Panax notoginseng saponins suppress oxygen-glucose deprivation/reoxygenation-induced pyroptosis of SH-SY5Y cells

AIM To investigate the effect of Panax notoginseng saponins (PNS) on pyroptosis of SH-SY5Y cells induced by oxygen-glucose deprivation/reoxygenation (OGD/R). METHODS The OGD/R was conducted to induce ischemia/reperfusion injury in SH-SY5Y cells. The effects of PNS on the viability (detected by CCK-8 assay) and membrane permeability [indicated by lactate dehydrogenase (LDH) leakage and propidium iodide (PI) staining positive cell proportion] of OGD/R-induced SH-SY5Y cells were observed. The protein levels of gasdermin D (GSDMD), GSDMD N-terminal fragment (GSDMD-N), caspase-1 and caspase-4, and the release of interleukin-1β (IL-1β) and IL-18 in the cells were also determined. RESULTS After exposure to OGD/R, the viability of SH-SY5Y cells dramatically decreased (P<0.01), while the LDH leakage, the PI staining positive cell proportion, the protein levels of GSDMD, GSDMD-N, caspase-1 and caspase-4, and the release of IL-1β and IL-18 were significantly increased (P<0.01). However, PNS treatment enhanced the viability of SH-SY5Y cells inhibited by OGD/R (P<0.01), but reduced the leakage of LDH and the percentage of PI staining positive cells (P<0.05 or P<0.01). Moreover, PNS reversed the increases in the protein levels of GSDMD, GSDMD-N, caspase-1 and caspase-4 and the release of IL-1β and IL-18 in OGD/R-induced SH-SY5Y cells (P<0.05 or P<0.01). CONCLUSION Treatment with PNS alleviates OGD/R-induced injury in SH-SY5Y cells. Its mechanism may be related to inhibition of SH-SY5Y cell pyroptosis induced by OGD/R.     

苜蓿生物活性物质在动物上的应用

苜蓿中含有许多生物活性物质,能够显著影响动物的生长性能、繁殖机能和免疫功能,有些活性物质还可以作为人的保健品使用。本文主要介绍苜蓿皂苷、苜蓿多糖、苜蓿黄酮的生物学功能以及国内外学者对苜蓿中活性成分研究的最新成果。苜蓿中活性物质可以作为动物的饲料添加剂使用,一定剂量的苜蓿活性物质可以提高动物体内的淋巴细胞水平,增加免疫力;提高谷胱甘肽过氧化物酶的活性,抑制丙二醛的水平,具有抗氧化功能;增加动物的生产性能和繁殖性能。     

New furostanol saponins from Smilax aspera L. and their in vitro cytotoxicity

The occurrence of the two new cis-fused A/B rings furostanol saponins (25S)-26-O-β-d-glucopyranosyl-5β-furostan-1β,3β,22α,26-tetraol-1-O-β-d-glucopyranoside and (25S)-26-O-β-d-glucopyranosyl-5β-furostan-1β,2β,3β,5β,22α,26-hexaol and the known compounds (25S)-26-O-β-d-glucopyranosyl-5β-furostan-3β,22α,26-triol-3-O-α-l-rhamnopyranosyl-(1→2)-O-β-d-glucopyranosyl-(1→2)-O-β-d-glucopyranoside and (25S)-26-O-β-d-glucopyranosyl-5β-furostan-3β,22α,26-triol-3-O-β-d-glucopyranosyl-(1→2)-O-β-d-glucopyranoside, trans-resveratrol, (+) catechin and (−) epicatechin in the rhizomes of Smilax aspera is reported. All saponins have been isolated as their 22-OMe derivatives, which were further subjected to extensive spectroscopic analysis. The isolated furostanol saponins were evaluated for cytotoxic activity against human normal amniotic and human lung carcinoma cell lines using neutral red and MTT assays. In vitro experiments showed significant cytotoxicity in a dose dependent manner with IC₅₀ values in the range of 32.98-94.53 µM.     

大豆多肽对西洋参生物生长量与人参皂苷Rb1含量的影响

研究大豆多肽对西洋参(Panax quinquefoliusL.)生物生长量和人参皂苷Rb1含量的影响。将大豆多肽2号粉和3号粉分别配制成高、中、低3种浓度的水溶液,以叶面喷洒与根部灌注的方式,对生长3年的西洋参进行实验;同时观察生物生长量和人参皂苷Rb1的变化并总结规律。大豆多肽2号粉4mg/ml浓度根部灌注对西洋参生物生长量与人参皂苷Rb1增加作用最明显。研究结果为提高西洋参产量与质量提供理论依据,在西洋参的栽培种植中起到了实践指导作用。     

紫花苜蓿MsSQE1的克隆及对皂甙合成的功能分析

【目的】鲨烯环氧酶（squalene epoxidases,SQE）是苜蓿皂甙合成途径中的一种限速酶,与苜蓿皂甙的合成密切相关。通过对苜蓿鲨烯环氧酶（MsSQE1）基因的克隆及在苜蓿中过表达,探究鲨烯环氧酶对皂甙合成的作用机制。【方法】以模式植物蒺藜苜蓿鲨烯环氧酶基因序列设计引物,同源克隆苜蓿MsSQE1。对MsSQE1进行生物信息学分析;通过基因枪轰击技术,使MsSQE1在洋葱表皮瞬时表达,进行亚细胞定位。利用qRT-PCR方法分析该基因在根、茎、叶中的表达水平,以及在紫外辐射、ABA和GA₃条件下的表达模式。在茉莉酸甲酯（MeJA）的诱导下,分析MsSQE1的转录水平,及对苜蓿皂甙含量的影响。利用根癌农杆菌转化体系,获得过表达MsSQE1的阳性转基因植株,并测定转基因植株的皂甙含量。【结果】克隆了MsSQE1的cDNA序列,开放阅读框1 578 bp,编码525个氨基酸,等电点为8.59。同源性比对分析,其氨基酸序列与蒺藜苜蓿中SQE1氨基酸序列同源性为98.6%,与拟南芥的同源性为80%。亚细胞定位显示,MsSQE1可能定位于细胞膜。组织特异性表达分析显示,MsSQE1在叶中的表达量最高,茎中次之,根中的表达量最低。在紫外辐射、ABA和GA₃的诱导表达显示,紫外辐射诱导24 h,叶中表达量最高; GA₃（50 μmol·L ^-1）和 ABA（100 μmol·L ^-1）处理,均为8 h叶中表达量最高;MeJA处理能诱导MsSQE1表达量上调的同时苜蓿总皂甙含量也随着MsSQE1表达的上调而增加。分析过表达MsSQE1转基因苜蓿和转空载体苜蓿株系发现,MsSQE1的表达水平和总皂甙含量均升高,其中MsSQE1的表达水平是对照的3.11—9.45倍,总皂甙含量比对照提高14.26%—28.05%,暗示MsSQE1是苜蓿皂甙合成途径中的一种关键调节酶。【结论】从豆科牧草紫花苜蓿中克隆了MsSQE1并进行功能分析。在苜蓿中过表达MsSQE1能增加苜蓿总皂甙含量,暗示MsSQE1的表达影响苜蓿皂甙的生物合成,可能对皂甙的合成有重要的调节作用。     

Evaluation of Agave lechuguilla by‐product crude extract as a feed additive for juvenile shrimp Litopenaeus vannamei

Agave lechuguilla is a succulent plant species, mainly distributed in the northeast of Mexico and south of the United States of America. The main use for this plant is the fibre´s extraction (known as Tampico fibres), resulting in 15% of fibres and 85% of a by‐product waste named guishe. The lechuguilla collectors, normally incinerate the guishe, thus causing environmental contamination. Interestingly, recent studies showed that guishe contains molecules with nutritional properties, such as saponins, flavonoids and sugars. Therefore, in this work, we evaluated the effect of the crude extract of guishe as a feed additive in whiteleg shrimp diets. According to that, MS‐HPLC analysis of the extract showed the presence of saponins such as diosgenin, smilagenin, hecogenin, manogenin, tigogenin hexose, yucagenin, chlorogenin, diosgenin diglucoside and the flavonol, quercetin. After chemical analysis, the crude extract was included into an experimental diet in four levels; 0% (L0%), 0.1% (L0.1%), 0.3% (L0.3%) and 0.6% (L0.6%). Dietary incorporation of the extract was evaluated by zootechnical performance, haemolymph biochemistry, histomorphology and digestive enzyme activity of shrimps. After 5‐week feeding, the L0.3% diet showed significantly higher growth and better feed utilization among treatments. A significant increase in tubule epithelium height and tubule coverage area from hepatopancreas in shrimp under L0.3% diet compared with the control diet suggest an improvement of the health and nutritional status of the shrimp. Inclusion of L0.3% and L0.6% of the crude extract resulted in a reduction in amylase activity, without effect in glucose levels in the haemolymph. Thus, we suggest that lechuguilla guishe crude extract contains nutritional molecules that may be used as a feed additive to promote shrimp productivity.     

海参皂苷的研究现状

海洋中蕴涵着丰富的天然资源,海参作为一种食品和药材,具有很高的保健功能和药用价值。海参皂苷是海参体内的一种次生代谢产物,具有广泛的生理药理活性,因此潜藏了极大的应用价值。本文就近年来国内外关于海参皂苷性质的研究状况做一些简要的概述,为进一步研究海参皂苷提供较为详尽的参考资料。     

苜蓿皂苷对大鼠肝脏及肝脏细胞低密度脂蛋白受体、三磷酸腺苷结合盒转运体mRNA表达的影响

本试验旨在研究苜蓿皂苷对大鼠肝脏及大鼠肝脏细胞(BRL细胞)胆固醇清除和转运途径中关键基因低密度脂蛋白受体(LDLR)、三磷酸腺苷结合盒转运体G5(ABCG5)、三磷酸腺苷结合盒转运体G8(ABCG8)mRNA表达量的影响,从个体和细胞水平初步探讨苜蓿皂苷调控胆固醇清除和转运的分子机制。采用高脂饲粮建立大鼠高脂模型,测定苜蓿皂苷对正常、高脂大鼠血清指标[总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)含量]和肝脏LDLR、ABCG5、ABCG8 mRNA表达量的影响;采用高糖DMEM培养液建立BRL细胞脂变模型,测定苜蓿皂苷浓度对BRL细胞活性的影响,测定苜蓿皂苷对正常、脂变细胞LDLR、ABCG5、ABCG8 mRNA表达量的影响。结果表明:1)苜蓿皂苷显著降低高脂大鼠血清中TG、TC和LDL-C的含量(P0.05);2)苜蓿皂苷显著上调正常大鼠肝脏LDLR、ABCG5、ABCG8及高脂大鼠ABCG5、ABCG8 mRNA表达量(P0.05);3)添加200、250μg/m L苜蓿皂苷显著提高了BRL细胞的活性(P0.05);4)苜蓿皂苷显著上调正常BRL细胞LDLR、ABCG5、ABCG8 mRNA表达量(P0.05),而对脂变BRL细胞各基因mRNA表达量无显著影响(P0.05)。结果提示,苜蓿皂苷可通过调控LDLR、ABCG5、ABCG8 mRNA的表达来增加肝细胞内胆固醇的清除和转运,从而降低机体胆固醇的含量。     

刺玫果总皂苷的提取工艺研究

[目的]探索刺玫果中总皂苷的最优提取工艺条件。[方法]以刺玫果提取干浸膏中总皂苷的含量为指标,采用单因素和正交试验确定刺玫果中总皂苷的最优提取工艺条件。[结果]优化的刺玫果总皂苷的提取工艺条件为料液比1∶12,醇沉浓度60%,提取时间2.0h,提取次数3次。在此条件下提取的刺玫果干浸膏中总皂苷的含量平均达到58.16 mg/g。[结论]探索得到的工艺条件可用于刺玫果总皂苷的提取。     

有机溶剂盐溶液分离二醇型人参皂苷的研究

为确定分离二醇型人参皂苷最佳的有机溶剂及其用量。采用二醇型人参皂苷单体在有机溶剂CaCl2盐溶液中的不同溶解度进行萃取，经高效液相色谱检测，得出最优的有机溶剂及其用量。结果显示丙酮CaCl2盐溶液分离二醇型人参皂苷效果最佳。分离萃取二醇型人参皂苷最佳条件为皂苷(g):5% CaCl2(mL):丙酮(mL)=4:5:35。