黄芪抑制家兔心肌缺血再灌注时细胞凋亡的实验研究

目的：观察黄芪抑制家兔心肌缺血－再灌注时细胞凋亡的作用。方法：采用结扎冠状动脉左前降支后再通的方法,复制家兔心肌缺血－再灌注的动物模型,对心肌缺血－再灌注、黄芪治疗、心肌缺血、假手术等不同情况下心肌梗死面积（TTC法）、心肌细胞DNA电就凋亡细胞的形态（TUNEL法,电镜）进行了观察。结果：再灌注组和黄芪治疗组的梗死面积小于单纯缺血组;DNA电泳发现再灌注组呈凋亡的典型表现,单纯缺血组表现为细胞坏死,黄芪治疗组梯形条带中DNA含量较心肌缺血－再灌注组明显减少;TUNEL染色发现缺血组有散在阳性细胞,再灌注组有大量的阳性标记,且积聚成团,黄芪治疗组再灌注组明显减少;电镜观察发现再灌注组线粒体等细胞亚结构损伤严重,而黄芪治疗组则明显减轻。结论：缺血－再灌注可引起心肌细胞的凋亡,黄芪确实可抑制凋亡的发生。     

大鼠脑出血后神经元线粒体的体视学分析及黄芪的神经保护作用研究

目的　探讨黄芪对大鼠脑出血后神经元线粒体的保护作用。 方法　用Ⅶ型胶原酶脑内注入法制作大鼠脑出血模型,利用Narrow-alley Test检测各组大鼠的神经行为学指标;用透射电镜观察各组大鼠脑出血灶周围神经元线粒体的超微结构并对其进行体视学分析;用免疫组织化学方法检测Caspase-3蛋白表达。 结果　神经行为学检测,模型组大鼠的不对称分值明显升高,治疗组大鼠不对称分值较模型组明显下降;神经元线粒体体视学分析,与模型组相比,治疗组线粒体的体密度、数密度、比表面积和比膜面积均较模型组增大,差异有显著性（P＜0.05);与模型组比较,治疗组Caspase-3的阳性表达明显降低（P＜0.01)。 结论　黄芪能够减轻出血后神经元线粒体的损伤,抑制神经元凋亡,促进神经功能恢复。     

黄芪注射液对心肌细胞氧化应激性损伤的保护作用

目的探讨黄芪注射液(AI)对培养的心肌细胞氧化损伤的保护作用及其机制。方法体外培养的新生Wistar大鼠心肌细胞,分为对照组、 H₂O₂损伤组、药物治疗组： AI低、中、高（10、30、90 g/L）剂量组及AI(90 g/L)+L-NAME(20 μg/L)组。药物预先处理心肌细胞30 min后,加入H₂O₂损伤心肌细胞5 h,MTT法测定心肌细胞存活力,测定乳酸脱氢酶(LDH)和一氧化氮（NO）含量的变化;激光共聚焦检测心肌细胞活性氧（ROS）的变化;流式细胞仪检测心肌细胞线粒体膜电位及细胞凋亡率的变化。结果与H₂O₂损伤组比较,各剂量AI可提高细胞存活力 (P＜0.05),LDH漏出量降低(P＜0.01),活性氧荧光强度降低(P＜0.01),NO含量升高(P＜0.01),线粒体膜电位升高(P＜0.05),细胞凋亡率降低(P＜0.05)。与AI高剂量组比较,L-NAME组细胞存活力降低(P＜0.01),LDH漏出量升高(P＜0.01),活性氧荧光强度升高(P＜0.01), NO含量升高(P＜0.01),线粒体膜电位降低(P＜0.05),细胞凋亡率升高(P＜0.05)。结论黄芪注射液可对抗H₂O₂ 对心肌细胞的损伤,其机制与诱导NO生成、抑制活性氧生成引起的细胞凋亡有关。     

黄芪甲苷对肺纤维化大鼠的治疗作用及对肺组织中血清和糖皮质激素调节蛋白激酶1表达的影响

【目的】研究黄芪甲苷对肺纤维化（PF ）大鼠的治疗作用及对肺组织中血清和糖皮质激素调节蛋白激酶1（SGK1）表达的影响。【方法】36只SD大鼠,随机分为对照组、模型组和黄芪甲苷干预组。模型组和干预组气管内注射博来霉素（BLM ,5 mg／kg）诱导PF ,对照组在相同条件下给予生理盐水。干预组大鼠d2开始每天经胃管灌服0．1 g／L黄芪甲苷2 m L ,其余两组相同条件下给予生理盐水。治疗的d7和d28处死动物取出肺组织,进行病理学观察;用免疫组化和RT‐PCR检测各组鼠肺组织SGK1蛋白及mRNA表达的水平。【结果】病理学观察显示：与模型组比较,干预组肺泡炎和纤维化程度均减轻,SGK1蛋白及mRNA表达显著降低;与对照组比较,干预组存在一定程度肺泡炎和纤维化,SGK1蛋白及mRNA表达明显高于对照组。【结论】黄芪甲苷可能通过抑制SGK1的过度表达,对实验性大鼠PF具有良好的治疗作用。     

阿维A联合黄芪治疗银屑病临床疗效研究及对TNF-α的影响

目的 观察阿维A联合黄芪注射液治疗银屑病的临床疗效及血清中TNF-α水平的变化,探讨阿维A联合黄芪注射液治疗银屑病的临床疗效和肿瘤坏死因子(TNF-α)与银屑病的关系.方法 临床随机分为治疗组11例和对照组9例,另设20例健康人做空白对照观察.对照组口服阿维A 0.2～0.6 mg/kg·d,皮损好转后再逐渐减量维持,直至临床治愈;治疗组在对照组治疗基础上静脉注射黄芪注射液.检测治疗前后血清中TNF-α水平,并与空白对照组比较.结果 治疗前治疗组与对照组血清中TNF-α的水平分别为(45.4±12.4)ng/L和(46.4±16.9)ng/L,治疗后分别为(13.8±5.4)ng/L和(20.9±6.3)ng/L,同组治疗前后比较差异有统计学意义,治疗组治疗后与对照组比较,差异有统计学意义.结论 阿维A联合黄芪治疗银屑病疗效较好,可能通过抑制TNF-α发挥抗炎及免疫调节作用.     

参附、黄芪注射液辅助治疗原发性肾病综合征的疗效观察

目的:观察参附、黄芪注射液辅助治疗原发性肾病综合征(PNS)的效果。方法:47例患者随机分为治疗组和对照组,在常规治疗的同时,两组均给予强的松1mg.kg-1.d-1)剂量晨顿服。治疗组在次基础上加用黄芪注射液40m l,参附注射液50m l,1次/d,共用28d。通过检测两组治疗前后24h尿蛋白变化来观察临床治疗效果。结果:治疗组与对照组相比,总有效率差异有统计学意义(P<0.05)。结论:参附、黄芪注射液辅助激素治疗原发性肾病综合征疗效较好。     

3,5-二硝基水杨酸比色法测定黄芪多糖的含量

目的：建立黄芪有效部位多糖的含量测定方法。方法：用优化后的3,5-二硝基水杨酸（DNS）比色法测定黄芪有效部位多糖的含量。结果：DNS试剂的用量为2ml,显色温度为100℃,显色时间为5min,在500nm处多糖含量与吸光度有良好的线性关系,平均回收率为100.6%,RSD=1.79%（n=5）。结论：该方法操作简便,测定结果准确,重复性好,可用于测定黄芪多糖的含量。     

Astragalus membranaceus flavonoids (AMF) ameliorate chronic fatigue syndrome induced by food intake restriction plus forced swimming

Aim of the study

Alteration of immune function may be associated with chronic fatigue syndrome (CFS) and this study reveals the immunoregulatory effect of Astragalus membranaceus flavonoids (AMF).

Materials and methods

CF rats were induced by food intake restriction plus forced swimming for 6 weeks.

Results

An atrophied spleen associated with a significantly decreased spleen/body weight ratio and a reduced spleen cells proliferation was found in CF rats when compared with home cage controls. AMF given orally at 20, 50 and 100 mg/kg body weight once a day consecutively for 6 weeks could recover the reduced cell proliferation. A switch to Th1-dominated immune regulation was observed in CF rats as the cultured splenocytes produced more interleukin-2 (IL-2) but less IL-4 when compared with controls. Supplementation with AMF could significantly counteract the aberrant cytokine production and rats received AMF exhibited higher endurance capacity to swim when compared with those without AMF administration. Checking the spectrum signals confirmed that the three major isoflavones contained in AMF were ononin, formononetin, and demethylhomopterocarpin.

Conclusion

Alterations of immune function may be associated with CFS and the tonic effects of AMF against CF may be attributable to balance the abnormal cytokine level by isoflavones.     

Induction of apoptosis in human hepatocarcinoma SMMC-7721 cells in vitro by flavonoids from Astragalus complanatus

Aim of the study

Flavonoids extracted from the seeds of Astragalus complanatus R.Br. reduce the proliferation of many cancer cells. The present study was carried out to evaluate the effects of these flavonoids from Astragalus complanatus (FAC) on human hepatocarcinoma cell viability and apoptosis and to investigate its mechanisms of action in SMMC-7721 cells.

Materials and methods

Cell viability was measured using the MTT assay. To detect apoptotic cells, SMMC-7721 cells treated with FAC were stained with Hoechst 33258 and subjected to agarose gel electrophoresis. Quantitative detection of apoptotic cells was performed by flow cytometry. The effects of FAC on apoptosis and cell cycle regulatory genes and proteins in SMMC-7721 cells were examined using an S series apoptosis and cell cycle gene array and Western blot analysis.

Results

The growth of SMMC-7721 and HepG2 cells was inhibited by treatment with FAC. Cell death induced by FAC was characterized by nuclear condensation and DNA fragmentation. Moreover, the cell cycle was arrested in the G0/G1 and S phases in FAC-treated SMMC-7721 cells. A sub-G1 peak with reduced DNA content was also formed. The activity of caspase-3 was significantly increased following FAC treatment. Microarray data indicated that the expression levels of 76 genes were changed in SMMC-7721 cells treated with FAC: 35 genes were up-regulated and 41 were down-regulated. Western blot analysis showed that caspase-3, caspase-8, Bax, P21, and P27 protein levels in SMMC-7721 cells were increased after 48 h of FAC treatment, while cyclinB1, cyclinD1, CDK1, and CDK4 protein levels were decreased.

Conclusions

These results suggest that FAC may play an important role in tumor growth suppression by inducing apoptosis in human hepatocarcinoma cells via mitochondria-dependent and death receptor-dependent apoptotic pathways.     

黄芪对糖尿病肾病大鼠肾组织COX-2和NF-κB表达的影响

目的：探讨环氧合酶-2（COX-2）和核转录因子κB（NF-κB）在糖尿病肾病大鼠肾组织中的表达及黄芪对其的影响。方法：将大鼠分成正常对照组、糖尿病肾病组、黄芪干预组。12周周末检测血肌酐、尿素氮、尿蛋白定量;应用免疫组化、Western印迹和聚合酶链式反应方法分别检测大鼠肾组织COX-2和NF-κB的表达。结果：与正常对照组相比,糖尿病肾病组大鼠COX-2和NF-κB的基因及蛋白表达升高（均P〈0.01）。黄芪干预组大鼠肾组织COX-2和NF-κB的基因及蛋白表达较糖尿病肾病组下调（均P〈0.01）。结论：COX-2参与了糖尿病肾病发生发展的病理过程,黄芪治疗糖尿病肾病的作用机制之一是通过下调COX-2的表达从而发挥肾脏保护作用。