2,3,5,4'- tetrahydroxystilbene-2-O-ß-D-glycoside biosynthesis by suspension cells cultures of Polygonum multiflorum Thunb and production enhancement by methyl jasmonate and salicylic acid.

Friable calli of Polygonum multiflorum Thunb have been induced in MS medium supplemented with 6-benzylaminopurine (6-BA) and kinetin (KT). Suspension cultures were initiated from friable calli by inoculating calli in liquid MS medium in shake flasks in the dark and 25 °C on an orbital shaker at 100 rpm. The maximum dry weight (DW, 7.85 g/L) and 2,3,5,4'-tetrahydroxystilbene-2-O-ß-D-glycoside (THSG, 56.39 mg/L) of suspension cells was obtained in MS medium after 16 days culture. Both methyl jasmonate (MeJA) and salicylic acid (SA) could increase THSG production. The most appropriate concentration of MeJA was 100 µmol/L in MS medium, in which concentration THSG content reached the maximum value of 147.79 mg/L, which represented a 162.36% increase compared to that of the control (56.33 mg/L). The most appropriate concentration of SA was 125 µmol/L in MS medium, at which concentration THSG content reached its maximum value of 116.43 mg/L, a 106.69% increase compared to that of the control (56.33 mg/L).

[Microscopic and polariscopic characteristics of 30 medicinal plants of Polygonum].

Polygonum L. s. str., belonging to Polygonaceae family, is a big genus with abundant medicinal plants. More than 10 plants are specified in Chinese Pharmacopoeia and many local medicinal standards and over 50 species are used as folk medicines. Owing to the similar morphologies and very small flowers and fruits, they are uneasily identified and often confused with each other and misused clinically. In order to provide a basis for identification of Polygonum s. str. plants, a histological study on stems and leaves of 30 species from Polygonum was undertaken by a routine/polarized light microscopy for the first time. The results showed that: (1) the transverse sections of stems of Polygonum are relatively similar, sclerenchyma such as xylem and fibres with strong polarization effects; (2) the surface views of leaves of Polygonum are distinguishable on distributions and types of stomata, with or without attachments (such as glandular hairs/scales or non-glandular hairs) and the polariscopic features of epidermal cell walls, stomata and cell contents. Observed under polarized light, it was found for the first time that stomata on leaf surface of some plants have a Maltese-cross effect with the arms of the cross intersecting at the stomatal opening. As a result, a key combining the microscopic and polariscopic characteristics of the stems as well as leaves was provided for identifying the 30 medicinal plants of Polygonum. The polarized light microscopic method was proven to be one of the quick, simple and effective techniques for the identification of medicinal plants and botanic crude materials.

Stimulative effects of Polygonum amplexicaule var. sinense on osteoblastic MC3T3-E1 cells.

CONTEXT: Polygonum amplexicaule D. Don var. sinense Forb. (Polygonaceae) (PAF) is a well known traditional herb used to treat some diseases, such as fractures, rheumatoid arthritis, muscle injury, and pain. However, its pharmacological mechanism of promoting the healing of fractures is still unknown. OBJECTIVE: The present study was designed to investigate the effects of PAF ethanol extracts on the proliferation and differentiation of osteoblastic MC3T3-E1 cell in vitro, thereby to illuminate the pharmacological mechanism to promote the healing of fractures. MATERIALS AND METHODS: The effects of PAF ethanol extracts on MC3T3-E1 cell proliferation and differentiation were detected by using CCK-8, cell cycle, alkaline phosphatase (ALP), and prostaglandin E(2) (PGE(2)) assays in vitro. RESULTS: The results showed that PAF ethanol extracts significantly stimulated cell proliferation at 0.1-100 µg/mL and the proportion of cells in S-phase increased from 16.33 to 27.29% in osteoblastic MC3T3-E1 cells. Moreover, PAF ethanol extracts increased ALP expression in MC3T3-E1 cells at the concentration from 0.1 to 100 µg/mL and inhibited PGE(2) production induced by TNF-α in osteoblasts at the concentrations ranging from 10 to 100 µg/mL in MC3T3-E1 osteoblasts. DISCUSSION AND CONCLUSION: These results indicated that PAF directly stimulates cell proliferation and differentiation of osteoblasts; therefore, this study preliminarily explored the pharmacological mechanism of PAF to promote the healing of bone rheumatism and various fractures.

Simultaneous identification and quantification of anthraquinones, polydatin, and resveratrol in Polygonum multiflorum, various Polygonum species, and dietary supplements by liquid chromatography and microscopic study of Polygonum species.

A high-performance liquid chromatographic (HPLC) method with ultraviolet absorption detection was developed to determine the presence of anthraquinones, polydatin, and resveratrol in Polygonum multiflorum Thunb. as well as other medicinal Polygonum species, viz., P. cuspidatum, P. oriental, P. aviculare, and P. vulgare, as well as commercial products that claim to contain P. multiflorum. The best results were obtained with a Phenomenex Gemini C18 column using gradient mobile phase composed of water (0.1% acetic acid) and acetonitrile (0.1% acetic acid). Elution was at a flow rate of 1.0 mL/min. The detection wavelength was 280 nm for anthraquinones and 320 nm for polydatin and resveratrol. The main anthraquinones identified were emodin and physcion. The HPLC pattern of P. multiflorum was also compared with 5 other species of Polygonum. The method was validated for precision, repeatability, and accuracy. The relative standard deviation was between 0.9 and 1.6%. The method was sensitive, quick, and accurate for determination of anthraquinones, polydatin, and resveratrol in 6 different species of Polygonum and can be used for quality control of P. multiflorum. The commercial samples and the 6 Polygonum species were compared microscopically, and a detailed description is provided for P. multiflorum.

[Pharmacognostical identification of Polygonum perfoliatum].

In the paper, phamacognostical identification and UV-identification of Polygonum perfoliatum L. were studied.

[Pharmacognostical identification on Chinese drug zhizhuliao].

In this paper, 2 kinds of Zhizhuliao that original plants are Polygonum suffultum Maxim., P. amplexicaule D. Don. var. sinense Forb. et Hemal. are made an initial study on the characteristics, microscopical and physichemical sides. It provides scientific basis for its comprehensive development and ultilization, and to use in clinical.

[Pharmacognostical identification on the Chinese drug quanshen].

In this paper, 5 kinds of Quanshen that original plants are Polygonum bistorta L., P. viviparum L., P. paleacum Wall., P. sphaerostachyum Meisn., P. macrophyllum D. Don. are made an initial study on the characteristics, microscopical and physichemistical sides. It provide scientific basis for its comprehensive development and ultilization, and to use in clinical.

Histochemical analysis of the root tuber of Polygonum multiflorum Thunb. (Fam. Polygonaceae).

Authentication is the first priority in quality evaluation of Chinese herbal medicines (CHMs). The most commonly used authentication methods are morphological identification and microscopic identification. Unfortunately, these two methods cannot provide the chemical information needed to assess the quality of CHMs. In this study, a combination of fluorescence microscopy and high performance liquid chromatography-mass spectrometry (HPLC-MS) was used to analyze the chemical profiles of the tissues of the raw root tubers of Polygonum multiflorum Thunb. The results showed that the cork cells, cortex, and vessels in transverse sections of the raw root tuber of P. multiflorum fluoresced differently. Further analysis by HPLC-MS revealed that anthraquinones are mainly distributed in the cortex, and 2,3,5,4'-tetrahydroxystilbene-2-O-ß-D-glucopyranoside could be found in all tissues of the raw root tubers of P. multiflorum with its relative content as cork > cortex > xylem of allotype vascular bundles > xylem of central vascular bundles. Moreover, the fluorescence characteristics of the tissues from the steamed root tuber of P. multiflorum were compared and showed different fluorescence from those of raw material. From these results, it can be deduced that the root tuber of P. multiflorum with a broader cortex and fewer vascular bundles visible in a transverse section should be of better quality. The different fluorescence characteristics can be used to differentiate raw root tubers of P. multiflorum from those that have been steamed.

[Histochemical localization and quantitative analysis of anthraquinones in rhizome of Polygonum cuspidatum].

Anthraquinones were histochemically locayed and content were determined in rhizome of Polygonum cuspidatum. Anthraquinones were located in the parenchyma cells of rhizome, including vascular ray, part of parenchyma cell in phloem, cortex and pith. Phelloderm and ray initial also accumulated a small amount anthraquinones. The content of total anthraquinones was highest in pith that consisted of parenchyma cells, and higher in bark (including phloem, cortex and periderm) than in xylem. The content of total anthraquinones was highest in three-year-old rhizome, and higher in biennial rhizome than in annual rhizome. It suggests that correlation exist between accumulation of anthraquinones and the growth age of rhizome.