樟芝多糖通过降低NLRP3 Caspase1炎性小体表达改善帕金森小鼠神经行为学的机制研究

目的:研究樟芝多糖通过降低NLRP3-Caspase1炎性小体表达改善6-OHDA构建的帕金森小鼠模型的行为学机制。方法:利用6-OHDA脑内注射构建帕金森小鼠模型，通过酪氨酸羟化酶（TH）免疫组化染色和行为学判定小鼠模型的构建成功。利用樟芝多糖进行干预，分别在干预前、干预后的第1、3、7天4个时间点进行神经行为学实验，分别采用转棒实验、爬杆实验检测小鼠自主行为能力以及协调能力，4个时间点取小鼠尾静脉外周血采用ELISA法检测外周血中Caspase1和IL-1β的表达，樟芝多糖干预第7天时待进行完行为学实验后小鼠断颈处死，取小鼠脑组织-纹状体，Western blot法检测纹状体中Caspase1、proCaspase1、NLRP3的表达，高效液相色谱检测纹状体中单胺类神经递质的表达，RT-QPCR检测Caspase1、NLRP3、IL-1β、IL-4、IL-6的mRNA表达。NISSl染色检测小鼠脑组织神经细胞凋亡情况。 结果:6-OHDA脑内注射可以造成小鼠帕金森样病变，且TH蛋白表达显著下调，樟芝多糖干预后小鼠的行为学得到显著改善（P<0.05），纹状体中Caspase1、proCaspase1、NLRP3的表达显著下调，与模型小鼠相比具有统计学差异（P<0.05），且相关炎症因子Caspase1、NLRP3、IL-1β、IL-4、IL-6的mRNA表达下调（P<0.05），纹状体中单胺类神经递质表达上升（P<0.05）。结论:樟芝多糖可以通过下调NLRP3-Caspase1炎性小体表达来改善6-OHDA构建的帕金森小鼠模型行为改善，这可能是樟芝多糖治疗帕金森的机制之一。     

4-Acetylantroquinonol B Suppresses Prostate Cancer Growth and Angiogenesis via a VEGF/PI3K/ERK/mTOR-Dependent Signaling Pathway in Subcutaneous Xenograft and In Vivo Angiogenesis Models

Prostate cancer is a major cause of cancer-related mortality in men in developed countries. The compound, 4-acetylantroquinonol B (4AAQB), is isolated from Antrodia cinnamomea (commonly known as Niu-Chang-Chih), which has been shown to inhibit cancer growth. However, the anticancer activity of 4AAQB has not previously been examined in prostate cancer. This study aimed to investigate the effect of 4AAQB on cancer and angiogenesis, as well as to explore its mechanism of action. Human prostate cancer cells (PC3) and human umbilical vein endothelial cells (HUVEC) were used in cell viability, cell migration, and cell cycle functional assays to evaluate the anticancer and antiangiogenic efficacy of 4AAQB in vitro. The effects of 4AAQB in vivo were determined using xenograft and angiogenesis models. The signaling events downstream of 4AAQB were also examined. The 4AAQB compound inhibited PC3 cell growth and migration, and reduced in vivo cancer growth, as shown in a subcutaneous xenograft model. Furthermore, 4AAQB inhibited HUVEC migration, tube formation, and aortic ring sprouting; it also reduced neovascularization in a Matrigel implant angiogenesis assay in vivo. The 4AAQB compound also decreased metastasis in the PC3 prostate cancer model in vivo. Serum or vascular endothelial growth factor (VEGF)-induced VEGF receptor 2 (VEGFR2), phosphoinositide 3-kinase (PI3K)/Ak strain transforming (Akt), and extracellular signal-regulated kinase ½ (ERK ½) phosphorylation were attenuated by 4AAQB in both PC3 and HUVEC. In conclusion, 4AAQB is a potential candidate for prostate cancer therapy.     

樟芝真菌发酵培养基优化研究

以麸皮、蛋白胨、MgSO4&#183;7H2O、VB1、葡萄糖、酵母膏、KH2PO4。7种为影响樟芝真菌液体培养的影响因素,分别以菌丝体和胞外多糖为目的产物进行L18（3^7）正交试验,综合以胞外多糖和菌丝体为目的产物进行樟芝真菌液体培养的正交试验分析后得出,樟芝真菌液体培养时的适宜培养基为：麸皮40g／L、葡萄糖25g／L、蛋白胨4g／L、MgSO4&#183;7H2O 0．4g／L、酵母膏2．0g／L、KH2PO4 0．6g／L、VB10．01g／L。     

Antioxidant properties of methanolic extracts from Antrodia camphorata with various doses of γ-irradiation

The mycelia of Antrodia camphorata (Chang & Chou) Wu, Ryvarden & Chang were irradiated with gamma rays at doses of 2.5, 5, 10, 15 and 20 kGy and the antioxidant properties of the methanolic extracts were studied. At 2.5 mg/ml, antioxidant activities of methanolic extracts from 10 to 20 kGy-irradiated mycelia were significantly higher than those of the non-irradiated control. Reducing powers of methanolic extracts from unirradiated and 0.5–7.5 kGy-irradiated mycelia were comparable except for the 20 kGy-irradiated mycelia. At 2.5 mg/ml, all methanolic extracts showed excellent scavenging abilities of 92.3–103% on DPPH radicals. Scavenging abilities of methanolic extracts from 2.5 to 20 kGy irradiated mycelia were better than that of the unirradiated control at 10 mg/ml. With irradiation at 5–20 kGy, mycelia possessed higher chelating ability on ferrous ions than did the unirradiated control. The EC₅₀ values were below 15 mg/ml, except for values of scavenging ability of the unirradiated control on hydroxyl radicals. Total phenols were the major naturally occurring antioxidant components found in the range of 13.0–15.5 mg/g. In summary, γ-irradiation not only maintained the antioxidant properties of mycelia but also enhanced the antioxidant properties to some extent.     

Inhibitory effect of Antrodia camphorata constituents on the Helicobacter pylori-associated gastric inflammation

Helicobacter pylori infection plays a crucial role in the pathogenesis of peptic ulcer, and gastric cancer. Among the 13 chemical constituents isolated from fruiting bodies of Antrodia camphorata, methyl antcinate B (4), antcins K (10) and A (12) displayed potential anti-H. pylori activity with inhibition zones of 13, 12 and 10 mm, respectively, at a concentration of 0.2 mM. The isolates 4 and 10 exhibited a dose response inhibition of H. pylori adhesion and invasion to AGS cells in a range of concentrations between 0.005 and 0.02 mM, while 12 has moderate effect at relatively higher concentration. Furthermore, at these concentrations (0.005–0.02 mM) the isolates 4 and 10 also inhibited the H. pylori-induced nuclear factor (NF)-кB activation, and the subsequent release of interleukin (IL)-8 in AGS cells. These results open the possibility of considering A. camphorata a chemopreventive agent for peptic ulcer or gastric cancer, but this bioactivity should be confirmed in vivo in the future.     

The 4‐acetylantroquinonol B isolated from mycelium of Antrodia cinnamomea inhibits proliferation of hepatoma cells

BACKGROUND: Antrodia cinnamomea is known for its antihepatoma activity, yet the identity of its active compound was unclear. In this study, a 5‐ton fermenter was used to prepare sufficient mycelium of A. cinnamomea for active compound isolation and identification. RESULTS: Using antiproliferative activity toward HepG2 cells as guidance in the isolation process, 4‐acetylantroquinonol B was purified and identified to be the major bioactive compound of A. cinnamomea cultivated by submerged fermentation. The median effective doses (EC₅₀) of 4‐acetylantroquinonol B for HepG2 cells were 0.10 ± 0.00 and 0.08 ± 0.00 µg mL^?1 for 72 and 96 h treatments, respectively. The selective indices of 4‐acetylantroquinonol B were 100 and 125 for 72 and 96 h treatments, respectively, indicating that this compound had high selective activity for hepatoma cells. CONCLUSION: 4‐Acetylantroquinonol B is the major antihepatoma constituent of Antrodia cinnamomea mycelium produced by submerged fermentation. Copyright © 2010 Society of Chemical Industry     

Ethanolic extracts of Antrodia cinnamomea mycelia fermented at varied times and scales have differential effects on hepatoma cells and normal primary hepatocytes

Mycelia of Antrodia cinnamomea (AC), an edible fungus native to Taiwan, were produced by submerged fermentation with various fermentation times in 250 mL, 5 and 500 L fermentors and were evaluated for the effect of fermentation products on the viabilities of Hep3B and HepG2 hepatoma cells and normal primary rat hepatocytes. The results showed that the ethanolic extracts of AC mycelia (from 250 mL fermentation for 8 wk and 5 and 500 L fermentations for 4 wk) possessed high antihepatoma activity. The IC(50) of ethanolic extract of AC mycelia fermented for 8 wk in a 250 mL fermentor against Hep3B and HepG2 cells were 82.9 and 54.2 microg/mL, respectively. Furthermore, the IC(50) for Hep3B and HepG2, treated with ethanolic extract of AC mycelia fermented for 4 wk in the 5 L fermentor were 48.7 and 3.8 microg/mL, respectively. Those treated with ethanolic extract of AC mycelia fermented for 4 wk in the 500 L fermentor were 36.9 and 3.1 microg/mL, respectively. No adverse effects of all samples on normal primary rat hepatocytes were observed.     

响应面法优化樟芝固态发酵产安卓奎诺尔

以樟芝菌固态发酵生产活性代谢产物安卓奎诺尔为目标物,采用Box-Behnken原理进行响应面分析,对樟芝固态发酵产安卓奎诺尔条件进行优化,结果表明：接种量为296.80 mL/kg、Triton X-100添加量为1.10 mL/kg、辅酶Q0的添加量为0.23 g/kg时,理论上樟芝固态发酵培养基安卓奎诺尔产量的最大值为865.85 mg/kg。经验证,安卓奎诺尔实际产量为865.32 mg/kg,表明实验建立的模型能较好地预测实际发酵产安卓奎诺尔的情况。通过优化,樟芝固态发酵安卓奎诺尔产量比优化前（260.57 mg/kg）提高了232.09%。     

An efficient total synthesis of a potent anti-inflammatory agent, benzocamphorin f, and its anti-inflammatory activity

A naturally occurring enynyl-benzenoid, benzocamphorin F (1), from the edible fungus Taiwanofungus camphoratus (Antrodia camphorata) was characterized by comprehensive spectral analysis. It displays anti-inflammatory bioactivity and is valuable for further biological studies. The present study is the first total synthesis of benzocamphorin F and the developed strategy described is a more efficient procedure that allowe the large-scale production of benzocamphorin F for further research of the biological activity both in vitro and in vivo.     

樟芝提取物Fr.2组分对PDGF-BB活化CFSC-8B的抑制作用

为了探讨樟芝分离组分Fr.2对血小板衍生生长因子-BB(PDGF-BB)刺激的大鼠肝星状细胞(HSCs)的作用,建立PDGF-BB活化的CFSC-8B细胞模型,并对细胞增殖、迁移、细胞外基质表达及机制进行了研究。结果表明,10 ng/mL PDGF-BB作用48 h可诱导CFSC-8B细胞增殖;Fr.2的IC50值为56.46μg/mL;25μg/mL Fr.2显著抑制PDGF-BB刺激CFSC-8B细胞的增殖及其迁移,下调α-SMA、Collagen I(Col 1)、CollagenⅢ(Col 3)和Fibronectin(Fn)mRNA表达水平,并下调p-ERK、p-JNK、p-P38蛋白质表达。表明樟芝Fr.2可抑制PDGF-BB诱导的细胞增殖、迁移及细胞外基质的表达,其机制可能与抑制MAPK信号转导通路有关。