Dioscin from Polygonatum sibiricum induces apoptosis and autophagy in Ishikawa human endometrial cancer cell and in vivo

With an aim to comprehend the precise regulatory mechanism of dioscin against endometrial carcinoma(EC), we firstly extracted the components from Polygonatum sibiricum followed by identification and structural characterization. The anti-EC activity of dioscin was initially determined based on the inhibition of Ishikawa cell proliferation and tumor growth. The high-throughput sequencing data indicated that dioscin not only promoted apoptosis, including decrease of poly ADP-ribose polymerase(PARP) and B-cell lymphoma-2(Bcl-2) and increase of c-PARP and Bcl-2-associcated agonist of cell death(Bad), but also induced autophagy, including increase of autophagic lysosomes and LC3Ⅱ/LC3Ⅰ ratio. Mechanistic exploration suggested that dioscin induced autophagy and apoptosis through inhibition of PI3K/AKT/mTOR signaling pathway. Besides, the dioscin-regulated p53 pathway was mainly involved in autophagy induction. Furthermore, inhibition of Ishikawa cell autophagy was linked to dioscin-induced apoptosis. Our data suggest the immense potential of dioscin for the development of functional food for EC and related medical application.

Impact and mechanism study of dioscin on biological characteristics of colorectal cancer cells

BACKGROUND Colorectal cancer(CRC)is a considerable global health issue.Dioscin,a compound found in several medicinal plants,has shown potential anticancer effects.AIM To find the relationship between CRC cells(HCT116)and diosgenin and clarified their mechanisms of action.METHODS CRC cell line HCT116 was cultured by dividing cells into control and dioscin groups(dioscin+Jagged 1 group;Jagged 1 group,5μg/mL;and dioscin group,2.5μg/mL).The dioscin groups were given different concentrations of dioscin.Cell Counting Kit-8 was chosen for testing cell viability in different groups.Flow cytometry was established to undiscover the apoptosis rate of human liver cancer cell line 11.Real-time PCR as well as Western blot analyses were applied to reveal the expression levels of caspase-3,Notch,and other proteins.Transwell and scratch experiments were conducted to assess cell migration and invasion abilities.RESULTS This study indicated that dioscin restricted the growth of HCT116 cells,boosted cell apoptosis,and rose the Bax/Bcl-2 ratio as well as the expression of Caspase-3.Dioscin also inhibited physiological activities,for instance cell migration,and significantly reduced the expression levels of proteins for instance Notch1(P<0.05).Dioscin partially reversed the effects of Jagged 1.CONCLUSION Dioscin exerts a certain inhibitory effect on HCT116,and its mechanism of action may be linked,with the inhibition of the Notch1 signaling pathway.

Dioscin-induced Apoptosis of Human LNCaP Prostate Carcinoma Cells through Activation of Caspase-3 and Modulation of Bcl-2 Protein Family

Dioscin is a natural steroid saponin derived from several plants, showing potent anti-cancer effect against a variety of tumor cell lines. In the present study, we investigated the anti-cancer activity of dioscin against human LNCaP cells, and evaluated the possible mechanism involved in its antineoplastic action. It was found that dioscin（1, 2 and 4 μmol/L） could significantly inhibit the viability of LNCaP cells in a time- and concentration-dependent manner. Flow cytometry revealed that the apoptosis rate was increased after treatment of LNCaP cells with dioscin for 24 h, indicating that apoptosis was an important mechanism by which dioscin inhibited cancer. Western blotting was employed to detect the expression of caspase-3, Bcl-2 and Bax in LNCaP cells. The expression of cleaved caspase-3 was significantly increased, and meanwhile procaspase-3 was markedly decreased. The expression of anti-apoptotic protein Bcl-2 was down-regulated, whereas the pro-apoptotic protein Bax was up-regulated. Moreover, the Bcl-2/Bax ratio was drastically decreased. These results suggested that dioscin possessed potential anti-tumor activity in human LNCaP cells through the apoptosis pathway, which might be associated with caspase-3 and Bcl-2 protein family.

Protective effects of dioscin against Parkinson's disease via regulating bile acid metabolism through remodeling gut microbiome/GLP-1 signaling

It is necessary to explore potent therapeutic agents via regulating gut microbiota and metabolism to combat Parkinson's disease(PD).Dioscin,a bioactive steroidal saponin,shows various activities.However,its effects and mechanisms against PD are limited.In this study,dioscin dramatically alleviated neuroinflammation and oxidative stress,and restored the disorders of mice induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP).16 S rDNA sequencing assay demonstrated that dioscin reversed MPTP-induced gut dysbiosis to decrease Firmicutes-to-Bacteroidetes ratio and the abundances of Enterococcus,Streptococcus,Bacteroides and Lactobacillus genera,which further inhibited bile salt hydrolase(BSH)activity and blocked bile acid(BA)deconjugation.Fecal microbiome transplantation test showed that the anti-PD effect of dioscin was gut microbiota-dependent.In addition,non-targeted fecal metabolomics assays revealed many differential metabolites in adjusting steroid biosynthesis and primary bile acid biosynthesis.Moreover,targeted bile acid metabolomics assay indicated that dioscin increased the levels of ursodeoxycholic acid,tauroursodeoxycholic acid,taurodeoxycholic acid and bmuricholic acid in feces and serum.In addition,ursodeoxycholic acid administration markedly improved the protective effects of dioscin against PD in mice.Mechanistic test indicated that dioscin significantly up-regulated the levels of takeda G protein-coupled receptor 5(TGR5),glucagon-like peptide-1 receptor(GLP-1R),GLP-1,superoxide dismutase(SOD),and down-regulated NADPH oxidases 2(NOX2)and nuclear factor-kappaB(NF-kB)levels.Our data indicated that dioscin ameliorated PD phenotype by restoring gut dysbiosis and regulating bile acid-mediated oxidative stress and neuroinflammation via targeting GLP-1 signal in MPTP-induced PD mice,suggesting that the compound should be considered as a prebiotic agent to treat PD in the future.

Protective effects of Dioscin on TNF-α-induced collagen-induced arthritis rat fibroblast-like synoviocytes involves in regulating the LTB4/BLT pathway

Background and Objective:LTB4 has been shown to be involved in rheumatoid arthritis(RA)pathogenesis.The effect of Dioscin(Dio)on the LTB4 pathway of RA have not been reported yet.This study aimed at further exploring whether Dioscin’s effects on TNF-αinduced collagen-induced arthritis(CIA)rat fibroblast-like synoviocytes(FLS)connected with the LTB4 and its receptor pathway.Materials&Methods:In this experiment,control group,TNF-αgroup,and different concentrations of Dioscin groups were established.Cell viability was evaluated using MTT assay.The levels of LTB4 in the samples of above groups were measured using ELISA.The mRNA expression levels of LTA4H,BLT1,and BLT2 were detected by quantitative real time PCR,while the expression level of LTA4H proteins were detected using western blot.The distribution of LTA4H was assessed by immunofluorescence assay.Results:the LTB4 level of TNF-αgroup in sample supernatant was higher than both control group and Dioscin groups with decreased LTB4 levels(p<0.05).Compared with the control group,the expression of LTA4H was significantly increased in TNF-αgroup(p<0.05),whereas LTA4H expressions were significantly decreased in all Dioscin groups when compared to TNF-αgroup(p<0.05).The mRNA expressions of BLT1 and BLT2 were markedly higher in TNF-αgroup than those in control group while Dioscin treatment significantly inhibited the increased expressions of BLT1 and BLT2 induced by TNF-α(p<0.05).Conclusions:These results firstly demonstrate that the protective effect of Dioscin on TNF-αinduced FLS may involve in its reducing LTB4 production by down-regulating LTA4H expression,and may inhibit its downstream pathway by decreasing LTB4 receptors levels.This findings suggest that dioscin produces a potential therapeutic effects for RA via its influencing LTA4H/LTB4/BLT pathway.

Neuroprotective effect of dioscin on cerebral ischemia/reperfusion mice by reducing glial cell activation and maintaining the levels of SODs

In the present study, we aimed to explore the neuroprotective effect of dioscin, a natural steroid saponin, on transient focal cerebral ischemia induced by middle cerebral artery occlusion （MCAO） in mice and its related mechanism, We observed that dioscin （1,5 mg/kg, intracerebroventricular injection 30 min before MCAO） dramatically reduced the cerebral infarct volume, leading to improved neurological symptoms and reduced death of neuron, astrocytes and microglia in the infarct region. The gliosis and the reduced expressions of SOD1 and SOD2 by MCAO in the hippocampal CA1 region were significantly elevated by 1.5 mg/kg dioscin administration. These findings suggested that pretreatment of dioscin had a neuroprotective effect on mice transient focal cerebral ischemia via inhibiting the gliosis and elevating the SOD levels.

Effects of Dioscin Extracted from Polygonatum Zanlanscianense Pamp on Several Human Tumor Cell Lines

Dioscin was extracted and isolated from Polygonatum Zanlanscianense Pamp. The effects of dioscin on HL60, HeLa, H14, and MDA MB 435 cell lines were studied with the results showing that dioscin dramatically inhibited the growth of the MDA MB 435, H14, HL60, and HeLa cell lines. The IC 50 of dioscin on these cell lines were 2.6, 0.8, 7.5, and 4.5 μmol/L respectively.

薯蓣皂苷抑制Wnt/β-catenin信号通路促进肝癌细胞凋亡的研究

目的研究薯蓣皂苷(DIO)对人HepG2肝癌细胞凋亡的影响及其可能抗癌作用机制。方法DIO(0.25、0.5、1、2、4、6和8μmol/L)干预HepG2肝癌细胞,MTT法检测细胞增殖情况,并计算半数抑制率(IC50);划痕实验分析细胞的迁移能力,Western blot检查细胞中凋亡因子及Wnt/β-catenin通路蛋白表达差异。结果与对照组比较,DIO组HepG2细胞抑制率显著升高(P<0.01),并诱导细胞在形态学上出现凋亡特点,呈时间与剂量依赖性;划痕实验结果显示DIO显著抑制HepG2细胞的迁移距离;与对照组比较,DIO干预细胞24 h后,Caspase-3、cleaved Caspase-3水平明显上调,而Bcl-2、β-catenin水平显著下调(P<0.05、P<0.01);采用LiCl激活Wnt/β-catenin信号通路,LiCl组细胞中Wnt1、β-catenin水平较对照组显著上调(P<0.01);与LiCl组比较,LiCl+DIO组HepG2细胞Wnt1、β-catenin、GSK-3β蛋白表达显著下调(P<0.01)。结论DIO体外能够促进HepG2肝癌细胞凋亡,机制可能是其下调β-catenin的蛋白表达,抑制Wnt/β-catenin信号通路传导,进而抑制凋亡因子Bcl-2表达,最终诱导细胞凋亡而发挥抗肝癌作用。

薯蓣皂苷对蛋鸡脂肪肝综合征肝损伤和氧化应激的缓解作用及其机制研究

旨在探究薯蓣皂苷(dioscin, Dio)对高能低蛋白饲喂诱导蛋鸡脂肪肝综合征脂代谢紊乱和氧化损伤的缓解作用及其机制的影响。以105只150日龄的蛋鸡作为试验对象,分为5组:对照组,HELP(高能低蛋白饲料)组,低剂量组(添加100 mg/kg Dio),中剂量组(添加500 mg/kg Dio),高剂量组(添加1 000 mg/kg Dio),饲喂3个月后剖检观察,测定肝组织AMP-依赖性蛋白激酶(AMPK)和核因子红细胞系2相关因子2(Nrf2)转录水平。体外以棕榈酸钠(PA)诱导鸡肝癌(LMH)细胞代谢紊乱,CCK-8和吖啶橙/碘化丙啶检测细胞死亡,尼罗红染色观察脂滴,荧光探针法检测活性氧(ROS)产生水平,Western blot检测脂代谢、抗氧化相关蛋白表达水平,免疫荧光检测Nrf2入核。结果:与对照组相比,HELP组蛋鸡肝脏发生明显的脂肪变性和出血,中剂量组明显缓解肝脏脂肪变性和出血,提高AMPKα和Nrf2转录水平,Dio缓解PA诱导的LMH细胞死亡,减少LMH细胞中脂滴沉积,并显著上调AMPK通路(P<0.05),同时减少ROS产生并增加Nrf2的表达及入核。综上,Dio能够缓解高能低蛋白饲喂诱导蛋鸡脂肪肝综合征脂代谢紊乱和氧化损伤,激活LMH细胞AMPK和Nrf2信号通路缓解PA诱导的LMH细胞脂代谢紊乱及氧化应激的发生,为其作为一种生理调节剂用于家禽脂肪肝及其相关的营养代谢症的防控提供理论依据。

薯蓣皂苷对脓毒症大鼠肾损伤的影响及机制

目的 探究薯蓣皂苷对脓毒症大鼠肾损伤的影响及可能机制。方法 采用盲肠结扎和穿刺法建立脓毒症大鼠模型。将建模成功的60只大鼠按照随机数字表法分为模型组(0.5%羧甲基纤维素钠溶液),薯蓣皂苷低、中、高剂量组(30、60、120 mg/kg)和地塞米松组(阳性对照,10 mg/kg),每组12只;另取12只大鼠作为假手术组(0.5%羧甲基纤维素钠溶液)。造模15 min后,各组大鼠尾静脉注射给药/0.5%羧甲基纤维素钠溶液。给药24 h后,检测大鼠血清中肌酐(Cr)、尿素氮(BUN)、中性粒细胞明胶酶相关脂质运载蛋白(NGAL)、肾损伤分子1(KIM-1)、白细胞介素6(IL-6)、IL-1β、肿瘤坏死因子α(TNF-α)水平以及肾组织中丙二醛(MDA)水平、超氧化物歧化酶(SOD)活性和核因子E2相关因子2(Nrf2)、血红素加氧酶1(HO-1)、NOD样受体蛋白3(NLRP3)表达量,并观察肾组织病理形态学变化。结果 与模型组比较,薯蓣皂苷低、中、高剂量组大鼠肾组织病理损伤明显改善,血清中Cr、BUN、NGAL、KIM-1、IL-6、IL-1β、TNF-α水平和肾组织中MDA水平、NLRP3蛋白表达量均显著降低(P<0.05),肾组织中SOD活性和Nrf2、HO-1蛋白表达量均显著升高(P<0.05),且薯蓣皂苷的作用具有剂量依赖性(P<0.05);薯蓣皂苷高剂量组和地塞米松组大鼠肾组织病理损伤情况相似,上述指标水平差异均无统计学意义(P>0.05)。结论 薯蓣皂苷可能是通过激活Nrf2/HO-1信号通路抑制NLRP3炎性小体,实现对炎症因子表达和氧化应激的抑制,从而发挥对脓毒症肾损伤的保护作用。

薯蓣皂苷通过GSK3β/Nrf2/HO-1通路改善尿酸诱导的HK-2细胞氧化应激损伤的作用及机制研究

目的探讨薯蓣皂苷(dioscin)对尿酸(uric acid,UA)诱导的人肾小管上皮细胞(HK-2)氧化应激损伤的影响及分子机制。方法将HK-2细胞分为正常组、模型组(尿酸刺激造模)、条件对照组(尿酸+DMSO)和薯蓣皂苷组(尿酸+薯蓣皂苷)。通过尿酸诱导HK-2细胞氧化应激损伤模型;采用CCK-8法检测细胞活力,流式细胞技术检测细胞活性氧(ROS)水平,Real-time PCR法检测糖原合成激酶3(GSK3β)、核转录因子红系2相关因子2(Nrf2)和血红素加氧酶1(HO-1)在mRNA水平的表达,Western Blot法检测GSK3β、磷酸化糖原合成激酶3(p-GSK3β)、Nrf2及HO-1在蛋白水平的表达。结果经尿酸刺激后,HK-2细胞的活力明显下降,ROS水平明显升高(均P<0.001)。经薯蓣皂苷治疗后,HK-2细胞的活力增加,ROS水平明显降低(均P<0.001)。在蛋白及mRNA水平上,尿酸刺激后Nrf2和HO-1的表达均下降,薯蓣皂苷干预后Nrf2和HO-1表达均明显增加(均P<0.001)。在蛋白水平上,模型组细胞p-GSK3β/GSK3β比值较正常组明显下降,经薯蓣皂苷干预后p-GSK3β/GSK3β比值升高(均P<0.001)。结论薯蓣皂苷可能是通过促进GSK3β的磷酸化,激活Nrf2/HO-1通路,从而缓解尿酸诱导的HK-2细胞氧化应激损伤。

薯蓣皂苷的药理作用及机制研究进展

薯蓣在中国有着上千年的应用历史,祖国医学认为其归脾、肺、肾三经,可“平补三焦气阴”,兼具补涩之功,有补中益气、强身健骨,治诸虚百损、延年益寿等功效,被誉为“天然小人参”,也是最早药食同源的植物之一。其中穿龙薯蓣更是具有祛风除湿、舒筋通络、活血止痛、止咳平喘等作用。薯蓣因其性平和,功效全面,而被中医广泛应用于各种疾病的治疗之中。近年来,随着科学技术的不断发展,其主要活性成分薯蓣皂苷及水解产物薯蓣皂苷元被广泛发掘并应用于心脑血管疾病、关节炎、癌症、糖尿病等多种疾病的防治当中。现代研究表明,薯蓣皂苷具有改善心脑血管、抗肿瘤、降糖、降脂、保肝、抗炎止痛、提高免疫等多种药理作用,就近几年薯蓣皂苷的药理作用及其机制研究进展进行综述,旨在为该天然中药单体的进一步研发和应用提供参考。

薯蓣皂苷经PI3K/AKT通路对肝癌Bel-7402细胞增殖和凋亡的影响

目的分析薯蓣皂苷对肝癌Bel-7402细胞增殖和凋亡的影响并探讨其机制。方法肝癌Bel-7402细胞分为空白组和薯蓣皂苷低、中、高剂量组(给予1、2、8μmol/L的薯蓣皂苷)及薯蓣皂苷+抑制剂组(给予8μmol/L的薯蓣皂苷+10μmol/L的磷脂酰肌醇3激酶/蛋白激酶B(PI3K/AKT)信号通路抑制剂LY294002),于处理后12、24、36、48及72 h时采用四甲基偶氮唑盐(MTT)比色法测定细胞活力,于24 h时采用流式细胞术检测细胞凋亡情况、采用蛋白免疫印迹法(Western blot)检测p-PI3K和p-AKT的表达。结果与空白组相比较,薯蓣皂苷低、中、高剂量组细胞活力及p-PI3K、p-AKT表达均下降,凋亡率升高(P<0.05),且各剂量组间两两比较,上述指标水平差异均有统计学意义(P<0.05);与薯蓣皂苷高剂量组比较,薯蓣皂苷+抑制剂组细胞活力及p-PI3K、p-AKT表达下降,凋亡率升高(P<0.05)。结论薯蓣皂苷可能通过抑制PI3K/AKT通路抑制肝癌Bel-7402细胞增殖,诱导Bel-7402细胞凋亡。

薯蓣皂苷抗氧化应激机制研究进展

氧化应激是指机体受到外界刺激后,产生大量的活性氧自由基导致细胞氧化损伤的一种机制,与心血管疾病、神经系统疾病、炎症性疾病、肿瘤等多种疾病的发生和发展密切相关。寻找有效的抗氧化剂成为疾病预防和治疗的重要途径之一。薯蓣皂苷是一种天然存在的化合物,具有抗氧化和抗炎作用。近年来,研究发现薯蓣皂苷在抗氧化应激中起到重要的作用。本综述概述了氧化应激的概念和机制,并讨论了其可能的作用机制,薯蓣皂苷可通过清除自由基和过氧化物,增加机体内源性抗氧化剂的产生和释放,抑制Nrf2-ARE通路、NF-KB通路、MAPK通路、Keap1-Nrf2通路、AMPK通路等氧化应激信号通路的激活发挥作用。本研究总结了薯蓣皂苷在抗氧化应激方面的研究进展,以期为后续研究提供参考。

薯蓣皂苷治疗骨关节炎的研究进展

薯蓣皂苷是一种植物来源甾体皂苷,具有抗癌、免疫调节、心血管保护等多种药理作用,早期研究已经证实,薯蓣皂苷可以用于骨关节炎(OA)的治疗。本文通过整理相关文献,对薯蓣皂苷治疗OA的相关机制进行总结和分析,包括对软骨细胞、成骨细胞、破骨细胞、软骨基质代谢调节,抑制炎症反应的研究,以期为薯蓣皂苷在OA方面的相关研究提供理论参考。

薯蓣皂苷增强DNA损伤促进口腔鳞癌细胞凋亡

目的:研究薯蓣皂苷对人口腔鳞癌细胞SCC4、SCC25 DNA损伤和凋亡的影响及其可能的机制。方法:SCC4、SCC25细胞用不同浓度薯蓣皂苷(0.0、0.1、0.3、1.0、3.0、10.0和30.0μmol/L)处理48 h,通过CCK-8实验检测细胞活力;不同浓度(0.0、1.0、2.0、4.0μmol/L)薯蓣皂苷处理SCC4细胞48 h,采用免疫荧光技术检测γH2AX在SCC4细胞核内的表达情况;通过Western blotting检测DNA损伤修复蛋白的表达情况;采用流式细胞术检测细胞凋亡率,最后通过Western blotting检测细胞凋亡相关蛋白的表达变化。结果:薯蓣皂苷浓度依赖性地抑制了SCC4和SCC25细胞的活性(P<0.05);与薯蓣皂苷0.0μmol/L组比较,各薯蓣皂苷处理组SCC4细胞核中γH2AX的荧光表达强度显著上调(P<0.05);与薯蓣皂苷0.0μmol/L组比较,各薯蓣皂苷处理组SCC4、SCC25细胞DNA损伤信号蛋白P-ATM、P-CHK1、P-CHK2、γH2AX表达上升;与薯蓣皂苷0.0μmol/L组比较,各薯蓣皂苷处理组SCC4、SCC25细胞Cleaved-PARP、Cleaved-Caspase-3蛋白表达均升高。结论:薯蓣皂苷能抑制口腔鳞癌细胞的活性,并促进细胞的凋亡,其作用机制可能和其促进细胞的DNA损伤有关。

薯蓣皂苷对大鼠糖尿病肾病肾小管上皮细胞的影响

目的:探讨薯蓣皂苷对糖尿病肾病大鼠肾小管上皮细胞的影响及机制。方法:建立糖尿病肾病大鼠模型,分成对照组、模型组和薯蓣皂苷低、中、高剂量组,薯蓣皂苷低、中、高剂量组给予不同浓度(20、40、80 mg/kg)薯蓣皂苷灌胃,对照组与模型组给予等量生理盐水。收集正常大鼠肾小管上皮细胞培养,分为空白对照组、高糖组和薯蓣皂苷低、中、高剂量组,薯蓣皂苷低、中、高剂量组给予不同浓度(20、40、80μmol/L)薯蓣皂苷处理。H-E染色观察肾组织病理变化,RT-qPCR检测血小板衍化生长因子-BB(PDGF-BB)mRNA水平,ELISA法检测24 h尿白蛋白水平及肾小管上皮细胞中肿瘤坏死因子-α(TNF-α)、白细胞介素1β(IL-1β)、活性氧(ROS)、丙二醛(MDA)水平,流式细胞术检测细胞凋亡率,免疫印迹检测PDGF-BB蛋白表达。结果:与模型组比较,薯蓣皂苷组尿白蛋白及肾组织PDGF-BB mRNA水平降低,呈剂量依赖,肾组织病理损伤好转;细胞实验中,与高糖组比较,薯蓣皂苷组肾小管上皮细胞中PDGF-BB mRNA及蛋白水平以及TNF-α、IL-1β、MDA、ROS含量和细胞凋亡率降低,呈剂量依赖性。结论:薯蓣皂苷能抑制糖尿病肾病大鼠肾小管上皮细胞炎症反应、过氧化损伤及细胞凋亡,降低PDGF-BB表达,保护肾小管上皮细胞。

响应面法优化泡沫分离葫芦巴中薯蓣皂苷工艺

通过单因素试验及响应曲面试验设计对泡沫法分离葫芦巴中薯蓣皂苷工艺进行优化。探讨pH、稀释倍数(浓度)、温度、气速对泡沫法分离葫芦巴中薯蓣皂苷回收率及富集比的影响。结果表明,稀释倍数为12、操作温度23℃、pH值为8.5、空气流速为60 mL/min,葫芦巴中薯蓣皂苷的最优回收率为35.86%,富集比4.67,总皂苷质量分数为67.86%,该工艺条件稳定可行,为葫芦中薯蓣皂苷提取分离的工业化生产提供一定的理论指导和参考依据。

薯蓣皂苷通过调控AMPK/mTOR自噬信号通路影响骨肉瘤细胞增殖及侵袭的机制研究

目的探讨薯蓣皂苷(Dioscin)通过调控腺苷酸活化蛋白激酶(AMPK)/哺乳动物雷帕霉素靶蛋白(mTOR)自噬信号通路对人骨肉瘤MG63细胞增殖及侵袭的影响。方法取对数期生长人骨肉瘤MG63细胞分为空白对照组、Dioscin不同剂量组(20、40、80、100μmol/L)。CCK-8法检测细胞增殖能力,划痕实验检测细胞迁移能力,细胞迁移实验检测细胞侵袭能力,丹酰尸胺法观察细胞自噬情况,蛋白质印迹法检测磷酸化腺苷酸活化蛋白激酶(p-AMPK)/AMPK、磷酸化哺乳动物雷帕霉素靶蛋白(p-mTOR)/mTOR、转录因子EB(TFEB)、人微管相关蛋白轻链3Ⅱ(LC3-Ⅱ)、自噬效应蛋白beclin-1表达水平。结果与空白对照组比较,Dioscin20、40、80、100μmol/L组MG63细胞增殖率[(65.21±4.09)%、(51.94±3.95)%、(46.57±3.72)%、(37.31±4.23)%比100.00%]、划痕愈合率[(85.43±3.84)%、(70.62±3.17)%、(55.83±2.92)%、(44.23±3.27)%比(98.25±1.23)%]、侵袭细胞比率[(72.43±2.83)%、(63.76±3.34)%、(51.92±3.13)%、(42.44±3.09)%比(91.14±2.91)%]依次降低,呈浓度依赖性(P<0.05);空白对照组中仅有少量细胞自噬泡,作用48 h后Dioscin各组自噬泡依次增多;与空白对照组比较,Dioscin20、40、80、100μmol/L组MG63细胞中p-AMPK/AMPK[(0.35±0.04)、(0.50±0.02)、(0.60±0.03)、(0.88±0.03)比(0.20±0.02)]、TFEB[(0.42±0.02)、(0.68±0.03)、(0.92±0.04)、(1.36±0.10)比(0.30±0.02)]、LC3-Ⅱ[(0.41±0.03)、(0.70±0.04)、(0.88±0.03)、(1.17±0.07)比(0.26±0.02)]、beclin-1[(0.32±0.03)、(0.50±0.04)、(0.68±0.05)、(0.75±0.06)比(0.16±0.02)]蛋白表达依次增加,p-mTOR/mTOR[(0.75±0.02)、(0.52±0.03)、(0.40±0.04)、(0.31±0.02)比(0.86±0.05)]蛋白表达依次降低,差异均具有统计学意义(P<0.05);与Dioscin(100μmol/L)组比较,Dioscin(100μmol/L)+AMPK抑制剂(Compound C 10μmol/L)组p-AMPK/AMPK[(0.49±0.03)比(0.63±0.04)]、TFEB[(0.16±0.03)比(0.24±0.02)]、LC3-Ⅱ[(0.35±0.02)比(0.44±0.04)]、beclin-1[(0.44±0.03)比(0.56±0.04)]蛋白表达水平明显降低,p-mTOR/mTOR[(0.63±0.05)比(0.45±0.04)]表达明显升高(P<0.05)。结论Dioscin可能通过调控AMPK/mTOR-TFEB通路抑制骨肉瘤细胞增殖、迁移及侵袭。

与科研融合的药剂学实验教学设计——薯蓣皂苷PLGA纳米粒的制备及表征

将前沿纳米技术运用于药物制剂,设计了采用纳米沉淀法制备薯蓣皂苷PLGA纳米粒的药剂学实验课程。利用动态光散射法结合透射电镜对PLGA纳米粒的理化性质进行表征,采用超滤离心法测定薯蓣皂苷在PLGA纳米粒中的载药量和包封率。本实验可以锻炼本科生掌握运用前沿纳米技术制备新型制剂的能力,熟悉对新型纳米制剂的表征方法,进一步使学生了解新型纳米技术在药物制剂中的应用,提升学生运用新型纳米制剂技术的实践能力和科研素养。