Inhibitory effects of triterpenes isolated from Hoelen on free radical-induced lysis of red blood cells

Hoelen, sclederma of Poria cocos Wolf, has long been used as a sedative and diuretic in traditional medicine. Formerly, we demonstrated that Hoelen in vitro protects red blood cells from AAPH-induced hemolysis. In this study, tests were carried out to identify the main ingredient of Hoelen that has the scavenging effect on free-radicals. Triterpene carboxylic acids isolated from the methanol extract of Hoelen, i.e. pachymic acid, polyporenic acid, 3-epidehydrotumulosic acid, 3beta-hydroxylanosta-7,9(11), 24-trien-21-oic acid and 3-o-acetyl-16 alpha -hydroxytrametenolic acid, were found to have inhibitory activities against AAPH-induced lysis of red blood cells.     

Inhibitory effects of Oren-Gedoku-To (Huanglian-Jie-Du-Tang) on free radical-induced lysis of human red blood cells

Oren-gedoku-to (Huanglian-Jie-Du-Tang, OGT) has been used for the treatment of cerebrovascular disease, hypertension, gastritis and liver disease in Japan. The present study was to test our hypothesis that ingestion of Oren-gedoku-to extract (TJ-15) would protect red blood cell (RBC) membrane from free radical-induced oxidation if antioxidants in OGT could be absorbed and circulated in blood. When incubated with RBC suspension, OGT and its four constituting herbs provided strong protection for RBC membrane to hemolysis induced by 2,2-azo-bis (2-amidinopropane) dihydrochloride (AAPH), an azo free radical initiator. The inhibitory effect was in a dose-dependent manner at concentrations of 5 microgram/ml to 500 microgram/ml. Furthermore, the ingestion of 7.5 g of OGT (daily dose) was associated with a significant decrease in susceptibility of RBC to hemolysis in humans. The direct protection of RBC membrane from free-radical attack as observed in the present study could provide an important pathophysiological basis for making use of the favorable hemorheological effect of OGT.     

Protective effects of Terminalia arjuna against Doxorubicin-induced cardiotoxicity

Terminalia arjuna has been marked as a potential cardioprotective agent since vedic period. The present study was aimed to investigate the effects of butanolic fraction of Terminalia arjuna bark (TA-05) on Doxorubicin (Dox)-induced cardiotoxicity. Male wistar rats were used as in vivo model for the study. TA-05 was administered orally to Wistar rats at different doses (0.42 mg/kg, 0.85 mg/kg, 1.7 mg/kg, 3.4 mg/kg and 6.8 mg/kg) for 6 days/week for 4 weeks. Thereafter, all the animals except saline and TA-05-treated controls were administered 20 mg/kg Dox intraperitonially. There was a significant decrease in myocardial superoxide dismutase (38.94%) and reduced glutathione (23.84%) in animals treated with Dox. Concurrently marked increase in serum creatine kinase-MB (CKMB) activity (48.11%) as well as increase in extent of lipid peroxidation (2.55-fold) was reported. Co-treatment of TA-05 and Dox resulted in an increase in the cardiac antioxidant enzymes, decrease in serum CKMB levels and reduction in lipid peroxidation as compared to Dox-treated animals. Electron microscopic studies in Dox-treated animals revealed mitochondrial swelling, Z-band disarray, focal dilatation of smooth endoplasmic reticulum (SER) and lipid inclusions, whereas the concurrent administration of TA-05 led to a lesser degree of Dox-induced histological alterations. These findings suggest that butanolic fraction of Terminalia arjuna bark has protective effects against Dox-induced cardiotoxicity and may have potential as a cardioprotective agent.     

Inhibitory effects of Keishi-bukuryo-gan on free radical induced lysis of rat red blood cells

Keishi-bukuryo-gan (KBG) prevents the progression of atherosclerosis in cholesterol-fed rabbits by its antioxidative effect. The present study was to test our hypothesis that ingestion of KBG would protect red blood cell (RBC) membranes from free radical induced oxidation if polyphenolic antioxidants in KBG could be absorbed and circulated in the blood. When incubated with a RBC suspension, KBG and four of five herb medicines constituting KBG provided strong protection for RBC membranes against haemolysis induced by 2,2-azo-bis (2-amidinopropane) dihydrochloride (AAPH), an azo free radical initiator. The inhibitory effect was dose dependent at concentrations of 100-1000 microg/mL. Furthermore, the ingestion of 200 mg of KBG was associated with a significant decrease in susceptibility of RBC to haemolysis in rats.     

Antioxidant activity of Antrodia camphorata on free radical-induced endothelial cell damage

AIM OF THE STUDY: Antrodia camphorata (A. camphorata) is well known in Taiwan as a traditional Chinese medicine. The purpose of this study is to evaluate the antioxidant activity of Antrodia camphorata on free radical-induced endothelial cell damage. MATERIALS AND METHODS: In this study, a human umbilical vein endothelial cell (EC) culture system was used to evaluate the effects of the fermented culture broth of A. camphorata (FCBA) and aqueous extracts of mycelia from A. camphorata (AEMA) against the oxidative cell damage induced by the free-radical generator AAPH. RESULTS: The present investigations show that FCBA (25-100 microg/mL) and AEMA (50-200 microg/mL) effectively protect the ECs from damage after exposure to 15 mM AAPH for 16h. However, cell viability was not affected in ECs under controlled conditions after FCBA or AEMA treatment. An increase in EC prostacyclin (PGI(2)) production in response to AAPH exposure was positively and negatively correlated with cell damage and FCBA/AEMA concentration, respectively. Both FCBA and AEMA treatment significantly inhibited AAPH-apoptotic cell death in the ECs, as evidence by reduced DNA fragmentation, cytochrome c release, caspase-3 activation, and dysregulation of Bcl-2 and Bax. Moreover, the AAPH-induced reductions in EC SOD activity and protein levels are prevented by FCBA and AEMA. CONCLUSION: Our findings suggest that A. camphorata possesses antioxidant properties and improves endothelial function, further offering effective protection from atherosclerosis.     

马齿苋总黄酮对氧自由基引发人红细胞膜损伤的保护作用

 目的研究马齿苋总黄酮对超氧阴离子(O₂^-)引发的人红细胞膜氧化损伤的保护作用。方法 采用邻苯三酚自氧化产生的O₂^-引发人红细胞膜氧化损伤模型,研究马齿苋总黄酮对红细胞膜氧化损伤的影响。结果O₂^-能引起红细胞膜脂质过氧化,丙二醛(MDA)含量、自氧化速率显著增加,膜脂流动性及膜封闭度下降。而红细胞膜经一定量马齿苋总黄酮(60,120,240,480μg·mL^-1)预先处理后,膜MDA含量明显减少,自氧化速率明显降低,膜脂流动性和膜封闭度显著增高。结论马齿苋对氧自由基引发的人红细胞膜氧化损伤有一定的保护作用。     

Protective effect of Apocynum venetum (罗布麻) against lipid peroxidation damage of erythrocyte membrane

Objective: To investigate the protective effect of Apocynum venetum on lipid peroxidation damage of erythrocyte membrane. Methods: Model of lipid peroxidation of erythrocyte membrane was manufactured by three kinds of radicals generation systems. To set up a normal control group (NC), a model control group (MC), and four Apocynum venetum groups(A.V). Observation was made on the content of malondialdehyde (MDA). Results: Compared with MC group, all of the four Apocynum venetum groups dose-dependently inhibited the increases of MDA content in the membrane induced by xanthine - xanthine oxidase system, H2O2 or UV light. Conclusions: Apocynum venetum may protect erythrocyte membrane from the lipid peroxidative damage induced by radicals.     

川芎嗪对大鼠心肌缺血损伤的拮抗作用

目的 :研究川芎嗪对异丙肾上腺素致大鼠心肌缺血损伤的保护作用。方法 :采用皮下注射异丙肾上腺素 (ISP ,5mg·kg-1) ,2 4h后再注射同剂量一次 ,造成心肌缺血损伤模型。观测心肌线粒体中Ca2 + ATP酶、Ca2 + Mg2 + ATP酶活力以及Ca2 + 含量的变化。采用免疫组化法检测Bcl 2和Bax蛋白水平的变化。结果 :川芎嗪保护组心肌线粒体Ca2 + ATP酶、Ca2 + Mg2 + ATP酶活力较缺血组均有显著性升高 (P <0 .0 1) ,川芎嗪可稳定心肌线粒体Ca2 + 含量 ,增加缺血心肌组织中Bcl 2的表达 (P <0 .0 1) ,对Bax的表达影响不大 (P >0 .0 5 )。结论 :川芎嗪对大鼠心肌缺血损伤具有保护作用 ,该作用可能与提高心肌线粒体Ca2 + ATP酶、Ca2 + Mg2 + ATP酶活力及调控Bcl 2基因的表达有关。     

Protective effects of lycopene and tomato extract against doxorubicin-induced cardiotoxicity

The protective effect of tomato extract and lycopene on acute doxorubicin (DOX) myocardial toxicity was evaluated in mice. DOX toxicity, induced by a single intraperitoneal injection (15 mg/kg), was revealed by an elevated serum CPK(MB) and histopathological observations. Tomato extract (1.2 and 2.4 g/kg, i.p.) and lycopene (1.7 and 3.5 mg/kg, i.p.) prevented the rise in serum CPK(MB) and ameliorated cardiac cell injury. These results suggest that tomato extract and lycopene inhibit DOX cardiotoxicity and might serve as a novel combination chemotherapeutic agent with DOX to limit free radical-mediated organ injury.     

Protective effects of paeoniflorin against corticosterone-induced neurotoxicity in PC12 cells

Neuroprotection has been proposed as one of the acting mechanisms of antidepressants. Paeoniflorin, a monoterpene glycoside, has been reported to display antidepressant-like effects in animal models of behavioural despair. The present study aimed to examine the protective effect of paeoniflorin treatment on corticosterone-induced neurotoxicity in cultured rat pheochromocytoma (PC12) cells. Paeoniflorin was shown to elevate cell viability, decrease levels of intracellular reactive oxygen species (ROS) and malondialdehyde (MDA) in corticosterone-treated PC12 cells. Paeoniflorin also reversed the reduced nerve growth factor (NGF) mRNA level caused by corticosterone in PC12 cells. The results suggest that paeoniflorin exerts a neuroprotective effect on corticosterone-induced neurotoxicity in PC12 cells, at least in part, via the inhibition of oxidative stress and the up-regulation of NGF expression. This neuroprotective effect may be one of the action pathways that accounts for the in vivo antidepressant activity of paeoniflorin.     

Protective effects of hyperoside against carbon tetrachloride-induced liver damage in mice

In this study, the hepatoprotective effects of hyperoside (1), a flavonoid glycoside isolated from Artemisia capillaris, have been examined against carbon tetrachloride (CCl4)-induced liver injury. Mice were treated intraperitoneally with vehicle or 1 (50, 100, and 200 mg·kg(-1)) 30 min before and 2 h after CCl4 (20 μL·kg(-1)) injection. Levels of serum aminotransferases were increased 24 h after CCl4 injection, and these increases were attenuated by 1. Histological analysis showed that 1 prevented portal inflammation, centrizonal necrosis, and Kupffer cell hyperplasia. Lipid peroxidation was increased and hepatic glutathione content was decreased significantly after CCl4 treatment, and these changes were reduced by administration of 1. Protein and mRNA expression of tumor necrosis factor-α (TNF-α), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and heme oxygenase-1 (HO-1) and nuclear protein expression of nuclear factor erythroid 2-related factor 2 (Nrf2) significantly increased after CCl4 injection. Compound 1 suppressed TNF-α, iNOS, and COX-2 protein and mRNA expression and augmented HO-1 protein and mRNA expression and Nrf2 nuclear protein expression. These results suggest that 1 has protective effects against CCl4-induced acute liver injury, and this protection is likely due to enhancement of the antioxidative defense system and suppression of the inflammatory response.     

Protective effects of HV-P411 complex against D-galactosamine-induced hepatotoxicity in rats

This study examined the hepatoprotective effect of the HV-P411 complex, an herbal extract mixture from the seeds of Vitis vinifera, Schisandra chinensis and Taraxacum officinale, against D-galactosamine (D-GalN)-induced hepatitis. Hepatotoxicity was induced by D-GalN (700 mg/kg, i.p.), and the HV-P411 complex was administered orally 48, 24, and 2 h before and 6 h after D-GalN injection. Increases in serum aminotransferase activity and lipid peroxidation and a decrease in hepatic glutathione content were attenuated by the HV-P411 complex 24 h after D-GalN treatment. The HV-P411 complex attenuated the increases in serum tumor necrosis factor-α, interleukin (IL)-6 level and cyclooxygenase-2 protein production and their mRNA expressions, while increases in serum IL-10 level and heme oxygenase-1 protein production and their mRNA expressions were augmented by the HV-P411 complex. The increased translocation of nuclear factor-κB and c-Jun phosphorylation were attenuated by treatment with the HV-P411 complex. Our results suggest that the HV-P411 complex prevents D-GalN-induced hepatotoxicity via antioxidative and anti-inflammatory activities.     

Protective effects of curcumin against liver fibrosis through modulating DNA methylation

Recent research has demonstrated that advanced liver fibrosis in patients could be reversed, but no approved agents are available for the treatment and prevention of liver fibrosis in humans. Curcumin(CUR) is the principal curcuminoid of turmeric. Inhibitory effects of CUR and its underlying mechanisms in liver fibrogenesis have been explored. In the present study, we hypothesized that epigenetic mechanisms contribute to the protective effects of CUR against liver fibrosis. We used CCl4-induced liver injury in BALB/c mice and the rat hepatic stellate cell line HSC-T6 as experimental models. Genomic DNA methylation was analyzed by Me DIP-chip and verified by real-time PCR on Me DIP-enriched DNA. The m RNA and protein expressions of DNMT1, α-SMA, and Col1α1 were determined by real-time PCR and Western blotting, respectively. The methylation statuses of FGFR3, FZD10, Gpx4, and Hoxd3 were further confirmed by quantitative methylation-specific PCR(q MSP). Our results showed that CUR treatment reversed liver injury in vivo and in vitro, possibly through down regulation of DNMT1, α-SMA, and Col1α1 and by demethylation of the key genes. In conclusion, aberrant methylation is closely associated with liver fibrosis and CUR treatment may reverse liver fibrosis by epigenetic mechanisms.     

金丝桃苷对大鼠心肌缺血再灌注损伤的保护作用

目的 探讨金丝桃苷（Hyp)对大鼠心肌缺血后再灌注导致的损伤的保护作用及作用的机制。方法 大鼠心肌缺血30min后再灌3.5h造成心肌缺血再灌注损伤模型；测定造模前腹腔注射Hyp对损伤大鼠心电图和血清中CPK及心肌组织中MDA，SOD和NO生成的影响。观察损伤大鼠心肌细胞凋亡的形成和Hyp干预的结果。结果 Hyp可以改变缺血再灌注损伤大鼠心电图T波的变化幅度，减少缺血再灌注损伤导致的大鼠心律失常发生率，抑制大鼠血清CPK的程式高和心肌组织中MDA，NO的形成，提高SOD的活力；抑制损伤大鼠心肌细胞凋亡的发生。结论 Hyp可以减轻大鼠心肌缺血再灌注损伤和心肌细胞凋亡。作用的机制可能与其抗氧自由基和NO自由基的形成，减少心肌缺血再灌注凋亡的发生有关。     

Protective effects of Acanthopanax Radix extract against endotoxemia induced by lipopolysaccharide

Endotoxemia causes an enhanced production of reactive oxygen radicals, which contribute to multiple organ dysfunction. When rats were given intravenous lipopolysaccharide and tested 6 h later we found that the activities of catalase and glutathione peroxidase (GSH-Px) in kidney, were acutely suppressed while in serum the levels of nitric oxide (NO), lipid peroxidation, urea nitrogen and creatinine were significantly increased, indicating the excessive production of reactive oxygen radicals and the presence of renal injury. Pretreatment of rats with Acanthopanax Radix extract administered orally for 30 days reduced the NO and lipid peroxidation levels, increased the activities of catalase and GSH-Px, and attenuated the renal dysfunction. These results suggested that scavenging of reactive oxygen radicals is part of the mechanism by which Acanthopanax Radix extract works as an effective agent in preventing multiple organ dysfunction.     

Protective effects of sarsasapogenin against early stage of diabetic nephropathy in rats

Rhizome of Anemarrhena asphodeloides Bunge (AA, family Liliaceae) has been widely used in China for thousands of years to treat febrile diseases and diabetes. Steroidal saponins from AA show good antidiabetes effects and ameliorate diabetic complications. This study was designed to investigate the effects of sarsasapogenin (Sar), a major sapogenin from AA, on diabetic nephropathy (DN) in rats, and to explore the possible mechanisms. Diabetic rats were divided into 3 groups treated orally with Sar (0, 20, or 60 mg/kg) and carboxymethylcellulose sodium, whereas normal rats for Sar (0 or 60 mg/kg) and carboxymethylcellulose sodium. We found that chronic treatment with Sar for 9 weeks significantly ameliorated renal dysfunction of diabetic rats, as evidenced by decreases in albuminuria, kidney weight index, serum uric acid, and morphologic changes such as extracellular matrix expansion and accumulation (fibronectin and collagen IV levels, etc.). Meanwhile, Sar treatment resulted in decreases in interleukin‐18, NLRP3, and activated caspase 1 levels as well as advanced glycation endproducts (AGEs) and their receptor (RAGE) levels in the renal cortex of diabetic rats. However, Sar has no effects on the above indices in the normal rats. Therefore, Sar can markedly ameliorate diabetic nephropathy in rats via inhibition of NLRP3 inflammasome activation and AGEs–RAGE interaction.     

Antioxidant activities of aqueous leaf extracts of Toona sinensis on free radical-induced endothelial cell damage

Ethnopharmacological relavence

In Taiwan, Toona sinensis (Toona sinensis) is well known as a traditional Chinese medicine, while the underlying pharmacological mechanisms of this drug are still a matter of debate.

Materials and methods

The purpose of this study was to evaluate the protective effects of non-cytotoxic concentrations of aqueous leaf extracts of Toona sinensis (TS extracts; 50-100 μg/mL) and gallic acid (5 μg/mL), a major component of these extracts, against AAPH-induced oxidative cell damage in human umbilical vein endothelial cells (ECs).

Results

Exposure of ECs to AAPH (15 mM) decreased cell viability from 100% to 43%. However, ECs were pre-incubated with TS extracts prior to AAPH induction resulted in increased resistance to oxidative stress and cell viability in a dose-dependent manner. An increase in ECs-derived PGI₂ and IL-1β in response to AAPH exposure was positively correlated with cytotoxicity and negatively with TS extracts concentrations. In addition, gallic acid also suppressed PGI₂ and IL-1β production in AAPH-induced ECs. Notably, TS extracts/gallic acid treatment significantly inhibited ROS generation, MDA formation, SOD/catalase activity, and Bax/Bcl-2 dysregulation in AAPH-stimulated ECs. Pretreatment of ECs with TS extracts/gallic acid also suppressed AAPH-induced cell surface expression and secretion of VCAM-1, ICAM-1 and E-selectin, which was associated with abridged adhesion of U937 leukocytes to ECs. Moreover, TS extracts/gallic acid treatment significantly inhibited the AAPH-mediated up regulation of PAI-1 and down regulation of t-PA in ECs, which may decrease fibrinolytic activity.

Conclusions

Therefore, Toona sinensis may possess antioxidant properties that protect endothelial cells from oxidative stress. Our results also support the traditional use of Toona sinensis in the treatment of free radical-related diseases and atherosclerosis.     

木犀草素对对乙酰氨基酚诱导的L02肝细胞损伤的保护作用

探讨木犀草素(luteolin,Lut)对对乙酰氨基酚(acetaminophen,APAP)诱导的L02肝细胞损伤的保护作用。CCK-8法检测Lut对L02细胞活性的影响;筛选APAP诱导L02细胞损伤的浓度及作用时间;细胞形态学、CCK-8实验和流式细胞术检测分析Lut对APAP诱导的L02细胞凋亡的影响;比色法检测细胞上清液中丙二醛(MDA)含量、谷胱甘肽(GSH)及超氧化物歧化酶(SOD)活性;RT-PCR检测凋亡相关基因Bax,Bcl-2,caspase-3的表达。结果显示Lut在2.5~40μmol·L~(-1)不影响L02细胞活性;12 mmol·L-1APAP作用于L02细胞12 h可用于建立肝细胞损伤模型。与模型组相比,Lut组细胞状态明显改善,胞体增大,贴壁能力恢复明显;细胞凋亡率明显下降;MDA含量显著下降(P0.05或P0.01),GSH和SOD活性显著提高(P0.05或P0.01),同时能够上调Bcl-2及下调Bax,caspase-3 mRNA的表达(P0.05或P0.01)。该实验证明了Lut对APAP诱导的L02细胞损伤有保护作用,其机制可能与其减轻氧化应激反应和抑制细胞凋亡有关。     

Protective effects of Garcinia kola seed extract against paracetamol-induced hepatotoxicity in rats

The hepatoprotective effect of Garcinia kola seed extract was investigated in rats treated with high doses of paracetamol. The extracts when administered at 100 mg/kg three times a day for five consecutive days reduced paracetamol- (800, 1000, 1200 mg/kg) induced lethality from 50, 90 and 100% to 0, 20 and 40%, respectively. There was a significant reduction in the liver enzymes SGOT and SGPT and histology scores. The hepatoprotective effect of the extract may be due to inhibition of cytochrome P-450 which normally converts paracetamol to the toxic intermediate metabolite N-acetyl-p-benzoquinoneimine (NAPQI).