三七总皂苷对细胞免疫炎症因子作用的研究进展

三七总皂苷（PNS）系从我国中药材三七Panaxnotoginseng（Burk）F．H．Chen的根茎中提取出来的主要药理成分,主要由人参皂苷Rg1、Rb1,三七皂苷R1、R2等40多种皂苷组成。三七总皂苷是中药三七的主要活性成分。     

短葶山麦冬多糖和皂苷成分的研究进展

对近年来研究短葶山麦冬的文献进行了回顾分析,对其主要的多糖、皂苷类成分的化学结构、药理活性、提取分离和含量测定等进行了总结,发现短葶山麦冬有广泛的研究价值和应用前景。     

短葶山麦冬总多糖对小鼠免疫功能的影响

目的观察短葶山麦冬总多糖对小鼠免疫功能的影响。方法短葶山麦冬多糖以0.11、0.33、1.0g·kg-1 3个剂量对小鼠连续灌胃30d,检测短葶山麦冬总多糖对小鼠碳廓清能力、巨噬细胞吞噬鸡红细胞能力及NK细胞活性等免疫指标。结果短葶山麦冬总多糖对胸腺指数、碳廓清能力、巨噬细胞吞噬鸡红细胞能力、NK细胞活性均有显著作用。结论短葶山麦冬总多糖能增强小鼠非特异性免疫功能。     

基于药物相互作用研究麦冬皂苷D对麦冬皂苷D′急性和亚急性毒性的影响

目的 基于药物相互作用研究麦冬皂苷D(OPD)对麦冬皂苷D′(OPD′)急性和亚急性毒性的影响。方法 (1)采用Bliss法测定急性毒性：小鼠分为OPD′单用、OPD+OPD′(OPD和OPD′提前混合)和OPD→OPD′(先注射OPD间隔15 min再注射OPD′)组,其中OPD′单用组5个剂量分别为3.127,6.460,9.979,10.245和13.098 mg·kg^-1;OPD+OPD′组OPD和OPD′1∶1混合,5个剂量分别为5.711,6.129,7.626,8.712和10.262 mg·kg^-1;OPD→OPD′组OPD固定为1.0 mg·kg^-1,OPD′5个剂量分别为11.351,15.551,22.732,26.568和30.137 mg·kg^-1。单次尾静脉注射,观察给药后14 d内小鼠的行为表征和死亡数。(2)亚急性毒性试验：SD大鼠随机分为5组,分别为正常对照、OPD单用(0.25 mg·kg^-1)、OPD′单用(0.25 mg·kg^-...     

三七总皂苷对血小板衍生生长因子诱导的血管平滑肌细胞增殖的影响

目的 研究三七总皂苷（TPNS）对血小板衍生生长因子（PDGF）诱导SD大鼠胸主动脉平滑肌细胞（tASMC）增殖的抑制作用及机制.方法 将SD大鼠tASMC进行体外培养,采用四甲基偶氮唑盐比色法检测TPNS对PDGF诱导平滑肌细胞增殖的影响,采用钙荧光指示剂Fura-2/AM负载的平滑肌细胞测定细胞内游离钙浓度.结果 PDGF能诱导血管平滑肌细胞增殖（P〈0.01）,并使细胞内游离钙浓度增高（P〈0.01）,TPNS能抑制该作用（P〈0.01）.结论 TPNS可能通过抑制细胞内游离钙离子浓度的升高来拮抗PDGF诱导的平滑肌细胞增殖.     

三七总皂苷对血小板衍生生长因子诱导的血管平滑肌细胞增殖的影响

目的 研究三七总皂苷(TPNS)对血小板衍生生长因子(PDGF)诱导SD大鼠胸主动脉平滑肌细胞(tASMC)增殖的抑制作用及机制.方法 将SD大鼠tASMC进行体外培养,采用四甲基偶氮唑盐比色法检测TPNS对PDGF诱导平滑肌细胞增殖的影响,采用钙荧光指示剂Fura-2/AM负载的平滑肌细胞测定细胞内游离钙浓度.结果 PDGF能诱导血管平滑肌细胞增殖(P<0.01),并使细胞内游离钙浓度增高(P<0.01),TPNS能抑制该作用(P<0.01).结论 TPNS可能通过抑制细胞内游离钙离子浓度的升高来拮抗PDGF诱导的平滑肌细胞增殖.     

三七总皂苷对内毒素损伤血管内皮细胞炎症特性的影响

目的:研究三七总皂苷对内毒素损伤血管内皮细胞炎症特性的影响.方法:随机将培养人脐静脉内皮细胞(ECV304)分为对照组、内毒素组、用药组(内毒素加三七总皂苷)组,各组细胞间粘附分子-1表达用流式细胞仪测定、培养上清中血小板活化因子用生物法测定、培养上清中一氧化氮的变化用Griess法测定.结果:内毒素加三七总皂苷组与内毒素组相比,各指标均较内毒素组有不同程度降低,但仍高于对照组.结论:三七总皂苷对内皮细胞炎症介质产生及粘附分子表达有抑制作用,提示其具有抗炎作用.     

不同材质及孔径超滤膜对麦冬皂苷类成分的影响

目的 考查不同材质及不同孔径的超滤膜对5种麦冬皂苷类成分的影响。方法 以麦冬皂苷C、麦冬皂苷D、麦冬皂苷D''、麦冬皂苷Ra、龙血苷F为考察指标,利用HPLC-UV法测定其超滤前后变化,考察不同材质及孔径的超滤膜对5种麦冬皂苷收率的影响以及各超滤阶段对麦冬皂苷类成分截留率的变化情况。结果 不同材质及不同孔径的超滤膜处理下,麦冬皂苷类成分的收率不同;聚醚砜材质超滤膜对各超滤阶段的麦冬皂苷类成分截留率变化趋势与纤维素材质是一致的。结论 采用100 kDa的聚醚砜超滤膜,可以使麦冬皂苷类成分的收率达到最高,用50%乙醇进行等体积透析,会对麦冬皂苷的收率有较大的改善。     

麦冬多糖对正常和实验性糖尿病小鼠血糖的影响

目的：研究麦冬多糖对正常和实验性糖尿病小鼠血糖的影响。方法：昆明种小鼠４０只，分成４组，分别用麦冬多糖（１００和３００ｍｇ／ｋｇ）、优降糖（２．５ｍｇ／ｋｇ）和等体积生理盐水（２ｍｌ／只）灌胃，测定各组正常小鼠的血糖水平及以葡萄糖（２ｇ／ｋｇ）、四氧嘧啶（７０ｍｇ／ｋｇ）、肾上腺素（０．０２ｍｇ／ｋｇ）所致小鼠高血糖模型的血糖水平。结果：剂量为１００和３００ｍｇ／ｋｇ的麦冬多糖灌胃对葡萄糖、四嘧啶     

人参总皂苷在巨噬细胞RAW264.7中降低脂多糖诱导的炎症和氧化应激

目的:通过脂多糖诱导巨噬细胞RAW264.7建立炎症和氧化应激模型,探讨人参总皂苷的抗炎和抗氧化效果以及对脂多糖诱导的巨噬细胞RAW264.7自噬水平的影响。方法:培养RAW264.7细胞,给予脂多糖刺激并加入不同浓度人参总皂苷,使用硝酸还原酶法检测细胞内一氧化氮水平;ELISA法检测小鼠肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)的分泌;DCFH-DA荧光探针法检测人参总皂苷处理后活性氧的变化;超氧化物阴离子荧光探针(dihydroethidium, DHE)检测细胞内超氧化物阴离子水平;吖啶橙染色法检测自噬体的形成;Western Blot法检测COX-2、NF-кB、Nrf2、HO-1、GCLC、GCLM、Beclin1、ATG7和LC3的蛋白表达。结果:与脂多糖模型组相比,随着TGS的浓度增加,细胞一氧化氮水平、TNF-α的分泌降低,细胞内活性氧和超氧阴离子的含量降低,巨噬细胞RAW264.7酸性自噬体的数量增加;Nrf2、HO-1、GCLC、GCLM、Beclin1、ATG7及LC3的蛋白表达提高,COX-2、NF-кB的蛋白表达降低。结论:在...     

香烟提取物对人支气管上皮细胞抗衰老因子Klotho的影响分析及应对措施的效果研究

目的分析香烟提取物对人支气管上皮细胞(HBE)抗衰老因子 Klotho的影响及应对措施的效果。方法选择 2016年 3月至 2017年 8月在仪陇县人民医院进行体检的健康人员 30例作为对照组,选择同期在该院治疗的具有 10年吸烟史慢性阻塞性肺疾病(简称“慢阻肺”)病人 30例作为观察组,对两组人群外周血细胞因子白细胞介素(IL)?8、IL?6、肿瘤坏死因子(TNF)?α、 Klotho蛋白表达水平进行评估。同时以香烟提取物刺激 HBE细胞构建支气管炎症模型,并采用地塞米松进行干预。结果慢阻肺病人外周血 IL?8,IL?6及 TNF?α含量均显著高于对照组,差异有统计学意义(t＝3.012,P＝0.013;t＝4.092,P＝0.000;t＝     

芹菜素对烟草烟雾提取物诱导血管内皮细胞损伤的作用

目的探讨芹菜素改善烟草烟雾提取物诱导的血管内皮细胞损伤的机制。方法培养人脐静脉内皮细胞株,给予烟雾提取物(CSE)模拟吸烟环境和不同浓度芹菜素。实验分为正常对照组、CSE组、芹菜素组,检测细胞存活率、凋亡率、细胞内活性氧ROS水平,以及细胞上清液中IL-6、TGF-β、INF-γ各因子的表达。结果芹菜素在24 h和48 h均能显著缓解不同浓度CSE诱导的内皮细胞存活率的降低,降低不同浓度CSE诱导的内皮细胞的凋亡率,同时芹菜素可抑制CSE诱导的细胞内ROS水平,以及上清液中炎性因子IL-6、TGF-β、INF-γ的升高,差异均有统计学意义(P<0.05,P<0.01)。结论芹菜素以血管内皮细胞凋亡和炎性反应为标靶,抑制吸烟引起的血管内皮细胞损伤,为临床治疗提供理论依据。     

羧甲司坦对香烟烟雾提取物诱导A549细胞炎性损伤的影响

目的：探讨羧甲司坦抑制香烟烟雾提取物（CSE）诱导A549细胞炎性损伤的作用。方法：传代培养人A549细胞并分为5组：对照组,CSE组,羧甲司坦低、中、高剂量组（予不同浓度羧甲司坦孵育和CSE诱导）。qRT-PCR、ELISA检测主要细胞因子的合成和释放,免疫蛋白印迹（Western-blot）、免疫荧光（IF）观察NF-κB及MAPK相关信号通路的活化情况。结果：以羧甲司坦预处理或后处理,均可降低CSE诱导的A549细胞的IL-6、IL-8和MIP-1β mRNA的表达;降低IL-6及IL-8的释放。羧甲司坦预处理可抑制p65入核。Western-blot结果显示,对照组,CSE组,羧甲司坦低、中、高剂量组的P-p65蛋白相对表达量分别为（0.17±0.05）、（0.90±0.19）、（0.68±0.15）、（0.64±0.12）和（0.57±0.13）,pERK1/2蛋白相对表达量分别为（0.30±0.10）、（1.25±0.33）、（1.01±0.19）、（0.89±0.22）和（0.81±0.18）,CSE组均明显高于对照组,羧甲司坦低、中、高剂量组均明显低于CSE组（P<0.05）。结论：羧甲司坦通过抑制NF-κB p65及ERK1/2 MAPK活化,发挥对CSE诱导的A549细胞炎性损伤的保护作用。     

Effect of cigarette smoke extract on neonatal porcine vascular smooth muscle cells

Rationale: Cigarette smoke exposure in the perinatal period increases the risk of various prenatal and postnatal complications, including sudden infant death syndrome (SIDS) and persistent pulmonary hypertension of the newborn (PPHN). We investigated the cellular effects of cigarette smoke extract (CSE) in the developing vasculature. Methods: Vascular smooth muscle cells (VSMC) were isolated from neonatal porcine carotid arteries, splenic arteries, and main and resistance pulmonary arteries. Effects of CSE on VSMC proliferation, viability, apoptosis, and media nitrates and nitrites were evaluated. The effects of known constituents of CSE (nicotine, benzopyrene, acrolein, acetaldehyde), aged CSE, CSE with added hemoglobin, devolatilized CSE, CSE with added dithiothreitol (DTT), and reduced glutathione (GSH) on cell proliferation and viability were assessed. Results: CSE caused a dose- and time-dependent decrease in neonatal VSMC numbers isolated from all four sites, mainly as a result of increased cell necrosis and not apoptosis. Nitrates and nitrites were below the threshold of detection. Nicotine and benzopyrene did not affect cell counts, while acrolein and acetaldehyde decreased cell counts in a dose-dependent manner. Addition of hemoglobin, devolatilization, and the addition of DTT or GSH slightly decreased CSE inhibition. Conclusions: CSE causes necrosis of neonatal VSMC, and this toxicity is mediated mainly by volatile components such as acrolein and acetaldehyde, possibly in association with nitric oxide and carbon monoxide.     

槲皮素对香烟烟雾提取物诱导的人支气管上皮细胞死亡的抑制效应

目的研究槲皮素对香烟烟雾提取物(cigarette smoke extract,CSE)诱导的人支气管上皮细胞死亡的影响,并初步探讨其机制。方法槲皮素预处理人支气管上皮细胞(Be-as-2b)16h,然后以CSE刺激不同时间,倒置显微镜下观察细胞形态,以四甲基偶氮唑盐比色法(MTT)及Western blot分别检测细胞活力和血红素加氧酶-1(HO-1)蛋白的表达。结果槲皮素明显减少了CSE诱导的Beas-2b细胞死亡,并对CSE处理后细胞活力的降低具有明显的抑制作用;槲皮素还明显提高了CSE诱导的Beas-2b细胞中HO-1的蛋白表达水平,且呈浓度依赖性。结论槲皮素对于CSE诱导的人支气管上皮细胞死亡具有明显的抑制作用,其机制可能是通过提高细胞中CSE诱导的HO-1蛋白表达水平。     

Increased interleukin-8 expression by cigarette smoke extract in endothelial cells

It has been reported that cigarette smoking worsens alcohol-induced gastric lesions through neutrophil infiltration. We hypothesize that IL-8, a potent chemotactic factor for neutrophil is likely to be involved in this ulcerogenic process. To evaluate this phenomenon, the ability of cigarette smoke extract (CSE) to induce endothelial cell expression of IL-8 was examined. Two different fractions (ethanol or chloroform soluble extracts) of CSE with their chemical types identified showed a time- and dose-dependent increase on IL-8 secretion from ECV304 cell line. Protein kinase C (PKC) inhibitor GF109203X had no effect on IL-8 response in basal secretion and also to these stimuli. Protein tyrosine kinase (PTK) inhibitor genistein and protein kinase A (PKA) inhibitor H8 at respective concentrations significantly reduced chloroform and ethanol soluble extract-induced IL-8 expression by about 34 and 35% respectively at 8 h after incubation. It is concluded that CSE increases IL-8 release from human endothelial cells through PTK and PKA activation.     

Combination effects of cigarette smoke extract and ambient ultrafine particles on endothelial cells

Previous studies have shown that ambient ultrafine particles with diameters less than 100nm (UFPs) can pass from the lungs to the circulation because of their very small diameter, and induce lung oxidative stress with a resultant dysfunction of lung endothelial cells. However, no studies have addressed the potential combined effects of UFPs and cigarette smoke on vascular endothelial cells. We hypothesized that co-exposure to UFPs and cigarette smoke extract (CSE) may cause combined effects on activation of endothelial cells and dysfunction of endothelium by oxidative stress through activation of NADPH oxidase. We determined the effects of UFPs with or without CSE on mouse pulmonary microvascular endothelial cells (MPMVEC) obtained from C57BL/6J (wild-type) and gp91(phox) knock-out mice (gp91(phox) is one of the key components of NADPH oxidase, one of ROS generators). Our results showed that exposure of MPMVEC from wild-type mice to UFPs or CSE, at a non-toxic dose, induced reactive oxygen species (ROS) generation, increased phosphorylation of p38 and Erk1/2, and up-regulated early growth response -1 (Egr-1) and IL-6 genes. These effects were significantly enhanced when cells were co-exposed to both UFPs and CSE. However, exposure of MPMVEC from gp91(phox) knock-out mice did not induce the above effects. Furthermore, UFPs- and/or CSE-induced Egr-1 mRNA upregulation was attenuated significantly when cells were pre-treated with p38 specific inhibitor, SB 203580, or MEK1/2 inhibitor, PD98059, and Egr-1 siRNA treatment abolished UFPs- and/or CSE-induced overexpression of IL-6. Our results suggest that UFPs and/or CSE caused activation of NADPH oxidase, resulting in ROS generation that led to activation of MAPKs through induced phosphorylation of p38 and ERK1/2 MAPKs and upregulation of Egr-1. Those effects may further result in endothelial dysfunction through production of cytokines such as IL-6. Our results suggest that co-exposure to UFPs and CSE causes enhanced injury to endothelial cells.     

Hydrogen sulfide alleviates apoptosis and autophagy induced by beryllium sulfate in 16HBE cells

Beryllium and its compounds are systemic toxicants that are widely applied in many industries. Hydrogen sulfide has been found to protect cells. The present study aimed to determine the protective mechanisms involved in hydrogen sulfide treatment of 16HBE cells following beryllium sulfate-induced injury. 16HBE cells were treated with beryllium sulfate doses ranging between 0 and 300 μM BeSO₄. Additionally, 16HBE cells were subjected to pretreatment with either a 300 μM dose of sodium hydrosulfide (a hydrogen sulfide donor) or 10 mM DL-propargylglycine (a cystathionine-γ-lyase inhibitor) for 6 hr before then being treated with 150 μM beryllium sulfate for 48 hr. This study illustrates that beryllium sulfate induces a reduction in cell viability, increases lactate dehydrogenase (LDH) release, and increases cellular apoptosis and autophagy in 16HBE cells. Interestingly, pretreating 16HBE cells with sodium hydrosulfide significantly reduced the beryllium sulfate-induced apoptosis and autophagy. Moreover, it increased the mitochondrial membrane potential and alleviated the G2/M-phase cell cycle arrest. However, pretreatment with 10 mM DL-propargylglycine promoted the opposite effects. PI3K/Akt/mTOR and Nrf2/ARE signaling pathways are also activated following pretreatment with sodium hydrosulfide. These results indicate the protection provided by hydrogen sulfide in 16HBE cells against beryllium sulfate-induced injury is associated with the inhibition of apoptosis and autophagy through the activation of the PI3K/Akt/mTOR and Nrf2/ARE signaling pathways. Therefore, hydrogen sulfide has the potential to be a promising candidate in the treatment against beryllium disease.     

Telomere shortening and DNA damage of embryonic stem cells induced by cigarette smoke

Embryonic stem cells (ESCs) provide a valuable in vitro model for testing toxicity of chemicals and environmental contaminants including cigarette smoke. Mouse ESCs were acutely or chronically exposed to smoke components, cigarette smoke condensate (CSC), or cadmium, an abundant component of CSC, and then evaluated for their self-renewal, apoptosis, DNA damage and telomere function. Acute exposure of ESCs to high dose of CSC or cadmium increased DNA damage and apoptosis. Yet, ESCs exhibited a remarkable capacity to recover following absence of exposure. Chronic exposure of ESCs to low dose of CSC or cadmium resulted in shorter telomeres and DNA damage. Together, acute exposure of ESCs to CSC or cadmium causes immediate cell death and reduces pluripotency, while chronic exposure of ESCs to CSC or cadmium leads to DNA damage and telomere shortening. Notably, a sub-proportion of ESCs during passages is selected to resist to smoke-induced oxidative damage to telomeres.