Micro-particulate Ganoderma lucidum spore β-glucanin enhances the antitumor activity of gemcitabin via remodeling immunosuppressive microenvironment in Lewis lung cancer / 药学学报

Gemcitabine (GEM) is a commonly used drug in the clinical treatment of non-small cell lung cancer. Due to the accumulation of cells mediating immune escape and T cell depletion after chemotherapy, tumor microenvironment (TME) tends to be immunosuppressive status, which ultimately leads to tumor metastasis. The experimental protocol was approved by the Medical Laboratory Animal Ethics Committee of Jiangsu Provincial Academy of Chinese Medicine. Therefore, we observed the immunomodulatory effects of micro-particulate Ganoderma lucidum spore β-glucan (PGSG) on macrophages in vitro experiments. Next, mice subcutaneous Lewis lung cancer models were established to observe the anti-tumor effects of PGSG through oral administration of PGSG combined with GEM. Flow cytometry analysis was used to analyze the ratio of anti-tumor T cells in tumors and spleen, as well as the proportion of myeloid-derived suppressor cells (MDSC), tumor-associated macrophages (TAM) and regulatory cells (Tregs). The results showed that PGSG can up-regulate the expression of major histocompatibility antigens (MHC-II), CD40, CD86 and CD80 on the surface of macrophages, enhance the ability to phagocytosis of neutral red and further mediate the release of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4) and interleukin-10 (IL-10). In vivo experiments, combined administration can significantly decrease the volume and weight of tumors, reduce the ratio of MDSC (CD11b+Gr-1+), M-MDSC (CD11b+Ly6G-Ly6Chigh) and Treg (CD4+Foxp3+). At the same time, PGSG promoted the conversion of M2 (F4/80+CD206+) to M1 (F4/80+MHC-II+) and enhanced the response of helper T cell-1 (Th1) (CD4+IFN-γ+) and cytotoxic T lymphocyte (CTL) (CD8+IFN-γ+), which is of great significance for killing tumors. These results suggest that PGSG can regulate innate and adaptive antitumor immune responses, reshape the immunosuppressive microenvironment and enhance the anti-lung cancer effect of GEM.

Effects and mechanism of epimedium polysaccharide on solubility of icariin and baohuoside Ⅰ / 中国中药杂志

This study evaluated the effects of epimedium polysaccharide(EPS) on the solubility of icariin and baohuoside Ⅰ so as to preliminary explore its solubilization function and the underlying mechanism. The solubility of these two insoluble flavonoids in water and polysaccharide solutions was compared by high performance liquid chromatography, and the mechanism was investigated by diffe-rential scanning calorimetry(DSC) and critical micelle concentration determination. The results indicated that their solubilization in crude EPS solutions was concentration-dependent. The solubility of icariin and baohuoside Ⅰ in 20 mg·mL~(-1) EPS-1-1 was 9.05 times and 5.76 times that in water, respectively; while their solubility in 20 mg·mL~(-1) EPS-2-1 was 10.55 and 8.39 times that in water, respectively. The change of the DSC thermograms suggested the formation of new complexes from icariin and baohuoside Ⅰ with polysaccharides. The critical micelle concentrations proved the micellar properties of both EPS-1-1 and EPS-2-1. In short, EPS can significantly increase the solubility of icariin and baohuoside Ⅰ, the mechanism of which may be related to the formation of micellar complexes between EPS and insoluble flavonoids.

Alkali extraction of Ganoderma lucidum β-glucan and its anti-tumor immune regulation / 药学学报

In this study, an immunostimulating particulate β-glucan was isolated from a hot alkaline extract of the fruiting bodies of Ganoderma lucidum. The optimum conditions of 8 hours treatment time, 1∶20 solid - liquid ratio and 55 ℃ for the alkaline extract process were obtained after investigating by single-factor experiments and Box-Benhnken design in terms of the Ganoderma lucidum particulate β-glucan (GLG) increment, and these conditions resulted in a GLG yield of 8.57%. The experimental protocol was approved by the Medical Laboratory Animal Ethics Committee of Jiangsu Provincial Academy of Chinese Medicine. The result showed that resident macrophages were effectively activated by GLG, such as with the up-regulation of co-stimulatory molecules, the secretion of cytokines and phagocytic uptake. GLG could also promote the proliferation of spleen lymphocytes in mice. In addition, IFN-γ production of spleen CD4+ T cells and cytotoxic T lymphocyte (CTL) responses were significantly enhanced on GLG orally treatment, which ultimately resulted in significantly decreased tumor burden. Taken together, these data suggest that GLG might act as an immune stimulator to exert antitumor effects.

Response surface analysis for the optimization of extraction condition for polysaccharides from Epimedium polysaccharides and studies on its tumor immune activities / 药学学报

To enhance the efficiency of the extraction for Epimedium koreanum Nakai polysaccharides, we conducted the Box-Benhnken experiment and examined the response surface. C57 BL6 mice were used as Lewis-bearing mice model in the study of the polysaccharides in the regulation of tumor immunity. In the first place, different factors including extraction time, temperature, and ratio were evaluated in the yield of polysaccharides. The second order polynomial equation was formed to fit the experimental data. The optimal condition was temperature 87℃, extraction time 3.84 h, and ratio of solution and material 1:16.33 to get polysaccharides from Epimedium koreanum Nakai. Under this condition, model-predicted and experimentally measured values of polysaccharide yield were 1.045% and 1.023%, respectively, with a relative error between them at 2.15%. The extraction method is reliable, simple and applicable to the extraction of Epimedium poly-saccharides. In addition, this data suggests that Epimedium polysaccharides have obvious immune-regulatory activities in the tumor-bearing mice through increasing the immune-enhancing cytokines to override the immune-suppressing factors.

Research thoughts on tumor immune responses by polysaccharide of Chinese medicine via oral administration / 中国中药杂志

Tumor immunotherapy is one of the most significant scientific progresses. The idea of applying the traditional Chinese theory of "the balance of Yin and Yang" to treat cancer is in accordance with that of modern tumor immune strategy. Researches indicated that polysaccharide of Chinese medicine through regulation in immune responses could offer better paradigm for tumor immune treatment under the traditional Chinese theory. However, current studies related to tumor immunotherapy largely focus on the immunity enhancement while lack of the exploration of suppressive factors. Meanwhile, the complex analysis and detection on composition as well as structure definitely increase the difficulty in mechanism of oral absorption and function in vivo. To better exploit novel Chinese medicine of polysaccharide for tumor immune treatment, this article will provide some constructive thoughts on regulation of tumor immune responses based on up to date researches of structure-function relationship, absorbent process and molecular mechanisms responsible for tumor immune as well.

Research thoughts on structural components of Chinese medicine combined with bioinformatics / 中国中药杂志

Traditional Chinese medicine(TCM) is a complex system, featured with integrity and characteristics. Structural component TCM is a well-organized integrity of traditional Chinese medicine, reflecting multi-component integration effect of TCM. It gives us a new view on the material basis of TCM. Currently, conventional researching strategies are not enough to deal with the relationship between material basis and efficacy, multi-composition, multi-targets, and multi-section mechanism. Post-genome area gives a birth to bioinformatics, which involves systematic biology, different levels of omics, corresponding mathematics and computer techniques. It increasingly becomes a powerful tool to understand complicated system and life essential laws. Research ideas, methods. and knowledge of data mining technology of bioinformatics combined with the theory of structural components of Chinese medicine bring a new opportunity for developing structural components of Chinese medicine, systematically exploring the essence of TCM and promoting the modernization of TCM.

Study on oral absorption enhancers of astragalus polysaccharides / 中国中药杂志

Astragalus polysaccharides was lounded to 4-(2-aminoethylphenol), followed by labeling the APS-Tyr with fluorescein-5-isothiocyanate (FITC) at the secondary amino group. The absorption enhancement effects of low molecular weight chitosan and protamine on astragalus polysaccharides were evaluated via Caco-2 cell culture model. The results show that the fluorecent labeling compound has good stability and high sensitivity. On the other hand low molecular weight chitosan and protamine also can promoted absorption of the astragalus polysaccharides without any cytotoxity, and the absorption increase was more significant with increasing the amount of low molecular weight chitosan and protamine. At the same time, the low molecular weight chitosan has slightly better effect. The transepithelial electric resistance (TEER) of Caco-2 cells show that absorption enhancers could improve its membrane transport permeability by opening tight junctions between cells and increasing the cell membrane fluidity.

Comparison of protective effects of eight ethyl acetate extracts from Eclipta prostrate on NHBE cells based on component structure theory / 中国中药杂志

To analyze and compare the protective effects of active components in different ethyl acetate extracts (EAEEPs) from Eclipta prostrate, in order to study the comparison of materials bases protecting normal human bronchial epithelial (NHBE) cells. The MTT assay was taken to compare the protective effect of different EAEEPs on cigarette smoke extracts (CSE) -induced NHBE cells. The ultra-performance liquid chromatography (UPLC) was applied to analyze the content of phenolic acid, coumaric grass ether and flavonoid in EAEEPs. According to the results, all of the eight EAEEPs (0-200 mg x L(-1)) showed certain protective effect on NHBE cells, with statistical difference. Specifically, the total mass of EAEEP VII (89.15 mg x L(-1)) and EAEEP VIII (57.44 mg x L(-1)), which showed the strongest activity, was not the highest, while EAEEP III (132.25 mg x L(-1)) displayed the highest total mass. In the combination with the "component structure" theory, the analysis showed a significant difference in the mass structure among phenolic acid, coumaric grass ether and flavonoid in EAEEP VIII and EAEEP VIII, which were 1.0: 1. 0: 0.5 and 1.0: 1.9: 0.8, respectively. The results suggested a specific optimal "component structure" relationship may exist in EAEEP, which could provide reference for the material base study and quality control.

Disposition of geniposide and genipin via intestinal absorption barrier / 中国药学杂志

OBJECTIVE: To study the disposition of geniposide and genipin via intestinal absorption barrier. METHODS: The biotransformation of geniposide was studied by incubating it with intestinal flora or intestinal enzymes. The intestinal absorption of genipin at duodenum, jejunum, ileum and colon was investigated using single-pass intestinal perfusion model. RESULTS: The metabolism activity of intestinal flora for geniposide was (1098.3±519.2) μmol · h-1 · g-1. The concentration of geniposide reduced from 20.00 to 9.60 μmol · L-1 after 4 h of incubation with intestinal enzymes, while the concentration of the metabolite of geniposide, genipin, increased to 3.52 μmol · L-1. The effective permeability coefficients(Peff*) of ginipin at duodenum, jejunum, ileum, and colon were 3.77±0.38, 3.00±0.20, 2.79±0.16, and 2.11 ±0.62, respectively, and the absorption percentages at different intestinal segments of 10 cm long(10 cm% ABS) were (70.24±7.88)%, (56.94 ±4.34)%, (53.44±3.73)%, and (48.52 ±9.59)%, respectively. There were significant differences between duodenum and the other regions of intestine of rats (P < 0.05). The phase II metabolite of genipin was found in rat bile, and its UV absorption spectrum was in accordance with that of genipin hydrolyzed by β-glu-curonidase. CONCLUSION: Geniposide can be transformed to genipin in rat intestine. Genipin can be well absorbed in general intestinal tract without specific absorption site. The phase II metabolite of genipin can be excreted to small intestine through bile.

Polybasic research on the biopharmaceutical characteristics of 20 (S)-protopanaxadiol / 药学学报

In this study, the biopharmaceutical properties of 20 (S)-protopanaxadiol (PPD) were studied. Firstly, the equilibrium solubility and apparent oil/water partition coefficient of PPD were used to predict the absorption in vivo. Meanwhile the membrane permeability and absorption window were studied by Caco-2 cell model and single-pass intestinal perfusion model. Furthermore, the bioavailability and metabolism were combined to study the absorption properties and metabolic properties in vivo. All of them were used to provide theoretical and practical foundation for designing PPD preparation. The results showed that PPD is poorly water-soluble, and the equilibrium solubility in water is only 35.24 mg x L(-1). The oil-water partition coefficient is 46.21 (logP = 1.66). By Caco-2 cell model, the results showed PPD uptake in general, and it also has efflux. By in situ intestinal perfusion model, the results showed that the absorption of PPD in the intestine is good, and the effective permeability coefficient were duodenum > jejunum > ileum > colon. The oral bioavailability of PPD was 29.39%. It was not well. Metabolic studies showed PPD in vivo presented a wide spread metabolism. So the main factors that restricted oral bioavailability of PPD were the poor solubility and first-pass effect.

Regulation mechanism of triterpenoid components from Prunella asiatica on phase II detoxifying enzymes in vitro and in vivo / 中国中药杂志

To study the effects of triterpenoid components from Prunella asiatica on phase II detoxifying enzymes and protein expression in vitro and in vivo. Normal human bronchial epithelial (NHBE) cell model was used in vitro, and the mouse model of Kunming (KM) mice was used in vivo. CDNB assay was used to measure the activity of GST. NADPH and DCIP was used to detect the activity of NQO1. DTNB colorimetric assay was used to detect GSH. Western blot was use to detect the protein expression of NQO1. We found that triterpenoid components from P. asiatica could increase the activity of GST, NQO1 and GSH in NHBE cells and KM mice. NQO1 protein expression can also be increased in vitro. The study suggests that triterpenoid components from P. asiatica can prevent the lung cancer by regulating the body phase II detoxification enzyme activity and protein expression.