3′-Daidzein sulfonate sodium protects against memory impairment and hippocampal damage caused by chronic cerebral hypoperfusion

3′-Daidzein sulfonate sodium（DSS） is a new synthetic water-soluble compound derived from daidzein,a soya isoflavone that plays regulatory roles in neurobiology.In this study,we hypothesized that the regulatory role of DSS in neurobiology exhibits therapeutic effects on hippocampal damage and memory impairment.To validate this hypothesis,we established rat models of chronic cerebral hypoperfusion（CCH） by the permanent occlusion of the common carotid arteries using the two-vessel occlusion method.Three weeks after modeling,rat models were intragastrically administered 0.1,0.2,and 0.4 mg/kg DSS,once a day,for 5 successive weeks.The Morris water maze test was performed to investigate CCH-induced learning and memory deficits.TUNEL assay was used to analyze apoptosis in the hippocampal CA1,CA3 regions and dentate gyrus.Hematoxylin-eosin staining was performed to observe the morphology of neurons in the hippocampal CA1,CA3 regions and dentate gyrus.Western blot analysis was performed to investigate the phosphorylation of PKA,ERK1/2 and CREB in the hippocampal PKA/ERK1/2/CREB signaling pathway.Results showed that DSS treatment greatly improved the learning and memory deficits of rats with CCH,reduced apoptosis of neurons in the hippocampal CA1,CA3 regions and dentate gyrus,and increased the phosphorylation of PKA,ERK1/2,and CREB in the hippocampus.These findings suggest that DSS protects against CCH-induced memory impairment and hippocampal damage possibly through activating the PKA/ERK1/2/CREB signaling pathway.

3′-Daidzein sulfonate sodium improves mitochondrial functions after cerebral ischemia/reperfusion injury

3′-Daidzein sulfonate sodium is a new synthetic water-soluble compound derived from daidzein（an active ingredient of the kudzu vine root）. It has been shown to have a protective effect on cerebral ischemia/reperfusion injury in rats. We plan to study the mechanism of its protective effect. 3′-Daidzein sulfonate sodium was injected in rats after cerebral ischemia/reperfusion injury. Results showed that 3′-daidzein sulfonate sodium significantly reduced mitochondrial swelling, significantly elevated the mitochondrial membrane potential, increased mitochondrial superoxide dismutase and glutathione peroxidase activities, and decreased mitochondrial malondialdehyde levels. 3′-Daidzein sulfonate sodium improved the structural integrity of the blood-brain barrier and reduced blood-brain barrier permeability. These findings confirmed that 3′-daidzein sulfonate sodium has a protective effect on mitochondrial functions after cerebral ischemia/reperfusion injury, improves brain energy metabolism, and provides protection against blood-brain barrier damage.

Protective effects of 3′-daidzein sulfonate sodium on chronic hepatic injury by regulating the T lymphocyte subsets in mice

OBJECTIVE To explore the protective effects of 3′-daidzein sulfonate sodium on chronic hepatic injury induced by carbon tetrachloride(CCl4)in mice and investigate the mechanism about regulating the T lymphocyte subsets.METHODS Healthy Kunming male mice were randomly divided in 5groups:control group,model group,bifendate positive control group(2.5mg·kg-1),low and high dose 3′-daidzein sulfonate sodium groups(0.1and 0.3mg·kg-1).The chronic hepatic injury mice were made by intraperitoneal injection of 10% CCl4 plant oil solution twice a week,and sustained for six weeks.At the same time,the mice were treated with normal saline,bifendate(2.5mg·kg-1)and 3′-daidzein sulfonate sodium(0.1and 0.3mg·kg-1),respectively by ig administration once a day and continued for six weeks.After the last administration,the mice blood and liver were taken.Using automatic biochemical analyzer survey the activity of aspartate aminotransferase(AST)and alanine aminotransferase(ALT)in serum.Flow cytometry was used to detect T lymphocyte subsets,enzymes analysis technique was used to observe the liver function(CD3+,CD4+and CD8+),HemateinEosin stain was used to explore the changes in liver morphology.RESULTS The level of ALT and AST in the serum in model group mice increased significantly.While the level of ALT and AST in the serum in 3′-daidzein sulfonate sodium(0.1 or 0.3mg·kg-1,ig)groups mice decreased compared with the model group(P<0.05).And compared with the model group,the ratios of CD3+,CD4-CD8-and CD4+/CD8+ decreased,and the ratios of CD8+ increased in 3′-daidzein sulfonate sodium groups(0.1 and 0.3mg·kg-1,ig;P<0.05).CONCLUSION 3′-daidzein sulfonate sodium have a protective effect on CCl4-induced liver injury mice,and it can result in the changes of T lymphocyte subsets,which may be one of the factors leading to hepatic injury by CCl4.

DSS对脑缺血再灌注损伤的保护与抗氧化作用的关系

目的研究3′-大豆苷元磺酸钠(DSS)对脑缺血再灌注损伤的保护作用,并初步探讨其保护机制与抗氧化作用的关系。方法大鼠大脑中动脉栓塞,制作局灶性脑缺血再灌注损伤模型,缺血后10min,于舌下静脉给不同剂量的DSS。再灌注24h后,腹腔静脉取血并分离血清,断头取脑,一组前脑做TTC染色,测定脑梗死体积;另一组左脑制成10%的脑组织匀浆。比色法测血清及脑组织匀浆SOD、LDH、GSH-Px的活性及MDA的含量。结果与模型组相比,各剂量的DSS治疗组脑梗死体积减小;血清LDH活性及MDA含量均降低;脑组织SOD、GSH-Px的活性显著升高。结论DSS对脑缺血再灌注损伤具有一定的保护作用,其保护机制与DSS提高抗氧化作用、增强氧自由基的清除有关。

DSS对大鼠脑缺血再灌注损伤后视网膜作用的影响

目的:研究3′-大豆苷元磺酸钠(DSS)对大鼠视网膜缺血再灌注损伤的影响。方法:用夹闭大鼠一侧颈总动脉的方法进行动物建模,制备大鼠视网膜缺血模型。随机分成3组:生理盐水组、不同剂量DSS治疗组(1、2mg/kg)。在夹闭大鼠一侧颈总动脉前分别舌下静脉注射相DSS或生理盐水。在夹闭大鼠一侧颈总动脉前、后1h及灌注后1h分别行眼底荧光血管造影(FFA)和视网膜电流图(ERG)检查。结果:与对照组相比,再灌注后1h与缺血前比较,DSS治疗组ERG振幅具有显著性差异。各组荧光血管造影(FFA)显示,DSS组视网膜血管痉挛明显恢复,静脉血管不规则扩张和迂曲;FFA显示DSS组灌注后视网膜动脉充盈时间明显缩短,且具有一定的剂量依赖性。结论:DSS能扩张视网膜血管,对视网膜缺血再灌注损伤有明显的保护作用。

3′-大豆苷元磺酸钠对离体心肌缺血再灌注时NO ANG Ⅱ及ANP水平的影响

目的:研究3′-大豆苷元磺酸钠(DSS)对离体心脏缺血再灌注损伤的保护作用及NO、ANG Ⅱ和ANP含量的变化。方法:应用大鼠离体心肌缺血再灌注损伤模型,在灌流液中加入高、中、低剂量的DSS。测定冠脉流量、左心室收缩压及心肌组织NO、ANG Ⅱ和ANP含量的变化。结果:各剂量的3′-大豆苷元磺酸钠均能有效地增加冠脉流量,增强左心室收缩压;三种剂量的DSS均能升高心肌组织NO含量(P<0.05);高剂量的DSS使心肌组织ANG Ⅱ(P<0.05)含量降低,低剂量的DSS使ANP含量升高(P<0.05)。结论:DSS可通过增加NO、ANP的合成,减少ANG Ⅱ的释放,扩张冠脉,增强心肌收缩力,从而对离体心脏缺血再灌注损伤起保护作用。

3'-大豆苷元磺酸钠对脑缺血再灌注损伤大鼠学习记忆及Arc表达的影响

目的:研究3'-大豆苷元磺酸钠(DSS)对局灶性脑缺血再灌注损伤后大鼠的学习记忆及海马脑组织活性调节细胞骨架蛋白(Arc)表达的影响。方法:采用大鼠大脑中动脉栓塞制作局灶性脑缺血再灌注(MCAO)模型,实验随机分为假手术组、MCAO再灌注模型组、MCAO再灌注模型+3'-大豆苷元磺酸钠组(2 mg·kg-1DSS组)。给予DSS连续灌胃治疗28天,采用Morris水迷宫行为学(定位航行实验及空间探索实验)观测各组实验动物空间学习记忆能力,western-blot法检测大鼠海马脑组织Arc的表达变化。结果:Morris水迷宫定位航行实验中,与模型组相比,2 mg·kg-1DSS组逃避潜伏期明显缩短;空间探索实验中,3'-大豆苷元磺酸钠给药组在第Ⅲ象限(原平台所在象限)游泳路程与总游泳路程的比值(sⅢ/s T)明显高于模型组;western-blot法检测结果显示,与假手术组比较,模型组大鼠Arc表达明显下降;与模型组比较,2 mg·kg-1DSS组Arc的表达明显增高。结论:3'-大豆苷元磺酸钠可能通过上调细胞骨架蛋白(Arc)的表达来改善缺血再灌注损伤大鼠的学习记忆功能。

3'-大豆苷元磺酸钠对脑缺血/再灌注损伤大鼠血脑屏障结构及通透性的影响

目的：观察3＇-大豆苷元磺酸钠对脑缺血／再灌注损伤大鼠血脑屏障结构及通透性的影响。方法：采用MCAO方法制备脑缺血再灌注大鼠模型，并通过尾静脉给予3＇-大豆苷元磺酸钠治疗，采用静脉注射伊文思蓝观察血脑屏障通透性的改变，采用透射电镜观察血脑屏障超微结构的改变。结果：3＇-大豆苷元磺酸钠可显著改善血脑屏障通透性，表现为降低伊文思蓝在脑中的分布，血脑屏障超微结构显示3＇-大豆苷元磺酸钠可显著提高血脑屏障的完整性，能显著减轻血管周围星型胶质细胞和周细胞的肿胀，保持血管的正常结构。结论：3＇-大豆苷元磺酸钠在0．5～2．0 mg·kg^－1范围内可显著保护脑缺血／再灌注损伤大鼠引发的血脑屏障损伤。

3'-大豆苷元磺酸钠对前列腺癌细胞的影响

目的:通过观察3'-大豆苷元磺酸钠对前列腺癌细胞增殖和相关基因表达的影响。方法:将前列腺癌细胞在培养基(含10%小牛血清)中采用开放式单层贴壁培养,采用MTT法对前列腺癌细胞的增殖情况进行分析,对3'-大豆苷元磺酸钠作用后的雄激素非依赖型前列腺癌细胞的存活率、细胞毒作用进行了研究。结果:与对照组相比,3'-大豆苷元磺酸钠对前列腺癌细胞抑制效应具有剂量依赖性,具有诱导前列腺癌细胞凋亡和引起坏死效应,同时诱导前列腺癌细胞的(PTEN基因)表达。结论:3'-大豆苷元磺酸钠具有明显抑制前列腺癌细胞增殖效应。

3′-大豆苷元磺酸钠对四氯化碳所致小鼠肝损伤和肝药酶的影响

目的研究3′-大豆苷元磺酸钠(3′-DSS)对四氯化碳(CCl4)所致小鼠肝损伤的影响。方法取雄性昆明种小鼠60只,每组12只,随机分为对照组、模型组、联苯双酯阳性对照组、0.1 mg/kg 3′-DSS组和0.3 mg/kg 3′-DSS组。每星期进行2次腹腔注射10%CCl4植物油溶液0.1 ml/10 g,造成慢性肝损伤模型,期间给予灌服2.5 mg/kg联苯双酯、0.1 mg/kg 3′-DSS、0.3 mg/kg 3′-DSS,持续6周,给药结束后,分别取血和肝脏,分离血清,制备肝均浆,检测丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)的活性和肝药酶含量。结果与对照组比较,模型组小鼠肝功能明显下降,ALT、AST水平明显升高,肝药酶含量明显下降;与模型组比较,0.1、0.3 mg/kg 3′-DSS治疗组ALT、AST水平均有明显下降;肝药酶含量也明显增加。结论 3′-DSS对CCl4所致小鼠肝损伤具有明显的保护作用。