大承气汤治疗大鼠内毒素性ARDS的疗效分析及免疫调节机制研究

目的： 探讨大承气汤对内毒素“二次打击”致急性呼吸窘迫综合征（ARDS）的防治作用,为中西医结合防治呼吸道疾病提供有效依据。方法: 48只健康雄性Wistar大鼠随机分为4组:生理盐水对照组、ARDS模型组、大承气汤治疗组、地塞米松治疗组,每组12只。采用大肠杆菌脂多糖(LPS) “二次打击”建立大鼠ARDS动物模型,结合动脉血气分析、肺湿/干重(W／D)比值及肺组织病理学观察和评分,评价大承气汤的药理作用;酶联免疫吸附法（ELISA）测定血浆与支气管肺泡灌洗液（BALF）中肿瘤坏死因子-α（TNF-α）、白细胞介素-1（IL-1）、IL-10水平,探讨大承气汤的药理作用和免疫调节机制。结果: (1)大承气汤可以显著提高ARDS大鼠动脉血氧分压,增加血氧饱和度,降低肺湿/干重比值和肺病理损伤,减轻肺水肿和肺部炎症反应,因而具有改善肺通气、抑制肺部炎症反应、减轻肺损伤功能。(2)大承气汤可使血浆促炎细胞因子（TNF-α、IL-1）和抗炎细胞因子（IL-10）水平同时降低,使BALF中TNF-α、IL-1水平降低同时IL-10水平却升高,因而表现出对全身炎症反应和局部炎症反应不同的免疫调节机能。结论: 大承气汤抑制肺部炎症反应除与下调全身炎症反应水平有关,同时还与促进肺部抗炎介质产生、调节肺部促炎介质/抗炎介质的平衡有关。     

黄连解毒汤干预热病证候的代谢组学研究

目的:研究黄连解毒汤对热病证候整体代谢的影响,探讨其干预热病证候的作用机制。方法:应用代谢组学技术,利用超高液相-四极杆-飞行时间/质谱(UPLC-Q-TOF/MS)检测分析系统,采用主成分分析(PCA)和偏最小二乘-判别分析(PLS-DA)统计分析方法,研究黄连解毒汤对2,4-二硝基苯酚和干酵母诱导的热病证候的干预作用。结果:两个模型在同一得分图中呈现明显的聚类分组,说明两个模型所致机体代谢的改变不同;分别给予黄连解毒汤干预后,两个模型的各实验组代谢物组在得分图中聚类于模型恢复的路线上,表明黄连解毒汤能加快热病证候的恢复并使其向着健康的方向发展。结论:基于代谢组学整体表征了黄连解毒汤对不同原因诱导的热病证候具有良好的治疗作用。     

胃肠宁对利血平所致大鼠脾虚模型的影响

目的 观察胃肠宁对大鼠脾虚证模型的影响.方法 36只大鼠随机分为正常对照组、脾虚模型组、四君子汤组和胃肠宁组,每组9只.采用利血平复制大鼠脾虚模型,连续灌胃14 d后,处死大鼠,分离肝、十二指肠和血清,测定各组大鼠血浆LDH、AKP、AchE、CK和肝组织匀浆中SOD、MDA、GSH-Px、NOS含量变化,并观察各组大鼠肝和十二指肠病理变化.结果 与正常对照组相比,脾虚模型组大鼠LDH、AchE、CK、SOD、GSH-Px、NOS含量明显降低,而AKP、MDA含量明显升高;与脾虚模型组相比,胃肠宁组LDH、AchE、CK、SOD、GSH-Px、NOS含量显著升高,而AKP、MDA含量显著降低;脾虚模型组大鼠肝组织发生广泛性颗粒变性和空泡变性;十二指肠上皮及绒毛明显脱落,大量炎性细胞浸润,杯状细胞数量明显减少,胃肠宁组与脾虚组相比,肝和十二指肠病理变化明显减轻.结论 胃肠宁对利血平所致大鼠脾虚具有较好的治疗作用.     

围绝经期妇女重度抑郁症综合康复治疗的临床疗效分析

目的探讨围绝经期妇女重度抑郁症综合康复治疗的临床疗效。方法对26例围绝经期重度抑郁症妇女给予服用尼尔雌醇、氟西汀、解郁汤及心理治疗,采用汉密尔顿抑郁量表（HAMD）和自评抑郁量表（SDS）评分进行治疗前后对比观察和治疗效果评价,同时检测血清雌二醇（E2）、血清促卵泡素（FSH）含量变化。结果经过8周的综合康复治疗后,患者E2水平明显增高（P〈0．01）,FSH含量显著降低（P〈0．01）,HAMD和SDS评分明显降低（P〈0．01）。26例患者经治疗后,治愈16例（61．5％）,显效6例（23．1％）,有效4例（15．4％）,总有效率为100％。结论综合康复治疗围绝经期妇女重度抑郁症疗效好、副作用小,是安全有效的治疗方法。     

当归补血汤对去势大鼠血清SOD、MDA含量和脾脏、子宫指数的影响

目的:研究当归补血汤对去势大鼠血清超氧化物歧化酶(SOD)、丙二醛(MDA)含量和脾脏、子宫指数的影响。方法:实验分为5组,即假手术、模型、六味地黄丸和当归补血汤高、低浓度组。将3月龄SD未孕雌性大鼠给予单侧切除卵巢造模,酶联免疫法测定大鼠血清SOD、MDA含量和脾脏、子宫指数。结果:与模型组比较,当归补血汤高、低浓度和六味地黄丸组血清MDA含量均显著降低(P<0.01),血清SOD活性和脾脏、子宫指数均显著升高(P<0.01)。结论:当归补血汤抗衰老和增强抵抗力的内在机制之一可能与提高清除自由基能力以及增加免疫器官重量有关,高浓度当归补血汤作用更为明显。     

Neuroprotective effects of Buyang Huanwu decoction against hydrogen peroxide induced oxidative injury in Schwann cells

Ethnopharmacological relevance

Buyang Huanwu decoction (BYHWD) is a traditional Chinese medicine and can be used to promote peripheral nerve regeneration. However the regenerative mechanism of BYHWD remains unclear. The objective of this study was to investigate the protective mechanisms of BYHWD in Schwann cells damaged by hydrogen peroxide (H₂O₂).

Materials and methods

Schwann cells which were derived from neonatal sciatic nerves of rats were used in subsequent experiments. Schwann cells were injured by various concentrations of H₂O₂ (0.25, 0.5 and 1 mM final concentration). BYHWD (600 μg/ml final concentration) was added to the medium either simultaneously or 1 h later after the addition of H₂O₂. Subsequently, methyl thiazolyl tetrazolium (MTT) assay was performed. Superoxide dismutase (SOD) activity and malondialdehyde (MDA) levels were also examined after 12 h. The expression of Caspase 3 and the concentration of intercellular Ca²⁺ ([Ca²⁺]i) were also determined.

Results

Among three concentrations of H₂O₂, 0.5 mM H₂O₂ induced Schwann cells swelled and neuritis disappeared after 12 h. In the presence of BYHWD, MTT assay showed that more cells were viable in comparison with the H₂O₂ injury group. Moreover, the addition of BYHWD has also increased the SOD activity with decreased in MDA level. Furthermore, the concentration of [Ca²⁺]i and expression of Caspase 3 were decreased with the addition of BYHWD in culture.

Conclusions

Our results revealed that BYHWD protected Schwann cells from oxidative injury. The mechanism of BYHWD promoting neural regeneration possibly associated with its anti-oxidative activity.     

Danshen-Gegen decoction exerts proliferative effect on rat cardiac myoblasts H9c2 via MAPK and insulin pathways

Ethnopharmacological relevance

Danshen (root of Salvia miltiorrhiza) and Gegen (roots of Pueraria lobata) are traditional Chinese medicines that have been used in combination for cardiovascular disease treatment.

Aim of the study

The present study was performed to investigate the effect of Danshen-Gegen decoction on rat myocardium cell line H9c2 and the possible molecular mechanisms.

Materials and methods

Rat heart myocardium H9c2 cells were treated with or without Danshen-Gegen decoction (DG) ranging from 10 to 1000 μg/ml for 24 h. Cell viability was measured by Alarma blue assay and cell proliferation assay was performed by BrdU Cell Proliferation ELISA kit. The activation of mitogen-activated protein kinase and insulin pathways was analyzed by Luminex technology and the growth factors and cytokine expression of H9c2 cells induced by DG was evaluated by protein array. Moreover, a rat functional specific cDNA microarray was constructed to study the gene expression profiles of H9c2 cells upon the DG treatment at 50 μg/ml for 24 h.

Results

DG promoted H9c2 cell viability and cell proliferation at dose-dependent manner within the range between 0 and 250 μg/ml. A Bio-Plex assay kit (Bio-Rad Bioscience) was used to detect the expression level of phosphoprotein as well as total proteins involved in the MAPK and insulin pathways. Significant phosphorylation of ERK, c-Jun, JNK, p38, AKT, IGF-IR, IRS-1and I kappa B were observed after DG treatment at 2 h or 4 h. A rat cytokine antibody array was used to detect and quantify 22 growth factors and cytokines in samples collected from the control and DG treated H9c2 cells. In the category of growth factors, GM-CSF, CNIF and b-NGF were stimulated by DG, while the expression of TIMP-1 was suppressed. For cytokine expression, it was found that DG stimulated three interleukin subclasses, IL-1α, 1X and 6, respectively. However, the expression of pro-inflammatory factors such as TNF-α and IFN-γ were down-regulated significantly. Moreover, the microarray analysis revealed that DG significantly up-regulated anti-apoptosis related genes such as Cdkn2c and Ppp3ca, and several cardiovascular disease suppressers and anti-inflammatory mediators; on the other hand, pro-apoptotic related genes including Caspase and Tnf-α were down-regulated by DG. Based on the results, a tentative scheme was proposed to show that the activation of the MAPK and insulin pathways are involved in the bioactive effect of Danshen-Gegen decoction on cardiomyocytes.

Conclusion

Our study suggested that Danshen-Gegen decoction has proliferative effect on myocardium cells via MAPK and insulin signaling pathways. The molecular mechanism of the action may include the up-regulation of IRS/AKT and JNK pathways as well as the inhibition of TNF and p38 pathways.     

Effects of food and gender on the pharmacokinetics of rhein and emodin in rats after oral dosing with Da‐Cheng‐Qi decoction

Da‐Cheng‐Qi decoction (DCQD), a traditional Chinese medicine preparation used to treat digestive diseases, is composed of dahuang (Rheum officinale Baill, Polygonaceae), houpu (Magnolia officinalis Rehd., Magnoliaceae), zhishi (Citrus aurantium L, Rutaceae) and mangxiao (sodium sulphate). Rhein and emodin are the major active components of Rheum officinale Baill. To investigate the effects of food and gender on the plasma concentrations of rhein and emodin after oral administration of DCQD, a rapid high‐performance liquid chromatographic method was developed and validated. A reversed phase C₁₈ column (150 × 4.6 mm) and a mobile phase of methanol and 0.2% acetic acid (78:22, v/v) were employed with ultraviolet detection at 254 nm. Feeding was observed to decrease the absorption of rhein and emodin in rats receiving DCQD orally. No evidence for gender‐based differences in the pharmacokinetics of rhein was observed. However, the half‐life and area under the concentration–time curve for emodin differed significantly between male and female rats. Because food intake and gender are anticipated to influence the pharmacokinetics of DCQD in human subjects, it is recommended that oral doses of DCQD be reduced in fasting subjects and that female patients receive lower oral doses compared with male patients. Copyright © 2010 John Wiley & Sons, Ltd.     

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目的    探讨生芦润燥汤治疗干燥综合征的临床疗效。方法    选取2007年5月至2008年12月上海中医药大学附属市中医医院风湿科及口腔科门诊收治的60例干燥综合征患者,随机分为试验组和对照组,每组30例。试验组服用生芦润燥汤治疗,对照组服用硫酸羟氯喹治疗,两组疗程均为12个月。治疗期间每3个月随访1次,观察并记录两组患者治疗前后的症状、体征及实验室检查结果。结果    试验组疗效与对照组疗效有显著差异（P < 0.01）;在改善症状、体征和中医征候方面,试验组明显优于对照组（P < 0.01）。结论    生芦润燥汤治疗干燥综合征有显著疗效。