, and the mean area under the concentration–time curve (AUC_(0–t)) values ranged from 38.77 to 130.33 (µg/L⋅ h). SAA showed lack of dose proportionality over the dose range 5–20 mg/kg, based on the power model. However, the increase in systemic exposure with dose appeared linear. The absolute bioavailability was calculated to range from 1.47% to 1.84%.

Conclusion

The pharmacokinetic properties of SAA in beagle dogs after oral administration were characterized as rapid oral absorption, quick clearance, and poor absolute bioavailability. Systemic exposure exhibited lack of dose proportionality over the dose range 5–20 mg/kg. Furthermore, a readily preparative LC–MS method was demonstrated in this study for the research of traditional Chinese medicine.     

Greater habitual resveratrol intakes were associated with lower risk of hip fracture- a 1:1 matched case–control study in Chinese elderly

The aim of the study was to testify the association of dietary resveratrol (RSV) intakes with hip fracture risk in Chinese elderly. This was a 1:1 age- and gender- matched case–control study. Eligible cases were newly diagnosed patients of hip fracture. Dietary assessment was made by a 79-item validated food frequency questionnaire. Habitual RSV intakes were estimated as the sum of trans- and cis- isomers of resveratrol and piceid according to the available database. Multivariable conditional logistic regression was applied to examine the relationship of dietary RSV and RSV-rich foods with hip fracture risk. A total of 1,070 pairs of hip fracture incident cases and controls were recruited and 1,065 were included for analysis. Compared with the lowest group, total RSV in the highest quartile group had significantly reduced hip fracture risk by 66.3% (OR: 0.337, 0.222 ~ 0.571, p_trend < 0.001). Similar findings were observed for cis- and trans-RSV, cis- and trans-Piceid, as well as RSV-rich foods (grapes, apples and nuts) respectively. Subgroup analysis suggested more evident findings among female and less obese participants. Our findings demonstrated that higher habitual RSV intakes and RSV-rich foods, even in a relatively low amount, were associated with reduced risk of hip fracture in Chinese elderly.     

A modified preparation (LMK03) of the oriental medicine Jangwonhwan reduces Aβ1–42 level in the brain of Tg-APPswe/PS1dE9 mouse model of Alzheimer disease

Ethnopharmacological relevance

The oriental medicine Jangwonhwan, which is a boiled extract of 12 medicinal herbs/mushroom, has been prescribed for patients with cognitive dysfunction. Recently, a modified recipe of Jangwonhwan (LMK02-Jangwonhwan) consisting of seven medicinal plants/mushroom, was shown to have a therapeutic potential to ameliorate AD-like pathology.

Aim of the study

It was investigated whether a further reduction of Jangwonhwan (LMK03-Jangwonhwan) retains the potency to suppress the AD-like pathology.

Materials and Methods

The transgenic mice of Alzheimer disease, Tg-APPswe/PS1dE9, were fed LMK03-Jangwonhwan consisting of two of the herbs, white Poria cocos (Schw.) Wolf and Angelica gigas Nakai, which could protect the AD-like pathology at 300 mg/kg/day of dose for 3 months. In vitro cell biological study, immunohistological and ELISA (enzyme-linked immunosorbent assay) analyses were used to assess its neuroprotective effects against Aβ-induced cell death, and the Aβ accumulation and plaque deposition in the brain.

Results

In vitro study with SH-SY5Y neuroblastoma cells showed that LMK03-Jangwonhwan could protect from cytotoxicity induced by hydrogen peroxide or oligomeric Aβ_1–42. Tg-APPswe/PS1dE9 mice were administered LMK03-Jangwonhwan at 300 mg/kg/day for 3 months from 4.5 months of age. Immunohistological and ELISA analyses showed that LMK03-Jangwonhwan partially reduced Aβ_1–42 and Aβ_1–40 levels and β-amyloid plaque deposition in the brain of Tg-APPswe/PS1dE9 mice. However, LMK03-Jangwonhwan poorly suppressed accumulation of reactive oxidative stress in the hippocampus of Tg-APPswe/PS1dE9 mice and inefficiently improved the expression of phospho-CREB and calbindin, the cellular factors that were down-regulated in AD-like brains.

Conclusions

These results suggest that LMK03-Jangwonhwan has a potency to inhibit AD-like pathology at a detectable level, but LMK03 is not likely to retain the major ability of LMK02-Jangwonhwan to modify AD pathology in several AD-related molecular parameters.     

Zhengyuan capsule(正元胶囊)for the treatment of cancer-related fatigue in lung cancer patients undergoing operation:a study proto-col for a randomized controlled trial

OBJECTIVE:To evaluate the efficacy and safety of Zhengyuan capsule(正元胶囊)when treating Can-cer-related fatigue(CRF)in lung cancer patients un-dergoing surgical o...     

Metabolomics study of biochemical changes in the serum and articular synovium tissue of moxibustion in rats with collagen-induced arthritis: 艾灸对胶原诱导性关节炎大鼠血清和关节滑膜组织生化改变的代谢组学研究

ObjectiveTo demonstrate the rheumatoid arthritis (RA) mechanisms by determining the biochemical changes. To investigate the therapeutic mechanism of moxibustion in RA-model rats using a gas chromatography-mass spectrometry (GC–MS) metabolomics approach.MethodsA total of 24 rats were divided into three groups as follows: normal control group, model group and moxibustion group. Rats in model group and moxibustion group were set up collagen-induced arthritis (CIA) model. Rats in moxibustion group were treated by moxibustion. After 3 weeks of intervention, right ankle joint, serum and articular synovium samples were collected. Right ankle joint samples were used for histopathological evaluation between 3 groups to get the pathological changes of tissues and cells. Serum and articular synovium samples were used to analyze the changed metabolites of moxibustion on RA rats by the GC–MS based metabolomics.ResultsTreatment of moxibustion not only significantly increased the weight of CIA rats, reduced the swelling of hind paw, arthritic scores, IL-1β, TNF-α but also improved histopathological evaluation in right ankle joint samples. Sixteen significantly altered metabolites were found in RA rats as potential biomarkers of arthritis. Thirteen metabolites, significantly adjusted by moxibustion to help relieve arthritis, were selected out as biomarkers of antiarthritic mechanism of moxibustion, which were mainly involved in phenylalanine, tyrosine and tryptophan biosynthesis, glycine, serine and threonine metabolism, phenylalanine metabolism, alanine, aspartate and glutamate metabolism, glyoxylate and dicarboxylate metabolism and aminoacyl-tRNA biosynthesis.ConclusionsWe have indicated moxibustion treatment is able to resist inflammation in CIA rats effectively. Using GC–MS metabolomics technique, we detect novel metabolites in the moxibustion antiarthritic process, which may aid in advanced understanding of arthritis and therapeutic mechanism of moxibustion.     

A search for natural bioactive compounds in Bolivia through a multidisciplinary approach. Part III. Evaluation Of the antimalarial activity of plants used by Alteños Indians

A total of 40 plant extracts traditionally used by the Alte?os Indians, a native community living between the Andean block and the tropical valleys of Bolivia, were screened for antimalarial activity in vitro on Plasmodium falciparum chloroquine resistant (Indo) strain, and in vivo on rodent malaria Plasmodium vinckei petteri. Eleven extracts displayed good or moderate activity in vivo, and ten extracts good or very good antimalarial activity in vitro. Results of the screening are discussed here, in relation with the traditional use of plants.     

Oriental medicine Jangwonhwan reduces Aβ(1–42) level and β-amyloid deposition in the brain of Tg-APPswe/PS1dE9 mouse model of Alzheimer disease

Ethnopharmacological relevance

Jangwonhwan, a boiled extract of 12 medicinal plants/mushroom including Korean red ginseng (Panax ginseng C.A. Meyer), has been prescribed for patients with cognitive dysfunction and are believed to induce brain activity enhancement, provide light sedation, and facilitate sound sleep.

Aim of the study

The present study was carried out to investigate whether Jangwonhwan has a beneficial effect on the brain of Alzheimer disease.

Materials and methods

The transgenic mice of Alzheimer disease, Tg-APPswe/PS1dE9, were fed a modified recipe of Jangwonhwan consisting of a boiled extract of 7 herbs/mushroom (called LMK02-Jangwonhwan) at 400 mg/kg/day of dose for 3 months from 4.5 months of age. Immunohistological and ELISA analyses were used to assess the Aβ accumulation and plaque deposition in the brain. Other in vitro and in vivo works were performed to understand the underlying mechanism.

Results

LMK02-Jangwonhwan notably reduced Aβ(1–42) and Aβ(1–40) levels, concomitantly with a reduction of plaque deposition, in the brain of Tg-APPswe/PS1dE9 mice. LMK02-Jangwonhwan partially suppressed oxidative stress accumulation, and prevented the down-regulation of phospho-CREB and calbindin typically seen in the hippocampus of AD-like brains. In vitro study with SH-SY5Y neuroblastoma cells showed that LMK02-Jangwonhwan inhibited oxidative stress and Aβ-induced neurotoxicity.

Conclusion

The present study suggests that LMK02-Jangwonhwan confers a therapeutic potential to ameliorate AD-like pathology in the brain of Tg-APPswe/PS1dE9 mice.     

Is aristolochic acid nephropathy a widespread problem in developing countries?: A case study of Aristolochia indica L. in Bangladesh using an ethnobotanical–phytochemical approach

Ethnopharmacological relevance

Species of Aristolochia are associated with aristolochic acid nephropathy (AAN), a renal interstitial fibrosis and upper urinary tract cancer (UUC). Aristolochic acid nephropathy has been reported in ten countries but its true incidence is unknown and most likely underestimated. By combining an ethnobotanical and phytochemical approach we provide evidence for the risk of AAN occurring in Bangladesh. More specifically, we assess the intra-specific variation of aristolochic acid analogues in medicinally used Aristolochia indica samples from Bangladesh.

Materials and methods

Ethnobotanical information was collected from 16 kavirajes (traditional healers) in different study locations in Bangladesh. Plant samples were obtained from native habitats, botanical gardens, herbal markets and pharmaceutical companies. The samples were extracted using 70% methanol and were analysed using LC-DAD-MS and ¹H-NMR.

Results

Roots as well as leaves are commonly used for symptoms such as snake bites and sexual problems. Among the informants knowledge about toxicity or side effects is very limited and Aristolochia indica is often administered in very high doses. Replacement of Aristolochia indica with other medicinal plants such as Rauvolfia serpentina (L.) Benth. ex Kurz was common. Aristolochia indica samples contained a variety of aristolochic acid analogues such as aristolochic acid I, aristolochic acid II, cepharadione A and related compounds.

Conclusions

AAN cases are likely to occur in Bangladesh and more awareness needs to be raised about the health risks associated with the use of Aristolochia indica and other species of Aristolochia as herbal medicines.     

Dentatin isolated from Clausena excavata induces apoptosis in MCF-7 cells through the intrinsic pathway with involvement of NF-κB signalling and G0/G1 cell cycle arrest: A bioassay-guided approach

Ethnopharmacological relevance

Clausena excavata Burm. f. has been used in folk medicines in eastern Thailand for the treatment of cancer.

Materials and methods

To investigate the apoptosis mechanism, we isolated dentatin (DTN) from this plant using a bioassay-guided approach. DTN-induced cytotoxicity was observed with the MTT assay. Acridine orange/propidium iodide staining was used to detect cells in early apoptosis and high content screening (HCS) to observe nuclear condensation, cell permeability, mitochondrial membrane potential (MMP) and cytochrome c release. Apoptosis was confirmed with a clonogenic assay, DNA laddering and caspase 3/7 and 9 assays. Reactive oxygen species (ROS) formation, Bcl-2 and Bax expression, and cell cycle arrest were also investigated. The involvement of nuclear factor-kappa B (NF-κB) was analysed with the HCS assay.

Results

A significant increase in chromatin condensation in the cell nucleus was observed by fluorescence analysis. Apoptosis was confirmed by the reduced number of colonies in the clonogenic assay and the increased number of cellular DNA breaks in treated cells observed as a DNA ladder. Treatment of MCF-7 cells with DTN encouraged apoptosis with cell death-transducing signals that reduced MMP by down-regulation of Bcl-2 and up-regulation of Bax, triggering cytochrome c release from the mitochondria to the cytosol. The released cytochrome c triggered the activation of caspase 9 followed by the executioner caspase 3/7. DTN treatment significantly arrested MCF-7 cells at the G0/G1 phase (p<0.05) and ROS was significantly elevated. Moreover, DTN significantly blocked the induced translocation of NF-κB from cytoplasm to nucleus.

Conclusion

Together, the results demonstrated that the DTN isolated from Clausena excavata inhibited the proliferation of MCF-7 cells, leading to cell cycle arrest and programmed cell death, which was confirmed to occur through the mitochondrial pathway with involvement of the NF-κB signalling pathway.     

Effects of long-snake moxibustion on the HPA axis and liver 11β-HSD1 expression in rats with kidney yang deficiency: 长蛇灸对肾阳虚大鼠HPA轴及肝脏11β-HSD1的影响

ObjectiveTo observe the effects of long-snake moxibustion on the hypothalamic-pituitary-adrenocortical (HPA) axis and hepatic 11β-hydroxy steroid dehydrogenase type 1 (11β-HSD1) expression in rats with kidney yang deficiency to provide a basis for later in-depth exploration of the action mechanism of long-snake moxibustion on such rats.MethodsFifteen SPF-grade, male, SD rats were randomly divided into a blank control group, a model group, and a long-snake moxibustion treatment group, with five rats in each group. Hydrocortisone powder (30 mg/kg) was administered by gavage at a volume of 10 mL/kg to prepare the rat model of kidney yang deficiency. After successful modeling, the rats in the long-snake moxibustion treatment group underwent long-snake moxibustion treatment every other day (along the governor vessel from Dazhui (GV14) to Shenshu(BL23), for a period of 14 days. The remaining two groups were secured in the same way as the long-snake moxibustion treatment group, although they did not receive any treatment. The body weight, rectal temperature, and spontaneous activity count of the rats, as well as serum levels of corticotropin releasing hormone (CRH) and corticosterone (CORT) were detected by ELISA before modeling, after modeling, and after treatment. The amount of 11β-HSD1 protein in rat liver was determined by immunohistochemistry and Western blot analysis.ResultsCompared with the rats in the blank control group, those in the model group exhibited a significant decrease in the trend of body weight growth and in rectal temperature (P <0.05), as well as a slight yet non-significant decrease in spontaneous activity count (P >0.05); compared with the rats in the model group, the rats in the treatment group exhibited a significant increase in rectal temperature (P <0.05) and in spontaneous activity count (P <0.05). Moreover, after 14 days of treatment, compared with the rats in the blank, the rats in the model group exhibited a significant decrease in serum CORT content (P <0.05) and in the expression of 11β-HSD1 in the liver (P <0.05), as well as a slight yet non-significant decrease in serum CRH content (P >0.05); compared with the rats in the model group, the rats in the treatment group exhibited a significant increase in serum CRH content (P <0.05) and in the expression of 11β-HSD1 in the liver (P <0.05), as well as a slight yet non-significant increase in serum CORT content (P >0.05).ConclusionLong-snake moxibustion can increase the rectal temperature and spontaneous activity count of rats with kidney yang deficiency, improve the function of the HPA axis, and increase the expression of 11β-HSD1 in the liver.