Antrodin A from mycelium of Antrodia camphorata alleviates acute alcoholic liver injury and modulates intestinal flora dysbiosis in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Antrodia camphorata (A. camphorata) is a rare functional fungus in Taiwan and contains a variety of biologically active ingredients. Antrodin A (AdA) is one of the main active ingredients in the solid-state fermented A. camphorata mycelium. It protects the liver from alcohol damage by improving the antioxidant and anti-inflammatory capacity of the liver and maintaining the stability of the intestinal flora. AIM OF THE STUDY: The aim of this study was to evaluate the hepatoprotective activities of ethyl acetate layer extract (EALE), AdA, and Antroquinonol (Aq) from mycelium of A. camphorata on alcoholic liver injury. MATERIALS AND METHODS: Mice were given with intragastrically vehicle (NC, 2% CMC-Na), alcohol (AL, 12 mL/kg bw), or different A. camphorata samples (EALE, AdA, Aq) at low (100 mg/kg bw) or high (200 mg/kg bw) dosages. The positive control (PC) group was given with silymarin (200 mg/kg bw). Except the NC group, each group of mice was fasted for 4 h after the last treatment and was intragastrically administrated with 50% alcohol (12 mL/kg bw). At the end of experiment, mouse serum was collected and the liver was excised. A portion of the liver was fixed in formalin and used for histopathological analysis, whereas the rest was used for biochemical analysis and real-time PCR analysis. The intestinal flora structure of feces was analyzed by determining the v3-v4 region sequence in 16S rDNA. RESULTS: The high-dose groups of the three samples (EALEH, AdAH, and AqH) significantly alleviated the alcohol-induced increases in liver index, serum ALT, AST, and AKP activities. Serum TG level was significantly reduced in all treatment groups. The increase of HDL-C content indicated that active ingredients of A. camphorata could reduce the lipid content in serum. Furthermore, MDA contents of the AdAH and AqH groups in liver were significantly reduced, accompanying with the levels of SOD, CAT, and GSH elevated to various extents. Antioxidant and anti-inflammatory capabilities in the liver were increased in the AdAH group, as evidenced by the mRNA expression levels of Nrf-2 and HO-1 were significantly increased; while those of CYP2e1, TNF-α, and TLR-4 were significantly decreased. Analysis of intestinal flora of feces showed that alcohol treatment significantly changed the composition of intestinal flora. Supplementation with AdA could mitigate dysbiosis of intestinal flora induced by alcohol. Flora of Faecalibaculum, Lactobacillus, and Coriobacteriaceae_UCG-002 showed significantly negative correlations with ALT, AST, AKP, and MDA levels. CONCLUSION: Antrodin A could improve the antioxidant and anti-inflammatory capacities of the liver and maintain the stability of intestinal flora. It is potentially a good candidate compound against acute alcoholic liver injury.

[Comparison of integration processing technology of origin and traditional cutting processing technology of Moslae Herba for lung-Yang deficiency rats].

To compare the effect of integration processing technology of origin (IPTO) and traditional cutting processing technology (TCPT) of Moslae Herba for lung-Yang deficiency rats caused by complex factors, analyze the mechanism, and provide the modern pharmacology basis for the implementation of IPTO of Moslae Herba. The rat models of lung-Yang deficiency were established by smoking + swimming in ice water + drinking ice water. The model rats were randomly divided into different groups, and were treated with intragastric administration for 30 d. Then the general signs, anal temperature and autonomic activity of the rats were observed. The pathological morphology of lung tissues was observed, and the positive expression of tumor necrosis factor (TNF-α) was observed by immunohistochemical method, and the hematological indexes were determined. Enzyme linked immunosorbent assay (ELISA) method was used to detect serum nitric oxide (NO), immunoglobulin G (IGG), malondialdehyde (MDA), thromboxane B2 (TXB2) and interleukin-8 (IL-8) level, and the organ coefficients of heart, liver, spleen, lung, kidney and other organs were calculated. According to the results, Moslae Herba volatile oil and decoction could improve the general signs and autonomic activities of lung-Yang deficiency rats, improve the body weight, rectal temperature, and the content of IGG in serum of lung-Yang deficiency rats, reduce organ coefficients of heart, liver, spleen, lung and kidney, serum NO, MDA, TXB2, IL-8 contents, white blood cell and TNF-α mean optical density in the lung tissues of rats. witg statistically significant difference (P<0.01 or P<0.05). The effects of IPTO volatile oil and water decoction were slightly higher. Therefore, Moslae Herba has therapeutic effect on lung-Yang deficiency rats, and ICPT has better effect, whose mechanism may be related to the intervention of TNF-α expression, improving the level of IGG, and inhibiting NO, MDA, IL-8, and TXB2 levels.

Two new nortriterpenoid saponins from Salicornia bigelovii Torr. and their cytotoxic activity.

Investigation of characteristic constituents of Salicornia bigelovii Torr. led to isolation of two new 30-nortriterpenoid glycosides, Bigelovii A (1), Bigelovii B (2), together with two known 30-nortriterpenoid glycosides 3-4 and three known oleanane-type triterpenoid glycosides 5-7. The structures of new compounds were elucidated by extensive 1D and 2D NMR, and MS spectroscopic analysis, and chemical evidences. All compounds were isolated for the first time from Chenopodiaceae. Thus compounds 1-4 were evaluated for their cytotoxicity and compouds 1, 3 showed moderate activity against four cell lines, HL-60 (promyelocytic leukemia), MCF-7 (breast carcinoma), HepG2 (liver carcinoma) and A549 (lung carcinoma), with IC(50) values of 6.18, 78.08, 13.64 and >100µM for 1; 31.87, >100, ~100, >100µM for 3, respectively.

[Biotransformation of podophyllotoxin by cell suspension culture and root culture of Rheum palmatum].

OBJECTIVE: To study the biotransformation of podophyllotoxin by the cell suspension culture and root culture systems of Rheum palmatum. METHOD: Using plant tissue culture technology and HPLC techniques to isolate products. The structures were elucidated by spectroscopic means. RESULT: Cell suspension culture of R. palmatum could convert podophyllotoxin to produce picropodophyllotoxin with the yield of 73.8%, while root culture of R. palmatum could convert podophyllotoxin to produce epipodophyllotoxin and apopodophyllotoxin. CONCLUSION: Podophyllotoxin did not affect the pH value of the media used in tissue cultures. Both cell suspension culture and root culture of R. palmatum can convert podophyllotoxin.

[Induction of hairy roots and anthraquinone production in Rheum palmatum].

OBJECTIVE: To study the induction of hairy roots and anthraquinone production in Rheum palmatum. METHOD: Leaf blades, hypocotyls and petioles of R. palmatum were infected by Agrobacterium rhizogenes LBA9402 and A. rhizogenes R1601, respectively. RESULT: Hairy roots were induced by two strains of A. rhizogenes, and LBA9402 showed stronger infective ability than R1601. Three clones of hairy roots were cultured. DH7a was induced by R1601, DH5a and DH5c were induced by LBA9402. DH7a grew faster than DH5a and DH5c, and all of them grew faster obviously than normal root (NOR). There were significant differences in anthraquinone composition and content among four kinds of roots. Emodin physcion and chrysophanol were predominant anthraquinone in DH5a, DH5c and NOR respectivly. Aloe-emodin content was the lowest in all root cultures compared with other four anthraquinones. CONCLUSION: The in vitro culture system of the established hairy roots laid a foundation for mass production of anthraquinone by hairy root culture.

[Ri plasmid transformation of Glycyrrhiza uralensis and effects of some factors on growth of hairy roots].

OBJECTIVE: To study the induction of hairy roots of Glycyrrhiza uralensis and effect of some physical as well as chemical factors on the growth of hairy roots. METHOD: Hypocotyls and cotyledons of G. uralensis were infected with Agrobacterium rhizogenes LBA9402 and A. rhizogenes R1601, respectively. RESULT: Hairy roots were induced by two strains of A. rhizogenes, and LBA9402 showed astronger infective ability. The frequency of transformation of hypocotyls was higher than that of cotyledons. Among the different media tested, WP medium yielded the fastest growth rate of hairy roots. Light inhibited the growth of hairy roots. The total amout of 5 flavonoids in hairy roots was 1.5 times as much as that in the callus, among them licochalcone content was 15.5 times as that of callus. CONCLUSION: The in vitro culture system of the hairy roots established laid a foundation for mass production of flavonoids by hairy root culture.

[Effects of carbon source and nitrogen source on callus growth and flavonoid content in Glycyrrhiza uralensis].

OBJECTIVE: To study the effects of carbon source and nitrogen source on callus growth, and flavonoid content in Glycyrrhiza uralensis. METHOD: Induction and culture of callus were conducted in the media of different concentrations of sucrose, and fructose and ratio of NH4+/NO3- ,and flavonoid content was measured by HPLC. RESULT: The results showed that fructose was superior to sucrose for callus growth and flavonoid formation, and the optimum concentration was 2%. The flavonoids content was 2 times higher than that of sucrose as carbon sources. In the range of 2% -6% of fructose concentration, the flavonoid content was decreased along with the concentration of fructose, but the licochalcone was increased 5-fold. The highest flavonoid content of 151.47 microg x g(-1) was obtained when the ratio of NH4+/NO3- in the medium was 1/2. NH; inhibited the callus growth and flavonoid formation of G. uralensis. CONCLUSION: Fructose as carbon source was superior to sucrose for callus growth and flavonoid formation, and NO3- was favorable to the callus growth and flavonoids accumulation.