Cytotoxic activity of sesquiterpenoids from Atractylodes ovata on leukemia cell lines

The rhizome of Atractylodes ovata (Bai Zhu in Chinese) is a widely used traditional Chinese herb in Taiwan as a tonic agent. In this paper, four sesquiterpenoids, namely atractylon, and atractylenolides I, II, and III, were isolated from the n-hexane extract of A. ovata and were evaluated for cytotoxic effects in vitro. Atractylon significantly inhibited the growth of human leukemia cell line HL-60 and mouse leukemia cell line P-388, and showed low cytotoxicity against primary cultures of normal human peripheral blood mononuclear cells at 15 microg/ml for 12 h. Atractylon had a dose-dependent antiproliferative effect on the two tumor cell lines. In accordance with DNA fragment increases and PARP protein decreases, atractylon at 15 microg/ml for 6 h induced apoptosis in HL-60 cells. Moreover, atractylon inhibited the viability of P-388 cells and induced apoptosis after 15 microg/ml treatment for 12 h in an in vitro assay. However, atractylenolide I at 30 microg/ml for 12 h also induced apoptosis in HL-60 and P-388 cells, but atractylenolides II and III showed no significant inhibition effects on tumor cell growth. As the above results suggested, atractylon and atractylenolide I were the major cytotoxic principle constituents of A. ovata on leukemia cell lines.     

Atractylenolide III reduces NLRP3 inflammasome activation and Th1/Th2 imbalances in both in vitro and in vivo models of asthma

Paediatric asthma is a common inflammatory disease in children. Atractylenolide III is an active component of the Atractylodes rhizome, an herbal medicine that has been used as an asthma treatment. This study aimed to explore the effects and underlying mechanisms of atractylenolide III in IL-4-induced 16HBE cells and ovalbumin-induced asthmatic mice. The results showed that IL-4 stimulation significantly decreased, and atractylenolide III treatment increased, growth and apoptosis of 16HBE cells. In 16HBE cells, administration of atractylenolide III also significantly suppressed the IL-4-induced increases in the expression of cleaved caspase-1; apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC); and nucleotide-binding domain and leucine-rich repeat protein 3 (NLRP3). Moreover, the numbers of total leukocytes, neutrophils, eosinophils, and macrophages significantly increased in ovalbumin-induced mice, and then decreased after atractylenolide III treatment. In ovalbumin-induced asthmatic mice, atractylenolide III treatment also significantly inhibited NLRP3 inflammasome activation and restored the Th1/Th2 balance. These results indicate that atractylenolide III reduced NLRP3 inflammasome activation and regulated the Th1/Th2 balance in IL-4 induced 16HBE cells and ovalbumin-induced asthmatic mice, suggesting it has a protective effect that may be useful in the treatment of paediatric asthma.     

Preventive effects of a fractioned polysaccharide from a traditional Chinese herbal medical formula (Yu Ping Feng San) on carbon tetrachloride‐induced hepatic fibrosis

Objectives The study was to investigate the prevention effects and possible mechanism of Yu Ping Feng San fractioned polysaccharide (YPF‐P) on CCl₄‐induced liver fibrosis in rats. Methods YPF‐P was prepared from root of Astragalus membranaceus, rhizome of Atractylodes macrocephaia and root of Raidix saposhnikoviae, and compared with polysaccharide from root of Astragalus membranaceus (AP). Hepatic fibrosis was induced by subcutaneous injection with carbon tetrachloride twice weekly for 12 weeks in Sprague–Dawley rats. YPF‐P, AP and colchicine were administered intragastrically daily to carbon tetrachloride‐treated rats. Histopathological changes of the liver and hepatic stellate cells were evaluated by Masson staining and transmission electron microscopy, respectively. Markers of fibrosis were determined by radioimmunoassay, biochemistry assay and ELISA. The mRNA expressions of tissue inhibitor of metalloproteinase‐1 (TIMP‐1), matrix metalloproteinase‐13 (MMP‐13), procollagen I and collagen III were detected by RT‐PCR. Key findings YPF‐P dose‐dependently alleviated the degree of liver fibrosis and inhibited hepatic stellate cell transformation into myofibroblast‐like cells, markedly reduced the elevated levels of hyaluronic acid, laminin, type IV collagen, type III procollagen, hydroxyproline and transforming growth factor beta‐1, suppressed procollagen I, collagen III and TIMP‐1 expression, and improved the TIMP‐1/MMP‐13 ratio. MMP‐13 expression was only promoted moderately by YPF‐P. Compared with AP, YPF‐P showed more potency on most markers except laminin, type IV collagen and MMP‐13 mRNA. Conclusions YPF‐P prevented the progress of rat liver fibrosis induced by carbon tetrachloride and had a more potent preventative effect. The preventative effect may be associated with the ability of YPF‐P to inhibit the synthesis of matrix collagen and balance the TIMP/MMP system.     

白术内酯Ⅰ对人胃癌细胞SGC-7901裸鼠移植瘤生长及凋亡的影响

目的：研究白术内酯Ⅰ（atractylenolide Ⅰ）对人胃癌细胞SGC-7901裸鼠移植瘤生长及凋亡相关蛋白Bax、cleaved caspase-3、p53、Bcl-2表达的影响。方法：建立SGC-7901裸鼠移植瘤模型,观察白术内酯Ⅰ对肿瘤生长的影响;TUNEL法检测移植瘤组织中的细胞凋亡;Western blotting检测瘤组织中Bax、Bcl-2、cleaved caspase-3及p53蛋白表达。结果：白术内酯Ⅰ不同程度抑制裸鼠SGC-7901移植瘤的生长,与对照组比较,给药后肿瘤体积（TV,tumor volume）、相对肿瘤体积（RTV,relative tumor volume）和相对肿瘤增殖率[T/C（%）,T_RTV/C_RTV]明显下降;移植瘤组织中凋亡细胞明显增多;白术内酯Ⅰ上调移植瘤组织中Bax、cleaved caspase-3及p53的蛋白表达,下调Bcl-2 的蛋白表达。结论：白术内酯Ⅰ能明显抑制人胃癌细胞SGC-7901裸鼠移植瘤的生长,分子机制主要包括增加Bax、cleaved caspase-3、p53蛋白表达,减少Bcl-2蛋白表达,最终导致肿瘤细胞凋亡。     

Role of Blepharis maderaspatensis and Ammannia baccifera plant extracts on in vitro oxygen radical scavenging,secretion of gastric fluid and gastroprotection on ulcer induced rats

Context: Blepharis maderaspatensis L. Roth (BM) (Acanthaceae) and Ammannia baccifera L. (AB) (Lythraceae) are used in folk medicine for various stomach disorders.

Objective: The chloroform and ethanol extracts of both plants were evaluated for antioxidant, gastric antisecretory, and gastroprotective properties.

Methods: Antioxidant properties of the extracts were evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay and nitric oxide (NO) scavenging assay. The gastric antisecretory properties of the extracts were assessed, at a dose of 100 and 200 mg/kg, using aspirin-pylorus ligation induced gastric ulcer models and the gastroprotective activity of the extracts was assessed, at a dose of 100 and 200 mg/kg, using HCl-ethanol induced ulcer models in rats.

Results and discussion: Ethanol extract of BM (EBM) possessed good antioxidant property with IC₅₀ values of 37.4 and 44.1 µg/mL in DPPH and NO scavenging assays respectively, where 25–250 µg/mL concentration in DPPH assay and 30–300 µg/mL concentration in NO scavenging assay were used. Ethanol extract of AB (EAB) at a dose of 200 mg/kg reduced the free acidity to 142.66 mEq/L and total acidity to 451.22 mEq/l. It reduced the gastric secretion with increase in pH from 2.2 to 3.15. Possessing good antisecretory activity, it also reduced the ulcer by 92.2% in aspirin and pylorus ligation induced gastric ulcer models. EAB increased the mucus secretion and adherent mucus in the tissues with a 71.43% reduction of ulcerin HCl-ethanol induced ulcer models, at a dose of 200 mg/kg. This activity can be attributed to the various flavonoids like rutin and kaempferol-3-O-β-glucopyranoside, and the phytosterol, β-sitosterol-3-O-β-glucopyranoside, and phenolics present in the extracts.

Conclusion: EBM possessed significant antioxidant property while EAB possessed good antisecretory and gastroprotective activity.     

Effects of the rhizomes of Atractylodes japonica and atractylenolide I on allergic response and experimental atopic dermatitis

Although some anti-allergic activities of the rhizome of Atractylodes japonica have been previously reported, the active principle(s) for anti-allergic action is not fully elucidated and the effect of this plant material on atopic dermatitis (AD) is not known. In this study, the 70% ethanol extract of the rhizome of A. japonica was found to significantly inhibit 5-lipoxygenase (5-LOX)-catalyzed leukotrienes (LT) production from rat basophilic leukemia (RBL)-1 cells. From the extract of A. japonica, three major sesquiterpene derivatives including atractylenolide I, atractylenolide III and eudesma-4,7-dien-8-one were successfully isolated. Among these compounds, only atractylenolide I was shown to strongly inhibit 5-LOX from RBL-1 cells (IC₅₀ = 18.6 ??M). To evaluate the effects of experimental AD, the ethanol extract of A. japonica (200 mg/day) was administered orally to hapten-treated NC/Nga mice which is an animal model of AD. It was firstly found that the extract significantly inhibited AD-like symptoms in mice, as judged by severity score and scratching behavior. Taken together, it is concluded that A. japonica possesses the inhibitory activity on 5-LOX and an animal model of AD, and atractylenolide I may contribute, at least in part, to these anti-allergic actions of A. japonica.     

Activity of Brucea javanica oil emulsion against gastric ulcers in rodents

The present study aims to investigate the gastroprotective effect of Brucea javanica oil emulsion (BJOE) in animals. Gastroprotective potential of BJOE was studied on absolute ethanol, aspirin, reserpine and restraint plus water immersion-induced gastric ulcers in mice as well as glacial acetic acid (GAA) and pyloric ligation (PL)-induced gastric ulcers in rats. Except for ulcer scores, total acidity as well as pepsin activity as for the PL-induced gastric ulcer model and ulcer incidence as for the GAA-induced gastric ulcer model were also determined. Histopathological evaluation as for aspirin, reserpine, PL-induced models was conducted. Results showed that BJOE significantly (P < 0.05) reduced ulcer index in the mouse and rat models in a dose-dependent manner. It had significant (P < 0.05) suppressive effect on total activity of gastric juice as well in PL-induced model. Histopathological examination for the stomach samples confirmed the findings in the aspirin, reserpine or PL-induced gastric lesion models, which showed relatively complete mucosa structure and less inflammation. It is concluded that BJOE could be effective on gastric ulcer in rodents and its gastroprotective activity might be related to antioxidant, anti-inflammatory ability and promote gastric mucus secreted. The results may provide beneficial basis for increasing BJOE's clinical indication in future.     

Anti-ulcer actions of phytosphingosine hydrochloride in different experimental rat ulcer models

The gastroprotective activity of phytosphingosine hydrochloride (PS-HCl, CAS 554-62-1) was assessed in four different rat models of experimentally induced gastric ulcer. Various doses (2.5-10 mg/kg) of PS-HCI were orally administered to rats 30 min before the treatment with HCl/ethanol, indometacin, cysteamine, or to rats with ligated pylorus. Oral administration of PS-HCl (2.5-10 mg/kg) to rats prevented the acute ulcer formation in 4 different types of ulcer in a dose-dependent manner as follows: (1) HCl/ethanol-induced gastric mucosal membrane lesions (20.1-47.8% inhibition), (2) indometacin-induced gastric mucosal membrane lesions (4.6-31.9% inhibition), (3) duodenal ulcer induced by cysteamine (10-20% inhibition), (4) gastric secretion and ulceration following pylorus ligation (33.3-61.9% inhibition). These results indicate that PS-HCI may be useful for the prevention of gastric ulcer.     

Gastroprotective activity of α-terpineol in two experimental models of gastric ulcer in rats

BACKGROUND AND THE PURPOSE OF THE STUDY: Several plant essential oils, as well as terpenes present in essential oils, have shown gastroprotective activity. The aim of the present work was to evaluate the gastroprotective activity of α-terpineol, a monoterpene alcohol which is present in essential oils of various plants. METHODS: The gastroprotective activity of α-terpineol was evaluated in rats by assessing the changes in ethanol and indomethacin-induced gastric ulcer scores and on gastric secretory volume and total acidity in pylorus-ligated rats. Alpha-terpineol was administrated orally at the doses of 10, 30, and 50 mg/kg one hour before administration of the ulcer inducing agents by the pylorus ligation procedure. The involvement of endogenous prostaglandins in the protective effect of α-terpineol in ethanol-induced gastric lesions test was assessed by administration of indomethacin (10 mg/kg, s.c.) 30 min before oral administration of α-terpineol at the dose of 50 mg/kg. RESULTS: α-terpineol presented gastroprotective activity against ethanol-induced ulcers at the doses of 10, 30, and 50 mg/kg. Epoxy-carvone at the dose of 10 mg/kg did not present gastroprotective activity against ulcer induced by indomethacin, but at the doses of 30 and 50 mg/kg it attenuated the gastric damages induced by this agent significantly. Pretreatment with indomethacin did not prevent the gastroprotective effect of α-terpineol on ethanol-induced ulcers. Alpha-terpineol also did not affect the gastric secretion in pylorus-ligated rats. MAJOR CONCLUSION: The results suggest that α-terpineol presents gastroprotective action which does not involve either an increase in the synthesis of endogenous prostaglandin or a decrease in the gastric acid secretion.     

Synthesis and Biological Evaluation of 3, 8‐dimethyl‐5‐isopropylazulene Derivatives as Anti‐gastric Ulcer Agent

Recent studies showed that Guaiazulene (GA) and Sodium guaiazulene sulfonate (GAS‐Na) have good anti‐gastric ulcer effect. Here, two series of GA derivatives were synthesized and evaluated for their anti‐gastric ulcer activity. The data obtained from in vivo testing of these compounds in a rodent ethanol‐induced stomach injury model are discussed. Among the tested compounds, A1 , A4 , and A9 (ulcer index: 1.125 ± 1.246**, 1.714 ± 0.756*, 1.875 ± 1.126*) exhibited better anti‐gastric ulcer activity than the positive control Omeprazole (2.005 ± 1.011*). The information got from these studies and the results of 3D‐SAR investigation may be useful in the design of novel anti‐gastric ulcer agents.     

Neuroinflammatory reactions in experimental gastric ulcer: Target for mucosal protection

The effect of different opioids receptor agonists—morphine, DAGO (μ-agonists), DADLE, DPDPE and deltorphin II (δ-agonists)—on gastric mucosal damage induced by either acidified ethanol or acidified aspirin was studied following subcutaneous (sc) administration of these agonists. The results indicate that both μ and δ receptors are involved in gastroprotection. Morphine, DAGO and DADLE, injected intracerebroventricularly, were also effective in both ulcer models. This suggests that gastric cytoprotection can be induced also be central action, since gastric acid secretion is not involved in the pathomechanism of mucosal damage induced by acidified ethanol. Interaction between the opioids and α₂-adrenoceptors in gastroprotection is suggested by the findings that the gastroprotective effect of clonidine (0.09 μmol/kg orally) was antagonized by opioid antagonists. As both naloxone (1.38 μmol/kg sc) and naltrindole (12 μmol/kg sc) exerted antagonist effects, both μ and δ receptors are likely to be involved in presynaptic α₂-receptor-mediated gastroprotection.     

Gastroprotective and antisecretory effects of <Emphasis Type="Italic">Ailanthus excelsa</Emphasis> (Roxb)

Ailanthus excelsa (Roxb), an Egyptian medicinal species highly important for treating numerous diseases, was investigated against experimentally induced gastric ulcer in rodents. We evaluated the gastroprotective effect of four extracts (petroleum ether, diethyl ether, chloroform, and methanol) of A. excelsa bark by using the ethanol-induced gastric lesion model. The pretreatment of animals with methanolic, petroleum ether, and chloroformic extracts (100 mg/kg, oral (p.o.)) from A. excelsa significantly reduced gastric lesion induced by ulcerogenic agent (56, 47, and 70%, respectively) when compared with animals pretreated with vehicle. However, the diethyl ether pretreatment led to the least gastric lesion damage (83%), similar to the standard antiulcer drug, cimetidine, at the same dose (100 mg/kg, p.o.). The lower effective dose of diethyl ether extract, as well as cimetidine, given by intraduodenal route, significantly increased the pH values and reduced the acid output of gastric juice. Sterols, triterpenes, and quassinoids are present in the diethyl ether extract of A. excelsa stem bark, which presented the best gastroprotective action among the studied extracts. Our study confirmed the traditional indications of A. excelsa for the treatment of gastric ulcer.     

Neuroprotection by herbal formula FBD and its active compounds

FBD, a Chinese herbal formula, composed of Fu Ling [the sclerotium of the fungus of Poria cocos (Schw.) Wolf (Polyporaceae)], Bai Zhu [the rhizome of Atractylodes macrocephala Koidz.(Compositae)], and Danggui [the root of Angelica sinensis (Oliv.) Diels (Umbelliferae)], has been found beneficial in treating brain ischemia-reperfusion (I/R). In this work, oral pretreatment with supercritical CO₂ extract (FBD-CO₂, 37.?5?mg/kg) twice daily for 3.5 days, significantly attenuated brain infarction (p < 0.05), blocked neuron specific enolase (NSE) efflux (p < 0.05) in mice subjected to repetitive 10?min of common carotid artery occlusion following 24?h reperfusion, whether coupled with aqueous extract (FBD-H₂O, 150?mg/kg) or not. Except atractylenolide I and pachymic acid, atractylenolide II, III, levistolid A, and ferulic acid isolated from FBD-CO₂ alone protected neuron-like PC12 cells obviously and concentration-dependently against I/R-like insults (p < 0.05 ~ 0.01) induced by sodium dithionite (10 mM), monosodium glutamate (2 mM), KCl (0.2?M), or hydrogen peroxide (0.2 mM) at 1-100 μM. Even though at sub-active concentrations (< 1 μM), the combination of the six components contained in FBD-CO₂ (10 μg/mL), protected markedly PC12 cells (p < 0.01), in a synergistic fashion. Moreover, intraperitoneal treatment with dual doses of 37.?5?mg/kg FBD-CO₂ and 1.?2?mg/kg combination reduced both infarct size (p < 0.01 and p > 0.05, respectively) and NSE efflux (p < 0.05 and p > 0.05, respectively). Therefore, neuroprotection by FBD on I/R induced neuronal injury originated partially from the synergies of its compounds atractylenolide II, atractylenolide III, levistolid A and ferulic acid.     

Effect of <Emphasis Type="Italic">Gastrodia elata</Emphasis> on tumor necrosis factor-alpha-induced matrix metalloproteinase activity in endothelial cells

The aim of the present study was to investigate whether an ethanol extract of Gastrodia elata (EGE) rhizome, a traditional Korean herbal medical food, suppresses the endothelial extracellular matrix degradation induced by tumor necrosis factor (TNF)-α. Gelatin zymography results showed that pretreatment with EGE to human umbilical vein endothelial cells (HUVEC) decreased TNF-α-induced increase of matrix metalloproteinase (MMP)-2/-9 activities in the range of 1–50 μg/ml. Real-time qRT-PCR results also revealed that TNF-α-induced MMP-2/-9 mRNA expression levels were attenuated by pretreatment with EGE. These results provide new insights into the pathophysiological mechanisms for the anti-atherosclerotic properties of EGE in vascular diseases.     

Ethanol consumption increases the expression of endothelial nitric oxide synthase, inducible nitric oxide synthase and metalloproteinases in the rat kidney

Objectives The effects of longterm ethanol consumption on the levels of nitric oxide (NO) and the expression of endothelial NO synthase (eNOS), inducible NO synthase (iNOS) and metalloproteinase‐2 (MMP‐2) were studied in rat kidney. Methods Male Wistar rats were treated with 20% ethanol (v/v) for 6 weeks. Nitrite and nitrate generation was measured by chemiluminescence. Protein and mRNA levels of eNOS and iNOS were assessed by immunohistochemistry and quantitative real‐time polymerase chain reaction, respectively. MMP‐2 activity was determined by gelatin zymography. Histopathological changes in kidneys and indices of renal function (creatinine and urea) and tissue injury (mitochondrial respiration) were also investigated. Results Chronic ethanol consumption did not alter malondialdehyde levels in the kidney. Ethanol consumption induced a significant increase in renal nitrite and nitrate levels. Treatment with ethanol increased mRNA expression of both eNOS and iNOS. Immunohistochemical assays showed increased immunostaining for eNOS and iNOS after treatment with ethanol. Kidneys from ethanol‐treated rats showed increased activity of MMP‐2. Histopathological investigation of kidneys from ethanol‐treated animals revealed tubular necrosis. Indices of renal function and tissue injury were not altered in ethanol‐treated rats. Conclusions Ethanol consumption increased renal metalloproteinase expression/activity, which was accompanied by histopathological changes in the kidney and elevated NO generation. Since iNOS‐derived NO and MMPs contribute to progressive renal injury, the increased levels of NO and MMPs observed in ethanol‐treated rats might contribute to progressive renal damage.     

Two new compounds from Atractylodes macrocephala with neuroprotective activity

In the present study, two new compounds, together with six known compounds, were isolated from rhizome of Atractylodes macrocephala Koidz by a series of silica gel, ODS column chromatography, and preparative HPLC. Their structures were characterized as atractylenolide II (1), atractylenolide I (2), biepiasterolid (3), isoatractylenolide I (4), atractylenolide III (5), 3β-acetoxyl atractylenolide I (6), (4E,6E,12E)-tetradeca-4,6,12-triene-8,10-diyne-13,14-triol (7), (3S,4E,6E,12E)-1-acetoxy-tetradeca-4,6,12-triene-8,10-diyne-3,14-diol (8) on the basis of 1D, 2D NMR, and circular dichroism analyses. Among them, compounds 6 and 8 were novel compounds. In addition, their neuroprotective activity against MPP⁺-induced SH-SY5Y cells was evaluated by MTT colorimetry. The results showed that all these compounds have definite protective effect on MPP⁺-induced SH-SY5Y cells.     

The antioxidative and antihistaminic effect of Nigella sativa and its major constituent, thymoquinone on ethanol-induced gastric mucosal damage

The aim of this study was to assess the possible protective effects of Nigella sativa (NS) and its constituent, thymoquinone (TQ) on ethanol-induced gastric mucosal damage in an experimental model. Forty male rats aged four months were divided into four groups (each group containing ten animals); the control group received physiologic saline (10 ml kg⁻¹) and the ethanol group had taken 1 ml (per rat) absolute alcohol by gavage. The third and fourth groups also received NS (500 mg kg⁻¹) and TQ (10 mg kg⁻¹) by gavage 1 h before alcohol administration, respectively. Both drugs (NS and TQ) could protect the gastric mucosa against the injurious effect of absolute alcohol and promote ulcer healing as evidenced from the ulcer index values. Gastric damage was confirmed histomorphometrically by significant increases in the number of mast cells (MC) and gastric erosions in ethanol treated rats. The NS treatment significantly decreased the number of MC and reduced the area of gastric erosions. Likewise, TQ treatment was also able to reduce the number of MC and the gravity of gastric mucosal lesions, but to lesser extent compared to NS. Gastric tissue histamine levels and myeloperoxidase activities were found to be increased in ethanol treated rats, and NS or TQ treatment reversed these increases. Results obtained from this study suggest that both drugs, particularly NS could partly protect gastric mucosa from acute alcohol-induced mucosal injury, and these gastroprotective effects could be due to their antiperoxidative, antioxidant and antihistaminic effects.