海参加工废液中皂苷的提取及其抗肿瘤作用的研究

为了研究其对肿瘤小鼠的抑瘤作用,以及对免疫器官的影响。从海参废液中提取海参皂苷,通过建立S180皮下肿瘤小鼠模型,采用经口灌胃和腹腔注射2种方式给药后,将小鼠称重,断颈处死,然后剥离的肿瘤组织、胸腺、脾脏,称重并进行统计学分析。结果表明：海参皂苷能显著抑制S180皮下瘤小鼠的肿瘤生长;通过2种给药方式相比较,腹腔注射给药方式抑瘤效果(60.29%,P＜0.01)明显优于经口灌胃给药方式(37.70%;P＜0.05);与阳性对照环磷酰胺相比,海参皂苷对小鼠的生长影响较小,而且提高小鼠的脾脏指数和胸腺指数,对小鼠免疫器官有促进作用。本实验为今后海参皂苷抗肿瘤活性的进一步研究提供了有效依据。     

干柱层析法制备油茶总皂苷对照品初探

目的 研究干柱层析法制备油茶皂苷对照品的方法。方法 原料用60 %乙醇超声提取,浓缩至无醇味,石油醚脱酯后用正丁醇按3:1体积比加入萃取。取正丁醇层并真空浓缩至干,干法上样于100目~200目硅胶柱色谱分离,用乙酸乙酯-甲醇-正丁醇-水(2:1:4:7)的上相展层;根据Rf值取出相应的条带并用甲醇淋洗收集蒸干,薄层及理化性质检测。结果 该方法所制样品由薄层色谱为一点,纯度较高。结论 该方法简便、实用且制备量大,可作为油茶总皂苷对照品。     

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.     

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。     

三七及竹节参中皂甙的不同提取方法

采用冷浸、回流、微波等方法提取三七及竹节参皂甙,按光密度值分别计算出三七及竹节参皂甙的含量。结果显示,微波提取法与冷浸、回流法相比提取率稍高,并且微波提取法操作简单、安全、省时。因此,微波提取法将使三七和竹节参资源得到更好的开发和利用。     

水溶性盾叶薯蓣皂苷及其药理活性研究进展

盾叶薯蓣甾体皂苷包含水溶性皂苷和脂溶性皂苷2大类,其中水溶性盾叶薯蓣皂苷是传统中医药临床药理的重要活性基础,但没有进行系统性地研究。该文对近年来水溶性盾叶薯蓣皂苷的化学成分、分离纯化及药理活性等方面的研究进行综述,为甾体皂苷类新药研发及皂素工业生产中对水溶性盾叶薯蓣皂苷回收利用提供理论参考。     

双水相体系萃取油楠种子总皂苷的研究

利用双水相萃取技术提取分离油楠种子中的总皂苷,以乙醇/硫酸铵为双水相萃取体系,研究乙醇质量分数、硫酸铵质量分数、p H值、温度等因素对萃取效果的影响。结果表明:在乙醇质量分数为30%,硫酸铵质量分数为20%,p H6.0,温度为30℃的萃取条件下,油楠总皂苷在两相的分配系数为12.60,回收率为94.8%。该方法可用于油楠种子中总皂苷的提取分离。     

不同温度下雌激素对施氏鲟早期性腺发育及血清性激素的影响

在不同养殖温度下注射外源雌激素后对孵化后90日龄的施氏鲟性未分化的仔鱼进行培育,研究早期性腺发育过程中组织结构和血清性激素含量的变化,以了解温度和外源雌激素对施氏鲟幼鱼性分化和性别比率的影响,探讨施氏鲟的早期性别控制方法和机制。在不同试验温度下,注射外源雌激素-17β-estradiol(E2)30 d后,使施氏鲟幼鱼的卵巢分化提前,但未能改变施氏鲟雌雄1∶1的性别比率。注射24 h后各温度组血浆T含量下降到0 ng.mL-1,显著低于对照组0.46 ng.mL-1的水平(P<0.05)。注射30 d后,血浆T含量有所回升,但以高温组21、24和27℃组的血浆T含量开始回升,27℃组回升幅度最大,与处理前水平无显著差异(P>0.05)。注射后24 h,各温度组血浆E2含量显著升高(P<0.05),7 d后开始缓慢降低,15 d后降到最低值后开始回升,30 d时血浆E2含量与对照组差异不显著(P>0.05)。与注射后血清T含量一样,以温度高组回升快,最快组为21℃组。结果表明,注射17β-E2不能改变施氏鲟的性别比率,但对施氏鲟血浆T和E2含量具有一定影响作用。     

甾体皂苷的生物活性研究进展

概述了植物中甾体皂苷的生物活性,详细介绍了其在心血管系统、抗肿瘤、增强免疫、降血糖、抗生育、杀虫、抗真菌等方面的药理作用、构效关系、临床应用及研究进展,为开发和利用甾体皂苷药用植物资源奠定基础.     

Differentiation and development of Leydig cell, and induction of spermatogenesis during testicular differentiation in the Japanese eel, Anguilla japonica

The initial appearance and the development of Leydig cells (LCs), the sites of steroid hormone production in the testis, were investigated ultrastructurally during testicular differentiation in the Japanese eel, Anguilla japonica. In addition, the effects of a single injection of human chorionic gonadotropin (HCG; 5 IU g body weight^-1) on histological changes of the testes and serum 11-ketotestosterone (11-KT) were examined at various stages (15–18, 20–23, 26–29, 32–35, 38–41 and 46–50 cm body length (BL)) of testicular differentiation. Testicular differentiation was morphologically characterized by the development of loose connective tissue on the medial side in animals 18–29 cm in BL. Ultrastructurally, LCs were first identified in the loose connective tissue of the testis of the 23 cm fish. In the testes of fish over 32 cm, clusters of LCs were distributed throughout the interstitial region accompanying the increase in number of spermatogonia. In fish larger than 32 cm, spermatogenesis was induced by administration of HCG; serum 11-KT levels were also raised. On the other hand, there was no effect on spermatogenesis or serum 11-KT levels in fish less than 29 cm, or in the controls. These result suggests that morphological differentiation of LCs occurs in testis of the 23 cm eel, and subsequently, the testes of eels of BL more than 32 cm acquire the capability to produce steroid hormones.