葛根素对糖尿病大鼠白内障过程中晶状体诱导型一氧化氮合酶的影响

目的: 观察大鼠糖尿病性白内障(DC)晶状体诱导型一氧化氮合酶(iNOS)基因表达变化及葛根素对DC的治疗作用。方法: 经腹腔注射链脲佐菌素(STZ)复制大鼠DC模型。实验分为STZ组(STZ,45 mg/kg bw,ip)、葛根素组(STZ+葛根素,140 mg/kg bw ip)和对照组(等量生理盐水,ip),分别于实验第20、40、60 d摘取大鼠晶状体,用逆转录聚合酶链反应(RT-PCR)、蛋白印迹(Western blot)和生化方法检测晶状体iNOS mRNA和蛋白表达及一氧化氮(NO)含量、一氧化氮合酶(NOS)活性变化,观察晶状体损伤的宏观和微观病理变化。 结果:对照组大鼠晶状体透明,iNOS mRNA未见明显表达,蛋白呈微弱表达,NOS活性较低,NO含量较少。在DC形成过程中,晶状体出现混浊、晶状体上皮细胞(LEC)有明显病变,并随病程延长而加重;晶状体iNOS mRNA和蛋白表达明显上调,NOS活性增强、NO生成增加,并呈现时间依赖性。用药40-60 d时葛根素组前述晶状体病理变化轻于STZ组,iNOS mRNA和蛋白表达明显低于STZ组,NOS活性及NO 生成低于STZ组。结论: 在大鼠DC形成过程中晶状体iNOS 基因表达上调、NO含量增加;葛根素可减轻晶状体损伤,机制可能与抑制iNOS 基因表达、减少NO生成有关。     

葛根素对高血压病病人气管插管期心血管反应的影响

目的:探讨葛根素注射液控制高血压病病人插管期心血管反应的有效性.方法:30例原发性高血压全麻病人,随机均分为葛根素组(P组)和对照组(C组),分别于诱导前1 h静脉输注中药葛根素注射液500 mg或不用降压药,观察插管期收缩压(SBP)、舒张压(DBP)、心率(HR)、平均动脉压(MAP)和心率收缩压乘积(RPP).结果:与诱导前比较,两组诱导后SBP、DBP、MAP均明显下降(P＜0.01);C组插管时及插管后等时段SBP、DBP、MAP、HR及RPP均明显增加(P＜0.01或P＜0.05);P组插管时及插管后1 min SBP、DBP、MAP均有不同程度下降(P＜0.01或P＜0.05),HR及RPP未见明显变化.与C组比较,P组插管时及插管后1 min、3 min等时段SBP、HR及RPP均显著降低(P＜0.01或P＜0.05).结论:中药葛根素注射液静脉输注能有效控制插管刺激引起的心血管反应,有助于高血压病病人插管期维持血流动力学相对稳定.     

葛根素对两肾一夹高血压大鼠左室肥厚的影响

目的探讨葛根素对两肾一夹高血压大鼠左室肥厚的影响。方法清洁级雄性SD大鼠30只,随机均分为正常组、模型组和葛根素组,后两组大鼠采用两肾一夹方法(2K1C)制成肾性高血压动物模型。测定各组平均颈动脉压(mCAP)、左室舒张末期压(LVEDP)以及左心室压力变化最大速率(LV±dp/dtmax),测定左心重与体重比值(LVM/BW)。电子显微镜观测心肌形态学的变化。结果①与正常组相比,模型组大鼠mCAP、LVEDP以及LVM/BW分别升高36.58%(P<0.05)、333.8%(P<0.01)和20.24%(P<0.01),而LV+dp/dtmax则降低85.35%(P<0.05)。两组大鼠LV-dp/dtmax差异无统计学意义。②与模型组相比,葛根素组大鼠LVEDP、LVM/BW以及LV-dp/dtmax分别降低65.24%(P<0.05)、13.12%(P<0.05)和96.28%(P<0.01),LV+dp/dtmax则升高179.18%(P<0.01)。两组大鼠mCAP差异无统计学意义。③电镜观察:模型组大鼠心室肌丝松散,部分溶解断裂,闰盘增宽;基膜下轻度水肿;细胞核固缩现象易见。正常组和葛根素组未见。结论葛根素可以逆转两肾一夹高血压大鼠左室肥厚。     

葛根素与地高辛联合治疗对充血性心力衰竭大鼠的影响

目的探讨葛根素(Pur)与地高辛(DG)合用对充血性心力衰竭(CHF)大鼠血流动力学和心钠素(ANP)及血管紧张素Ⅱ(AngⅡ)的影响。方法采用腹主动脉缩窄的方法建立压力超负荷大鼠CHF模型,通过十六导生理记录仪测量心功能及血流动力学指标;采用放免法测定血浆ANP和AngⅡ的浓度。结果(1)模型组与假手术组相比心功能及血流动力学明显异常(P<0.01);(2)Pur与DG合用组能显著改善CHF大鼠的心功能及血流动力学异常;(3)Pur与DG合用能明显降低鼠CHF时ANP及AngⅡ的浓度(P<0.01)。结论Pur与DG合用能明显改善CHF大鼠的血流动力学及神经内分泌异常。     

葛根素对于快速心房颤动家兔模型心房颤动的治疗作用

目的 探讨葛根素对家兔快速心房颤动(AF)模型的干预效果.方法 24只白兔随机均分为快速心房起搏AF(A)组、快速心房起搏AF±葛根素40 mg/kg治疗(B)组和正常对照组(C)组.A组与B组连续刺激7d,AF稳定后开胸取左心房组织,检测房颤前后心房肌L-型钙通道α1、β1亚单位和人类电压门控性钾通道Kv4.3 mRNA的表达,用透射电子显微镜观察各组心房肌超微结构改变.结果 与C组比较,A、B组心房肌细胞L型钙通道α1、β1亚单位和钾通道Kv4.3 mRNA的表达水平均下调(P＜0.01).A组心房肌细胞肌纤维溶解,线粒体明显肿胀、变形,出现空泡,嵴排列紊乱、融合或消失;B组上述改变较A组减轻.结论 葛根素可以改善快速AF所致的心房电重构,对AF有一定的治疗效果.     

Puerarin ameliorates oxidative stress in a rodent model of traumatic brain injury

Background

A wealth of evidence has suggested that oxidative stress is involved in the secondary brain injury after traumatic brain injury (TBI). Recently, numerous in vivo and in vitro studies were reported that puerarin could inhibit oxidative stress through the activation of phosphatidylinositol 3-kinase (PI3K)-Akt pathway. It is unknown, however, whether puerarin can provide neuroprotection and reduce oxidative stress after TBI. The present study investigated the effects of puerarin on the TBI-induced neurodegeneration, oxidative stress, and the possible role of PI3K-Akt pathway in the neuroprotection of puerarin, in a rat model of TBI.

Materials and methods

Rats were randomly distributed into various subgroups undergoing the sham surgery or TBI procedures. Puerarin (200 mg/kg) was given intraperitoneally at 10 min before injury and PI3K-Akt pathway inhibitor LY294002 was also administered intracerebroventricular in one subgroup. All rats were killed at 24 h after TBI for examination.

Results

Our data indicated that puerarin could significantly reduce TBI-induced neuronal degeneration, accompanied by the partial restoration of the redox disturbance and enhanced expression of phospho-Akt in the pericontusional cortex after TBI. Moreover, PI3K-Akt pathway inhibitor LY294002 could partially abrogate the neuroprotection of puerarin in rats with TBI.

Conclusions

These results indicate that puerarin can ameliorate oxidative neurodegeneration after TBI, at least in part, through the activation of PI3K-Akt pathway.     

Puerarin protects pulmonary arteries from hypoxic injury through the BMPRII and PPARγ signaling pathways in endothelial cells

BackgroundRecent evidence indicates that Puerarin has a protective effect on pulmonary arteries. In the present study, we aimed to investigate whether Puerarin could protect pulmonary arterial endothelial cells from hypoxic injury and determine its potential targets.MethodsIn our study, human pulmonary arterial endothelial cells (HPAECs) were injured by hypoxic (1% O₂) incubation. Cell viability was detected by a cell counting kit (CCK8). The production of nitric oxide (NO) was detected by Griess reagent and endothelin-1 (ET-1) was detected by the ELISA method. Oxidative stress was measured by a fluorescence microscope via the fluorescent probe DCFH-DA. Western blotting was employed for studying the mechanism.ResultsThe results show that Puerarin protects HPAECs from hypoxia-induced apoptosis and slightly improves cell viability. Puerarin increases NO and decreases ET-1 to prevent the imbalance between vasoactive substances induced by hypoxia in HPAECs. Puerarin also inhibits the oxidative stress induced by hypoxia. The results from the Western blot show that Puerarin activates the BMPRII/Smad and PPARγ/PI3K/Akt signaling pathways.ConclusionIn conclusion, Puerarin protects HPAECs from hypoxic injury through the inhibition of oxidative stress and the activation of the BMPRII and PPARγ signaling pathways. This work provides insight into the development of Puerarin as a treatment for hypoxic pulmonary hypertension.     

葛根素对人肝癌细胞HepG2侵袭与迁移能力的影响

目的探索葛根素对人肝癌细胞HepG2侵袭与迁移能力的影响及其相关分子机制。方法将人肝癌细胞HepG2分为两组,实验组用60μg/ml、30μg/ml的葛根素进行培养,对照组用加入等体积的葛根素溶解剂二甲基亚砜(DMSO)的培养基培养;利用细胞划痕实验检测两组细胞的迁移能力;利用Transwell小室检测两组细胞的侵袭能力;通过蛋白免疫印迹检测波形蛋白(vimentin)、E-钙黏蛋白(E-cadherin)、基质金属蛋白酶2/9(matrix metalloproteinase2/9,MMP2/9)。结果人肝癌细胞HepG2在经过葛根素培养后,细胞划痕的愈合能力下降,Transwell小室细胞的侵袭能力下降;E-cadherin表达上升(P<0.05),vimentin、MMP2和MMP9的表达下调(P<0.05)。结论葛根素通过上皮细胞间质转化(EMT)途径导致人肝癌细胞HepG2侵袭转移能力下降。     

葛根中葛根素的树脂纯化工艺研究

目的：探索葛根中有效成分葛根素的纯化方法,制备葛根提取物,提取物中葛根素含量应不得少于40％。方法通过对比试验确定树脂型号、用量及上样流速,用正交试验法确定树脂洗脱的条件。结果葛根经提取浓缩过滤后,上D-101大孔树脂吸附,1 g葛根药材（葛根素含量为2.71%）的提取液需用2.7 g D-101大孔树脂吸附,选择12 mL/min流速作为上样速度,选择20 mL/min水洗10倍柱体积的水清洗,以25%乙醇洗脱,收集洗脱液,浓缩至稠膏,减压干燥,所得葛根提取物葛根素含量均在40%以上。结论该法提取葛根中有效成分葛根素较稳定、可靠,适合工业生产。     

抗眩颗粒的制备及葛根素的含量测定

目的：探讨抗眩颗粒的制备和葛根素的含量测定方法.方法：采用水提浓缩制粒制备抗眩颗粒,采用高效液相色谱法对方中葛根的有效成分葛根素进行定量测定.结果：葛根素在12～60 μg＆#183;ml-1内呈良好的线性关系（r =0.999 6）,平均加样回收率为100.10％,RSD为1.20％（n=6）.结论：该法简单方便,精密度高,准确可靠,可用于该制剂的质量控制.