Uncovering the pharmacological mechanism of Shou Tai Wan on recurrent spontaneous abortion: A integrated pharmacology strategy-based research.

ETHNOPHARMACOLOGICAL RELEVANCE: Shou Tai Wan (STW), a traditional Chinese medicine formula, has been historically used for the treatment of recurrent spontaneous abortion (RSA). Despite its long-standing usage, the exact mechanism underlying the therapeutic effects of STW remains unclear in the existing literature. AIMS OF THIS STUDY: To explore the Pharmacological Mechanism of STW on RSA. METHODS: A network pharmacological methodology was utilized to predict the active compounds and potential targets of STW, collect the RSA targets and other human proteins of STW, and analyze the STW related networks. The animal experiments were also performed to validate the effect of STW on RSA. RESULTS: The results of network analysis showed that STW may regulate PI3K/AKT, MAPK, FoxO signaling pathways and so on. Animal experiment established the RSA model with CBA/J × DBA/2 mice. It was found that STW can reduce the embryo absorption rate of RSA group (p < 0.05) and balance the expression of Th 1/Th2 type cytokines compared with the model group. After 14 days of administration, the decidual and placental tissues were taken and the CD4+ T cells were isolated, and the phosphorylation level of signaling pathway was detected by Springbio720 antibody microarray. This experiment found that STW can significantly up-regulate the phosphorylation levels of STAT3 and STAT6 proteins in the STAT signaling pathway, and down-regulating the phosphorylation level of STAT1 protein. STW also significantly up-regulated the phosphorylation levels of Raf1, A-Raf, Ask1, Mek1, Mek2, JKK1, ERK1, ERK2, c-fos, c-Jun and CREB proteins in the MAPK signaling pathway, and down-regulate the phosphorylation levels of MEK6 and IKKb proteins. Compared with the RSA group, the STW group increased the expression levels of ERK1/2 mRNA and proteins and p-ERK1/2 proteins, and there was a statistical difference (p < 0.05). This is consistent with the chip results. CONCLUSION: STW may achieve therapeutic effects by interfering with the signaling pathways, biological processes and targets discovered in this study. It provides a new perspective for revealing the immunological mechanism of STW in the treatment of RSA, and also provides a theoretical basis for the clinical use of STW in the treatment of RSA.

Structure analysis of a non-esterified homogalacturonan isolated from Portulaca oleracea L. and its adjuvant effect in OVA-immunized mice.

We isolated and purified a pectin from Portulaca oleracea L. (P. oleracea), and analysed its structure by high-performance size exclusion chromatography (HPSEC), high-performance liquid chromatography (HPLC), gas chromatograph-mass spectrometer (GC-MS), fourier transform infrared spectroscopy (FT-IR), and 1H, 13C nuclear magnetic resonance spectroscopy (NMR). The data indicated that this pectin (designated as POPW-HG) was a linear non-esterified homogalacturonan, which is unique in plants; its molecular weight was around 41.2 kDa. Meanwhile, POPW-HG as an adjuvant was evaluated in the mice immunized with OVA subcutaneously. OVA-specific antibody titres from the sera of immunized mice were tested by ELISA. It showed that POPW-HG significantly enhanced OVA-specific antibody titres (IgG, IgG1, and IgG2b) (p < 0.05) in a dose-dependent manner in the OVA-immunized mice, preliminarily indicating POPW-HG could increase an antibody response, Th1 and Th2 immune response. In addition, the ratio of IgG1/IgG2b suggested POPW-HG induced a Th2-biased response in the OVA-immunized mice. The results demonstrated POPW-HG could be a potential adjuvant candidate in vaccines.

Water-soluble polysaccharide isolated with alkali from the stem of Physalis alkekengi L.: structural characterization and immunologic enhancement in DNA vaccine.

A water-soluble polysaccharide (WSPA) was isolated with alkali and purified from the mature stem of Physalis alkekengi L. WSPA (Mw=31kDa) was an acid heteropolysaccharide, which consisted of Rha, Ara, Xyl, Gal, Glc and GalA in ratio of 1.0:2.5:0.8:2.7:4.4:1.4. The results from structural analysis indicated backbone and branches of WSPA were composed of (1→3)-linked Glc, (1→3)-linked Gal, (1→2)-linked Xyl, (1→2)-linked Ara, and (1→2)-linked Rha residues. However, GalA was distributed only in the backbone of WSPA. All branches of WSPA were at O-2 of (1→6)-linked Gal and terminated with Glc. More importantly, it was found that WSPA significantly enhanced specific antibody IgG response with higher titers of IgG1 as well as IgG2b (p<0.05) in mice immunized with DNA vaccine. Therefore, WSPA can be considered as a potential adjuvant candidate in DNA vaccine.

Adjuvant effect of polysaccharide from fruits of Physalis alkekengi L. in DNA vaccine against systemic candidiasis.

Adjuvant effect mediated by polysaccharide (PPSB) isolated from the fruits of Physalis alkekengi L. in DNA vaccine was evaluated in mice. Recombinant plasmid containing epitope C (LKVIRK) from heat shock protein 90 (HSP90) of Candida albicans (C. albican) was used as DNA vaccine (pD-HSP90C). The results indicated that PPSB significantly enhanced specific antibody titers IgG, IgG1, IgG2b, and concentration of IL-2 and IL-4 in sera of mice immunized with pD-HSP90C (p<0.05). More importantly, it was found that the mice immunized with pD-HSP90C/PPSB not only had fewer CFU (colony forming unites) in the kidneys than mice immunized with pD-HSP90C, but also a statistically significant higher survival rate over PBS-injected group (p<0.05) when the immunized mice were challenged with living C. albican cells. However, no statistically significant difference in survival rate was observed between pD-HSP90C-immunized group and PBS-injected group. Therefore, PPSB can be considered as a promising adjuvant eliciting both Th1 and Th2 responses to enhance the efficacy of DNA vaccines.

Characterization of the multiple absorbed constituents in rats after oral administration of Chai-Huang decoction by liquid chromatography coupled with electrospray-ionization mass spectrometry.

A rapid, sensitive, and specific method by high-performance liquid chromatography (HPLC) coupled to diode-array detection (DAD) and tandem mass spectrometry (MS) techniques was developed for the identification of absorbed constituents and their metabolites in rats after the oral administration of a Chai-Huang decoction (CHD), which consists of Bupleurum chinense and Scutellaria baicalensis in the proportion 1 : 1 (w/w). By comparing their retention times and MS data with those of authentic compounds and published data, a total of 14 compounds were identified in the CHD samples. In addition, eleven and seven compounds were characterized in the urine and serum samples of the rats, respectively. The results indicated that the main absorbed constituents were chrysin-6-C-arabinosyl-8-C-glucoside, chrysin-6-C-glucosyl-8-C-arabinoside, baicalin, wogonin-5-O-glucoside, oroxylin A-7-O-glucuronide, wogonoside, saikosaponin A, saikosaponin C, saikosaponin D, baicalein, and wogonin. These compounds might be responsible for the curative effects of the CHD. The findings demonstrated that the proposed method could be used to rapidly and simultaneously analyze and screen the multiple absorbed bioactive constituents in a formula of traditional Chinese medicines (TCM). This is very important not only for the pharmaceutical discovery process and the quality control of crude drugs but also to explain the mechanisms of action of TCM.

[Studies on phenolic compounds from Sophora alopecuroides].

OBJECTIVE: To study the chemical constituents of Sophora alopecuroides. METHOD: The non-alkaloid constituents on 95% ethanol extracts of S. alopecuroides were separated and purified with various chromatographic method after solvent extraction and their structures were elucidated by means of physical property and the analysis of their spectral data. RESULT: Seven compounds were isolated and identified as ferulic acid (1), butein (2), 7,3',4'-trihydroxyursaflavonoid (3), 7-hydroxy-3', 4'-methylenedioxyisoflavone (4), butein4-O-beta-D-glucopyranoside (5), piscidic acid (6) and 7,3',4'-trihydroxyur-sadimethoxyflavan-7-O-beta-D-glucopyranoside (7). CONCLUSION: Seven compounds were obtained from this plant for the first time.