Protective effect of Ligusticum chuanxiong and Angelica sinensis on endothelial cell damage induced by hydrogen peroxide

Ligusticum chuanxiong and Angelica sinensis have been widely used in traditional Chinese medicine to treat some pathological settings such as atherosclerosis and hypertension. We determined the protective effect of the extract of Ligusticum chuanxiong and Angelica sinensis (ELCAS) on human umbilical vein endothelial cells (ECV304) damage induced by hydrogen peroxide. ECV304 cells were pre-treated with ELCAS and exposed to 5 mM hydrogen peroxide. The results show that ELCAS dose- and time-dependently protected ECV304 cells against hydrogen peroxide damage and suppressed the production of reactive oxygen species (ROS). The decrement of ROS may be associated with increased activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX). Western blot analysis revealed that ELCAS significantly increased the phosphorylation of ERK and promoted eNOS expression. These observations indicate that ELCAS protected ECV304 cells against hydrogen peroxide damage by enhancing the antioxidative ability, activating ERK and eNOS signaling pathway. Our data also provide new evidence of Ligusticum chuanxiong and Angelica sinensis in preventing both cardiovascular and cerebrovascular diseases.     

当归对高脂血清所致ECV304细胞损伤的保护作用

实验观察了高脂血清对培养的人脐静脉内皮细胞（ECV304）的损伤及传统中药当归的保护作用,以探讨当归的抗动脉粥样硬化作用及其可能机制,培养人脐静脉内皮细胞,以高脂血清作损伤因子,用扫描电镜观察细胞的超微结构,分光光度法检测细胞培养液中一氧化氮（NO）的含量,免疫细胞化学方法检测细胞表面细胞间粘附分子－1（ICAM－1）,碱性成纤维细胞生长因子（bFGF)及转化生长因子β1（TGFβ）的表达,与高脂血清孵育24h后,内皮细胞的超微结构明显收损,且细胞表面ICAM－1,bFGF的表达明显增加,而细胞培养液中NO的量及细胞表达TGFβ1明显减少,加入当归后,高脂血清对内皮细胞的这些作用均可被逆转,当归对内皮细胞中ICAM－1,bFGF,TGFβ及NO表达改变的影响可能与其抗动脉粥样硬化的作用有关。     

当归对高脂血清所致ECV_(304)细胞损伤的保护作用

实验观察了高脂血清对培养的人脐静脉内皮细胞（ＥＣＶ３０４）的损伤及传统中药当归的保护作用,以探讨当归的抗动脉粥样硬化作用及其可能机制。培养人脐静脉内皮细胞,以高脂血清作损伤因子,用扫描电镜观察细胞的超微结构,分光光度法检测细胞培养液中一氧化氮（ＮＯ）的含量,免疫细胞化学方法检测细胞表面细胞间粘附分子－１（ＩＣＡＭ－１）、碱性成纤维细胞生长因子（ｂＦＧＦ）及转化生长因子β１（ＴＧＦβ１）的表达。与高脂血清孵育２４ｈ后,内皮细胞的超微结构明显收损,且细胞表面ＩＣＡＭ－１、ｂＦＧＦ的表达明显增加,而细胞培养液中ＮＯ的量及细胞表达ＴＧＦβ１明显减少。加入当归后,高脂血清对内皮细胞的这些作用均可被逆转。当归对内皮细胞中ＩＣＡＭ－１、ｂＦＧＦ、ＴＧＦβ１及ＮＯ表达改变的影响可能与其抗动脉粥样硬化的作用有关。     

Protective effect of ajoene on acetaminophen-induced hepatic injury in mice.

Ajoene, a garlic-derived sulfur-containing compound, exhibited a hepatoprotective effect against acetaminophen-induced liver injury in mice. A pretreatment with ajoene suppressed the rise in serum glutamic-pyruvic transaminase activity and the reduction in the hepatic reduced glutathione level. These effects of ajoene were observed dose-dependently (20-100 mg/kg). The pretreatment by ajoene also suppressed the decrease in hepatic protein thiol content resulting from acetaminophen administration.     

Protective effect of cornuside against carbon tetrachloride-induced acute hepatic injury

This study elucidated the effects of cornuside on carbon tetrachloride (CCl?)-induced hepatotoxicity. Rats were treated intraperitoneally with 0.5 mL/kg of CCl?. Sixteen h after CCl? treatment, the levels of serum aminotransferases, tumor necrosis factor-α (TNF-α), and lipid peroxidation were significantly elevated, whereas the hepatic antioxidative enzyme activities were decreased. These changes were attenuated by cornuside. Histological studies also indicated that cornuside inhibited CCl?-induced liver damage. Furthermore, the contents of hepatic nitrite, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) were elevated after CCl? treatment, while cytochrome P450 2E1 (CYP2E1) expression was suppressed. Cornuside treatment inhibited the formation of liver nitrite, and reduced the overexpression of iNOS and COX-2 proteins, but restored the liver CYP2E1 content as compared with the CCl?-treated rats. Our data indicate that cornuside protects the liver from CCl?-induced acute hepatotoxicity, perhaps due to its ability to restore the CYP2E1 function and suppress inflammatory responses, in combination with its capacity to reduce oxidative stress.     

Effect of polysaccharides from Angelica sinensis on gastric ulcer healing

Our previous study showed that a crude extract from Angelica sinensis (ASCE), which mainly consisted of polysaccharides, significantly promoted migration and proliferation of normal gastric epithelial cells. These results strongly suggest that ASCE has a direct wound healing effect on gastric mucosa. However, there is no report concerning the effect of ASCE on gastric ulcer healing in animal models. In this study, we found that ASCE promoted ulcer healing. The area of the ulcer was reduced. This was accompanied with a significant increase in mucus synthesis when compared with the control. Angiogenesis was inhibited by the treatment of ASCE. Cell proliferation, ODC and EGFR protein expression was not affected in this process. Thus, the mechanism of how ASCE accelerates ulcer healing in addition to its effect on mucus synthesis remains to be investigated.     

Immunomodulatory activity of polysaccharide isolated from Angelica sinensis

The immunomodulatory activities of an Angelica sinensis polysaccharide (AP), purified from the fresh root of A. sinensis Diels, were investigated in vitro in relation to the specificity to immune cells. AP consisted of rhamnose, arabinose, mannose, glucose, galactose with the molar ratio of 1.00:4.54:2.98:11.09:7.45. Cell proliferation results showed that proliferation of total spleen cells, macrophages and T cells were promoted by the action of AP. The treatment of AP increased the production of IL-2 and IFN-γ, while that of IL-4 was decreased. RT-PCR analysis displayed that the IL-2 and IFN-γ gene expression were enhanced but the IL-4 gene expression was decreased. Some differences in cytokines secretion pattern were also detected, the expression of IFN-γ was rapidly augmented while that of IL-2 responded later. The flow cytometry results showed that the percentage of CD4⁺T cell in total spleen cells was remarkably increased by AP, while that of CD8⁺T cell was slightly decreased. In conclusion, AP has immunomodulatory activity by regulating expression of Th1 and Th2 related cytokines. The time–effect relation of cytokines response also suggests that macrophages and natural killer cells involved in nonspecific immunity were primary activated, and helper T cell were secondarily affected by AP.     

银杏叶提取物对对乙酰氨基酚诱导的小鼠急性肝损伤的保护作用

目的：探究银杏叶提取物（GBE）对对乙酰氨基酚（APAP）诱导的小鼠急性肝损伤的保护作用及其机制。方法：30只小鼠随机分为对照组、模型组、GBE低、中、高剂量组（50,100,and 200 mg·kg^-1）,每组6只。除对照组外,剩余小鼠腹腔注射APAP （300 mg/kg）一次,随后GBE低、中、高剂量组按照相应剂量灌胃给药,治疗2 d后取材。观察各组肝脏大体情况和肝组织的病理组织学变化;取血测定各组小鼠血清中ALT、AST的活性和TNF-α、IL-6的水平;取肝检测各组肝组织中SOD、MPO的活性和GSH、MDA的含量;通过Western blot检测各组肝组织中Nrf2、HO-1蛋白的表达量。结果：与对照组相比,模型组肝脏明显肿大,病理表现差,血清中ALT、AST、TNF-α、IL-6的水平显著升高（P<0.01）,肝组织中GSH的含量和SOD的活性显著降低（P<0.01）,MDA的含量和MPO的活性显著升高（P<0.01）,Nrf2、HO-1蛋白表达明显下调（P<0.01）。与模型组相比,GBE组肝脏肿大减轻,病理表现有所改善,血清中ALT、AST、TNF-α、IL-6的水平显著降低（P<0.01）,肝组织中GSH的含量和SOD的活性显著提高（P<0.01）,MDA的含量和MPO的活性显著降低（P<0.01）,Nrf2、HO-1蛋白表达上调（P<0.05）,其中高剂量GBE组治疗效果最明显。结论：GBE可对APAP诱导的小鼠急性肝损伤具有保护作用,其作用机制可能是通过Nrf2/HO-1抗氧化途径发挥作用。     

Protective effect of berberine chloride on Plasmodium chabaudi-induced hepatic tissue injury in mice

The present study aimed to investigate the protective role of berberine (BER) against Plasmodium chabaudi-induced infection in mice. Animals were divided into three groups. Group I served as a vehicle control. Group II and group III were infected with 1000 P. chabaudi infected erythrocytes. Group III was gavaged with 100 μl of 10 mg/kg berberine chloride for 10 days. All mice were sacrificed at day 10 post-infection. The percentage of parasitemia was significantly reduced more than 30%, after treatment of mice with BER. Infection caused marked hepatic injuries as indicated by histopathological alterations as evidenced by the presence of hepatic lobular inflammatory cellular infiltrations, dilated sinusoids, vacuolated hepatocytes, increased number of Kupffer cells and the malaria pigment, hemozoin. These changes in livers led to the increased histological score. Also, infection induced a significant increase in liver alanine aminotransferase and aspartate aminotransferase and a significant increase in the total leucocytic count. Moreover, mice became anemic as proved by the significant decrease in erythrocyte number and haemoglobin content. BER showed a significant protective potential by improving the above mentioned parameters. Based on these results, it is concluded that berberine could offer protection against hepatic tissue damage.     

Protective effect of N-acetylcysteine in isoniazid induced hepatic injury in growing rats

Status of oxidative/antioxidative profile was the mechanistic approach to inumerate the nature of protection by N-acetylcysteine (NAC) in isoniazid (INH) exposed experimental animals. Analysis of lipid peroxidation, thiol levels, cytochrome P450, superoxide dismutase (SOD), catalase, glutathione peroxidase, reductase and transferase were estimated in liver along with the body and liver weight of animals and histological observations. Isoniazid exposure to animals resulted in no change in body and liver weights. Thiols, lipid peroxidation, catalase, SOD glutathione peroxidase, reductase, transferase and cytochrome P450 levels were altered with INH exposure. Supplementation of NAC with INH protected the animals against hepatotoxic reactions by minimizing the free radical induced tissue injury and overall maintenance of the endogenous scavengers of free radicals.     

Protective effect of aqueous garlic extract against lead-induced hepatic injury in rats

Effect of aqueous extract of garlic on hepatic injury due to lead-induced oxidative stress in experimental rats has been investigated. Lead acetate (LA) at a dose of 15 mg/kg body wt was administered ip to rats for 7 consecutive days to induce hepatic injury. Freshly prepared aqueous garlic extract (AGE) at a dose of 50 mg/kg body wt was fed orally to rats 1 h before LA treatment for similar period. LA treatment caused hepatic injury as evident from increased activities of serum glutamate pyruvate transaminase (SGPT) and alkaline phosphatase (ALP), increased serum bilirubin level and damage in the tissue morphology. Lead-induced oxidative stress in liver was evident from increased levels of lipid peroxidation and reduced glutathione. The decreased activity of superoxide dismutase (SOD) and an increased activity of catalase as well as an increased activity of xanthine oxidase (XO) indicate generation and possible accumulation of reactive oxygen intermediates. Furthermore, altered activities of lactate dehydrogenase (LDH), isocitrate dehydrogenase (ICDH), alpha-keto glutarate dehydrogenase (alpha-KGDH) and succinate dehydrogenase (SDH) also indicate an impaired substrate utilization and generation of oxidative stress. All these changes were found to be mitigated when the rats were pre-treated with the AGE. Results indicate that AGE has the potential to ameliorate lead-induced hepatic injury due to oxidative stress in rats. The protective effects may be due to the antioxidant properties of AGE and may have future therapeutic relevance.     

Protective effect of polysaccharide fractions from Radix A. sinensis against tert-butylhydroperoxide induced oxidative injury in murine peritoneal macrophages

Three Angelica sinensis polysaccharide fractions (APFs), named APF1, APF2 and APF3, were isolated and purified from Radix A. sinensis and their antioxidant activities were evaluated in isolated mouse peritoneal macrophages by pretreatment with APFs before exposure to 0.2 mM tertbutylhydroperoxide (t-BHP). The results showed that pretreatment of the macrophages with APFs as low as 10 microg/ml could significantly enhance t-BHP-decreased cell survival, intracellular glutathione (GSH) content and superoxide dismutase (SOD) activity, and also inhibited t-BHP-increased lactate dehydrogenase (LDH) leakage and malondialdehyde (MDA) formation (p < 0.05), and APF3 was the most active fraction, followed by APF2 and APF1 in decreasing order. Furthermore, we found for the first time that the bound-protein in APF3 was associated closely with the protective effects and the polysaccharide inhibited the excess NO release from t-BHP-activated macrophages to protect host cells.     

Protective effect of ozone treatment on the injury associated with hepatic ischemia-reperfusion: Antioxidant-prooxidant balance

The effects of ozone treatment on the injury associated to hepatic ischemia-reperfusion (I/R) was evaluated. Ozone treatment (1 mg/kg daily during 10 days by rectal insufflation) is shown to be protective as it attenuated the increases in transaminases (AST, ALT) and lactate levels observed after I/R. I/R leads to a decrease in endogenous antioxidant (SOD and glutathione) and an increase in reactive oxygen species (H₂O₂) with respect to the control group. However, ozone treatment results in a preservation (glutathione) or increase (SOD) in antioxidant defense and maintains H₂O₂ at levels comparable to those in the control group. The present study reports a protective effect of ozone treatment on the injury associated to hepatic I/R. The effectiveness of ozone could be related to its action on endogenous antioxidants and prooxidants balance in favour of antioxidants, thus attenuating oxidative stress.     

Protective effect of ozone treatment on the injury associated with hepatic ischemia-reperfusion: Antioxidant-prooxidant balance

The effects of ozone treatment on the injury associated to hepatic ischemia-reperfusion (I/R) was evaluated. Ozone treatment (1 mg/kg daily during 10 days by rectal insufflation) is shown to be protective as it attenuated the increases in transaminases (AST, ALT) and lactate levels observed after I/R. I/R leads to a decrease in endogenous antioxidant (SOD and glutathione) and an increase in reactive oxygen species (H₂O₂) with respect to the control group. However, ozone treatment results in a preservation (glutathione) or increase (SOD) in antioxidant defense and maintains H₂O₂ at levels comparable to those in the control group. The present study reports a protective effect of ozone treatment on the injury associated to hepatic I/R. The effectiveness of ozone could be related to its action on endogenous antioxidants and prooxidants balance in favour of antioxidants, thus attenuating oxidative stress.     

当归种子的生态学特性

当归种子在自然条件下,存放一年后,即失去发芽力,这一问题成为推广“立秋直播”新技术的严重障碍。为查明导致发芽力迅速下降的主要原因,探讨保持发芽力的有效措施,特对当归种子在自然条件下,发芽力的年变化,以及种子含水量(4—20%)、贮种温度(-10一33℃)和发芽温度(5—35℃)等对种子发芽力的影响进行了比较系统的试验分析。结果表明,当归种子属于“耐干藏种子类型”,高温和高湿是降低发芽力的主要因素。若将种子含水量降低到5—8%,并配合以较低的温度(15℃以下),发芽力则可长达三年以上。在当归产区,若能改“常温堆藏法”为“低温、干燥、密闭贮存法”,便可达到妥善保存当归种子的目的。     

阿的平保护纹状体神经元免受热处理损伤作用的研究

目的:探讨预先使用阿的平对热处理所致的纹状体神经元损伤的保护作用及其机制,为热环境致细胞损伤的干预和药物研究提供参考.方法:采用培养的大鼠纹状体神经元,培养液中加入不同浓度的阿的平,给予43℃热处理1h.台盼蓝染色鉴定细胞坏死,活性Caspase-3免疫染色和原位末端转移酶法评价细胞凋亡.结果:热处理明显影响神经元存活,导致细胞死亡,其中主要由细胞坏死过程介导;单独的阿的平对细胞毒性作用较小,预先使用阿的平能够减少热处理引起的细胞坏死.结论:阿的平对热环境致纹状体神经元的损伤有保护作用,这种保护作用主要通过减少细胞坏死过程介导.     

Protective effect of ulinastatin on hepatic ischemia reperfusion injury through autophagy activation in Chang liver cells

This study aimed to investigate the protective effect of ulinastatin in hepatic ischemia-reperfusion progress, involving its association with the role of autophagy during hypoxia-induced hypoxia-reoxygenation injury in vitro. The model of hepatic hypoxia/reoxygenation (H/R) injury in Chang liver cells was established. After treatment with ulinastatin at the doses of 10, 100, and 1000 U/mL in H/R liver cells, the cell proliferation was significantly increased, morphological damage was reduced, and the cell apoptosis rate was decreased. The protein levels of antiapoptotic myeloid cell leukemia-1 (Mcl-1) and caspase-3 were upregulated, and C-PARP protein was downregulated. Meanwhile, ulinastatin led to an increase in the messenger RNA and protein levels of autophagy maker Unc-like kinase 1 (ULK1), Beclin-1, and microtubule-associated protein 1 light chain 3 (LC-3) and a decrease in p62. Then, 3-methyladenine (3-MA), an inhibitor of autophagy, made morphological damage and cell apoptosis worsen in ulinastatin-treated H/R liver cells. And the expression levels of caspase-3, C-PARP, p62, Beclin-1, and LC-3, proteins were also reversed by 3-MA. Taken together, our results demonstrate that ulinastatin inhibited the hepatic H/R injury in Chang liver cells, which was, to some extent, related to the autophagy activation.     

GABAergic phthalide dimers from Angelica sinensis (Oliv.) Diels

The methanol extract of Angelica sinensis (Oliv.) Diels roots (Dang Gui) has been shown to exhibit competitive binding to the GABAa receptor, suggesting the presence of GABAergic ligands. Chromatographic fractionation of the methanol extract led to the isolation of two GABAergic dimeric phthalides 1 and 2. Gelispirolide (1) was elucidated as a new phthalide dimer composed of a Z-ligustilide and a Z-butylidenephthalide unit on the basis of spectroscopic approaches including one- and two-dimensional NMR, HRESIMS and HRESIMS-MS. Compound 2 was identified as the known dimeric phthalide, riligustilide, by comparison of its spectroscopic data with literature values. Its dimeric linkage and stereochemistry were ascertained by a single crystal X-ray diffraction experiment. Both dimers 1 and 2 were found to be active in an in vitro GABAa receptor-binding assay with IC50 values of 29 and 24 microM, respectively.     

Antivirus and immune enhancement activities of sulfated polysaccharide from Angelica sinensis

This study is to synthesize sulfated Angelica polysaccharides (APSs) and investigate the activity of one of the sulfated derivatives APS-1 on murine leukemia virus in vivo. Six sulfated derivatives with degree of sulfation ranging from 0.68 to 1.91 were obtained. And the virus replication was inhibited by APS-1 at the dose of 10 and 30 mg/kg (26% and 30% inhibition respectively). Furthermore, both the percentage of CD4(+) cells and CD4(+)/CD8(+) ratio in peripheral blood cells were significantly enhanced by APS-1 at 3-30 mg/kg. In addition, the reduced thymus/body weight index by murine leukemia virus infection was increased by ASP-1 in a dose dependent manner. These results suggest that APS-1 could not only inhibit virus replication, but also improve the immune function. APS-1 may be a potential new and better antiviral drug.