Schisandrin B exerts anticancer effects on human gastric cancer cells through ROS-mediated MAPK,STAT3,and NF-κB pathways

Schisandrin B(Sch B)is a monomer with anti-cancer and anti-inflammatory effects,which are isolated from the plant Schisandra chinensis(Turcz)Baillon.We investigated the anti-gastric cancer(GC)effects of Sch B and its underlying molecular mechanisms.The Cell Counting Kit-8 assay was used to determine the effects of Sch B on the viability of GC and normal cell lines.Hoechst/propidium iodide staining and flow cytometry were used to assess the apoptosis induction of Sch B.Western blotting was used to evaluate the effects of Sch B on downstream apoptotic proteins.The DCFH-DA fluorescent probe was used to assess the regulatory effects of Sch B on reactive oxygen species(ROS)levels and related signaling pathways in GC cells.The results showed that Sch B could regulate the phosphorylation level of mitogen-activated protein kinase(MAPK)by upregulating ROS accumulation in gastric cancer cells,and then reduce the expression of nuclear factor kappa B(NF-κB)and phosphorylated transcription 3(p-STAT3).In addition,Sch B downregulated the cell cycle proteins cyclin-dependent kinase 2/4/6 and cyclin D1/E,and arrested cells in the G0/G1 phase.Moreover,it also inhibited cell migration,which was reversed with Nacetylcysteine pretreatment.In summary,Sch B has killing effects on GC cells by upregulating the production of intracellular ROS and regulating the MAPK/STAT3/NF-κB signaling pathway,leading to the migration arrest and apoptosis of GC cells.

Study on HPLC method to determine contents of Schisandrin A and Schisandrin B in Schisandra chinensis extraction

The determination method of Schisandrin A and Schisandrin B in Schisandra chinensis was improved with the high performance liquid chromagraphy （HPLC）. The sample was extracted exceedingly in the critical limit of CO2. The retention time of Schisandrin A and Schisandrin B was reduced, with methano/water （75 ： 25） as mobile phase. The wavelength for detection was 254 nm. The R^2 of standard curve was 0.9998 and the relative standard deviation was 2.31% and 3.17% with the recovery of 96.45% and 97.37%, respectively. The result shows that the rate of veracity of this method is higher and it proves that the determination method of Sehisandrin A and Schisandrin B in Schisandra chinensis is a feasible method.

Schisandrin B protects PC12 cells by decreasing the expression of amyloid precursor protein and vacuolar protein sorting 35

PC12 cell injury was induced using 20 μM amyloid β-protein 25-35 to establish a model of Alzheimer＇s disease. The cells were then treated with 5, 10, and 25 μM Schisandrin B. Methylthiazolyldiphenyl-tetrazolium bromide assays and Hoechst 33342 staining results showed that with increasing Schisandrin B concentration, the survival rate of PC12 cells injured by amyloid β-protein 25-35 gradually increased and the rate of apoptosis gradually decreased. Reverse transcription-PCR, immunocytochemical staining and western blot results showed that with increasing Schisandrin B concentration, the mRNA and protein expression of vacuolar protein sorting 35 and amyloid precursor protein were gradually decreased. Vacuolar protein sorting 35 and amyloid precursor protein showed a consistent trend for change. These findings suggest that 5, 10, and 25 μM Schisandrin B antagonizes the cellular injury induced by amyloid β-protein 25-35 in a dose-dependent manner. This may be caused by decreasing the expression of vacuolar protein sorting 35 and amyloid precursor protein.

Inhibitory effect of schisandrin B on gastric cancer cells in vitro

AIM: To investigate the inhibitory effect and possible mechanism of action of schisandrin B in SC-B on gastric cancer cells in vitro. METHODS: SC-B consisted of schisandrin B, aloe- emodin, and Astragalus polysaccharides. Exponentially growing human gastric cancer SGC-7901 cells were divided into six treatment groups: (1) control group (RPMI 1640 medium); (2) negative control group (2% DMSO); (3) positive control group (50 mg/L 5-Fluorouracil, 5-FU); (4) low-dose group (LSC, fi nal concentration of schisandrin B, 25 mg/L); (5) moderate-dose group (MSC, fi nal concentration of schisandrin B, 50 mg/L); (6) high- dose group (HSC, fi nal concentration of schisandrin B, 100 mg/L). Follow-up was done at 12-48 h. An MTT (Methylthiazolyldiphenyl-tetrazolium bromide) assay was used to examine the inhibitory effect of SC-B on gastric cancer cells. The mitosis index was assessed using an inverted microscope. Flow cytometry was used to visualize the cell cycle. An RT-PCR (Reverse transcription-Polymerase chain reaction) -based assay was used to detect mRNA expression for cyclin D1 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). RESULTS: The MTT assay showed that the number of living cells in the LSC, MSC and HSC groups was signif icantly smaller than that in the DMSO-treated group (P < 0.05) at 12- 48 h. The inhibitory rate (IR) of the LSC group was 41.15% ± 3.86%, 59.24% ± 5.34% and 69.93% ± 7.81% at 12, 24 and 48 h, respectively. The IR of the MSC group was 42.82% ± 4.94%, 62.68% ± 7.58% and 71.79% ± 8.12% at 12, 24 and 48 h, respectively. The IR of the HSC group was 37.50% ± 3.21%, 40.34% ± 2.98% and 61.99% ± 4.88% at 12, 24 and 48 h, respectively. These results suggested that a moderate dosage had the most obvious inhibitory efficacy at 48 h. Compared to the DMSO group, themitosis index of the LSC, MSC, HSC groups was greatly decreased (P < 0.05) at all time points. Any dose of SC-B suppressed mitosis within 12-48 h. Compared to the DMSO group, the percentage of cells in the G0/G1 phase of the MSC group was greatly increased, and that of the S + G2M phase was greatly decreased, while the percentage of cell inhibition (PCI) in the MSC group was greatly increased (P < 0.05). This suggested that SC-B could exclusively arrest cells in the G0/G1 phase. Cyclin D1 mRNA expression was lower in the MSC group than that in the DMSO group (P < 0.05). CONCLUSION: SC-B can inhibit the proliferation and aberrant mitosis of human gastric cancer SCG-7901 cells in vitro. This inhibitory effect may be due to the down- regulation of cyclin D1 mRNA expression, which causes cell cycle arrest of gastric cancer cells.

Inhibitory effect of schisandrin B on free fatty acid-induced steatosis in L-02 cells

AIM:To investigate the effects of schisandrin B (Sch B) on free fatty acid (FFA)-induced steatosis in L-02 cells.METHODS:Cellular steatosis was induced by incubating L-02 cells with a FFA mixture (oleate and palmitate at the ratio of 2:1) for 24 h.Cytotoxicity and apoptosis were evaluated by 3-(4,5-dmethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and Annexin V/propidium iodide staining,respectively.Cellular total lipid was determined using a photocolorimetric method after Nile red staining,and triglyceride content was measured using an enzymatic kit.To study the effects of Sch B on steatosis,L-02 cells were treated with Sch B (1-100 μmol/L) in the absence or presence of 1 mmol/L FFA for 24 h,and cellular total lipid and triglyceride levels were measured.To explore the mechanisms of action of Sch B in the steatotic L-02 cells,mRNA levels of several regulators of hepatic lipid metabolism including adipose differentiation related protein (ADRP),sterol regulatory element binding protein 1 (SREBP-1),peroxisome proliferator-activated receptor (PPAR)-α and PPAR-γ were measured by quantitative real-time polymerase chain reaction (PCR),and protein levels of ADRP and SREBP-1 were measured by immunoblotting.RESULTS:Treatment with 1 mmol/L FFA for 24 h induced intracellular lipid accumulation in L-02 cells comparable to that in human steatotic livers without causing apparent apoptosis and cytotoxicity.Sch B mitigated cellular total lipid and triglyceride accumulations in the steatotic L-02 cells in a dose-dependent manner.Quantitative real-time PCR and Western blot analyses revealed that treatment of L-02 cells with 100 μmol/L Sch B reverted the FFA-stimulated up-regulation of ADRP and SREBP-1.CONCLUSION:Sch B inhibits FFA-induced steatosis in L-02 cells by,at least in part,reversing the up-regulation of ADRP and SREBP-1.

Investigation of in Vitro and in Vivo Metabolism of Schisandrin B from Schisandrae Fructus by Liquid Chromatography Coupled Electrospray Ionization Tandem Mass Spectrometry

Schisandrin B (Sch B) is one of the active dibenzocyclooctadiene lignans found in the Schisandrae Fructus. Experimental studies have shown that Sch B possesses various pharmacological properties, including anti-cancer, neuroprotective and nephroprotective activities. However, no detailed information on its biotransformation was reported in the literature. Here, we investigated the in vitro and in vivo metabolism of Sch B by using ultra-performance liquid chromatography coupled with tandem mass spectrometry. In vitro study detected and identified one oxygenated metabolite. Four metabolites were detected and identified from the in vivo study. The results indicated that the metabolism of Sch B mainly involved the demethylation of methoxy groups, the opening of five-member ring and the glucuronidation of metabolites in rats. The metabolites were identified for the first time by MS/MS analyses.

Schisandrin B Protects against Ischemic Brain Damage by Regulating PI3K/AKT Signaling in Rats

Objective To explore the effect and mechanism of schisandrin B(Sch B)in the treatment of cerebral ischemia in rats.Methods The cerebral ischemia models were induced by middle cerebral artery occlusion(MCAO)and reperfusion.Sprague-Dawley rats were divided into 6 groups using a random number table,including sham,MCAO,MCAO+Sch B(50 mg/kg),MCAO+Sch B(100 mg/kg),MCAO+Sch B(100 mg/kg)+LY294002,and MCAO+Sch B(100 mg/kg)+wortmannin groups.The effects of Sch B on pathological indicators,including neurological deficit scores,cerebral infarct volume,and brain edema,were subsequently studied.Tissue apoptosis was identified by terminal transferase-mediated dUTP nick end-labeling(TUNEL)staining.The protein expressions involved in apoptosis,inflammation response and oxidative stress were examined by immunofluorescent staining,biochemical analysis and Western blot analysis,respectively.The effect of Sch B on phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)signaling was also explored.Results Sch B treatment decreased neurological deficit scores,cerebral water content,and infarct volume in MCAO rats(P<0.05 or P<0.01).Neuronal nuclei and TUNEL staining indicated that Sch B also reduced apoptosis in brain tissues,as well as the Bax/Bcl-2 ratio and caspase-3 expression(P<0.01).Sch B regulated the production of myeloperoxidase,malondialdehyde,nitric oxide and superoxide dismutase,as well as the release of cytokine interleukin(IL)-1βand IL-18,in MCAO rats(P<0.05 or P<0.01).Sch B promoted the phosphorylation of PI3K and AKT.Blocking the PI3K/AKT signaling pathway with LY294002 or wortmannin reduced the protective effect of Sch B against cerebral ischemia(P<0.05 or P<0.01).Conclusions Sch B reduced apoptosis,inflammatory response,and oxidative stress of MCAO rats by modulating the PI3K/AKT pathway.Sch B had a potential for treating cerebral ischemia.

五味子乙素通过TLR4/NF-κB信号通路对急性胰腺炎大鼠肺部损伤的影响

目的:探讨五味子乙素通过Toll样受体4(TLR4)/核转录因子-κB(NF-κB)信号通路对急性胰腺炎(AP)大鼠肺部损伤的影响。方法:取SD大鼠,通过胆胰管内逆行注射5%牛磺胆酸钠方法诱导建立AP肺损伤模型,经随机数表法分为模型组、五味子乙素组、TLR4过表达载体组、TLR4空载组、五味子乙素+TLR4过表达载体组,每组12只大鼠,再取12只SD大鼠仅翻动肠管不注射5%牛磺胆酸钠,作为假手术组。以药物分别干预大鼠后,检测各组大鼠肺功能及各组大鼠腹水量与肺组织湿重/干重(W/D);HE染色检测各组大鼠肺组织病理形态并评分;检测各组大鼠动脉血气;全自动生化分析仪检测大鼠血清淀粉酶,ELISA检测炎症细胞因子IL-6、IL-18水平;蛋白免疫印迹法检测肺组织TLR4/NF-κB通路蛋白表达;免疫组织化学染色检测肺组织TLR4蛋白表达。结果:与假手术组相比,模型组大鼠肺组织出现病理损伤改变,模型组大鼠MV、PEF、PaO_(2)、OI显著降低(P<0.05),Ri、腹水量与W/D、PaCO_(2)、Holfbauer评分、血清淀粉酶、IL-6与IL-18水平、肺组织TLR4阳性细胞比例、TLR4与MYD88蛋白表达、p-NF-κB p65/NF-κB p65水平显著升高(P<0.05)。与模型组、五味子乙素+TLR4过表达载体组分别相比,五味子乙素组大鼠肺组织病理损伤改变程度均减轻,MV、PEF、PaO_(2)、OI均升高(P<0.05),Ri、腹水量与W/D、PaCO_(2)、Holfbauer评分、血清淀粉酶、IL-6与IL-18水平、肺组织TLR4阳性细胞比例、TLR4与MYD88蛋白表达、p-NF-κB p65/NF-κB p65水平均降低(P<0.05);TLR4过表达载体组大鼠肺组织病理损伤改变程度均加重,MV、PEF、PaO_(2)、OI均降低(P<0.05),Ri、腹水量与W/D、PaCO_(2)、Holfbauer评分、血清淀粉酶、IL-6与IL-18水平、肺组织TLR4阳性细胞比例、TLR4与MYD88蛋白表达、p-NF-κB p65/NF-κB p65水平均升高(P<0.05)。与模型组相比,TLR4空载组大鼠各指标差异无统计学意义(P>0.05)。结论:五味子乙素可通过下调TLR4/NF-κB信号通路,抑制炎症,减轻AP大鼠肺部损伤,修复肺功能。

冰片修饰五味子乙素胶束处方优选及体外对bEnd.3细胞靶向性评价研究

目的优化冰片修饰五味子乙素胶束(Borneol-Schisandrin B-Micelles,Bor-Sch B-Ms)处方,并对其体外脑靶向作用进行初步探索。方法采用薄膜分散法制备Bor-Sch B-Ms;对Soluplus/TPGS1000质量比(mg/mg)、Soluplus/五味子乙素质量比(mg/mg)和水化温度(℃)进行优化,确定Bor-Sch B-Ms的最优处方制备工艺;并对最优处方工艺制得的胶束进行表征;采用高效液相色谱法(HPLC)对胶束中五味子乙素进行含量测定,以测得Bor-Sch B-Ms的包封率;利用不同药物对小鼠脑内血管内皮细胞(bEnd.3)的荧光摄取实验进行Bor-Sch B-Ms体外靶向性评价。结果最优处方为Soluplus 120 mg,Soluplus/TPGS1000=2∶1,Soluplus/五味子乙素=12∶1,DSPE-PEG20002 mg,水化温度40℃,处方量为5 mL。所得最优胶束的粒径为(93.72±0.65)nm,电位为(-2.73±0.35)mV,Bor-Sch B-Ms的包封率为(93.54±0.86)%。体外细胞摄取实验显示,与五味子乙素胶束组(Sch B-Ms)相比,Bor-Sch B-Ms组细胞的荧光强度明显增加(P<0.05)。结论通过Box-Behnken设计-响应面法确定Bor-Sch B-Ms最优处方,制备得到的Bor-Sch B-Ms具有潜在的脑靶向性。

五味子苷B调控miR-370-3p/CCL3轴对幼年肺炎小鼠免疫炎症因子水平的影响

目的:探讨五味子苷B(Sch B)在幼年肺炎小鼠的作用机制。方法:将90只幼年雄性小鼠随机分为对照组、模型组、Sch B低剂量组(Sch BL组)、Sch B高剂量组(Sch BH组)、Sch BH+antagomir-NC组、Sch BH+miR-370-3p antagomir组各15只。Sch BL组和Sch BH组小鼠分别灌胃20、60 mg/kg的Sch B;Sch BH+antagomir-NC组和Sch BH+miR-370-3p antagomir组,先将1 nmol的miR-370-3p antagomir、antagomir-NC用20μL的磷酸盐缓冲液(PBS)溶解,在Sch B灌胃后将miR-370-3p antagomir、antagomir-NC质粒分别经尾静脉注射到小鼠体内;对照组和模型组给予等量生理盐水。每天1次,持续给药7 d。测定各组小鼠肺组织的湿干质量比;采用苏木精-伊红(HE)染色观察各组小鼠肺组织的病理形态;检测各组小鼠肺组织中炎症因子水平;流式细胞术检测外周血T淋巴细胞亚群;实时定量反转录聚合酶链式反应(qRT-PCR)检测肺组织中miR-370-3p和CCL3的mRNA水平;双荧光素酶实验验证miR-370-3p与CCL3的靶向关系。结果:与对照组相比,模型组幼鼠肺组织结构紊乱,肺泡壁变厚,出现大量炎性细胞浸润,组织受损严重,肺组织湿干质量比、CD8^(+)、肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6、CCL3 mRNA水平均升高,CD4^(+)、CD4^(+)/CD8^(+)、miR-370-3p水平均降低(P均<0.05)。与模型组相比,Sch BL组和Sch BH组幼鼠肺组织中肺泡壁变薄,炎性细胞浸润明显减少,损伤减轻,肺组织湿干质量比、CD8^(+)、TNF-α、IL-6、CCL3 mRNA水平均降低,CD4^(+)、CD4^(+)/CD8^(+)、miR-370-3p水平均升高(P均<0.05)。加入miR-370-3p antagomir进行回补实验,结果显示Sch B对肺炎幼鼠免疫功能和炎症保护作用被逆转,且CCL3 mRNA水平升高(P<0.05);双荧光素酶报告基因实验验证了miR-370-3p与CCL3存在靶向关系。结论:Sch B能够通过靶向调节miR-370-3p/CCL3轴来增强幼年肺炎小鼠免疫功能,并抑制炎症反应。

Schisandrin B Inhibits NLRP3 Inflammasome Pathway and Attenuates Early Brain Injury in Rats of Subarachnoid Hemorrhage

Objective: To determine whether Schisandrin B(Sch B) attenuates early brain injury(EBI) in rats with subarachnoid hemorrhage(SAH). Methods: Sprague-Dawley rats were divided into sham(sham operation), SAH, SAH+vehicle, and SAH+Sch B groups using a random number table. Rats underwent SAH by endovascular perforation and received Sch B(100 mg/kg) or normal saline after 2 and 12 h of SAH. SAH grading, neurological scores, brain water content, Evan’s blue extravasation, and terminal transferase-mediated dUTP nick end-labeling(TUNEL) staining were carried out 24 h after SAH. Immunofluorescent staining was performed to detect the expressions of ionized calcium binding adapter molecule 1(Iba-1) and myeloperoxidase(MPO) in the rat brain, while the expressions of B-cell lymphoma 2(Bcl-2), Bax, Caspase-3, nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3(NLRP3), apoptosis-associated specklike protein containing the caspase-1 activator domain(ASC), Caspase-1, interleukin(IL)-1β, and IL-18 in the rat brains were detected by Western blot. Results: Compared with the SAH group, Sch B significantly improved the neurological function, reduced brain water content, Evan’s blue content, and apoptotic cells number in the brain of rats(P<0.05 or P<0.01). Moreover, Sch B decreased SAH-induced expressions of Iba-1 and MPO(P<0.01). SAH caused the elevated expressions of Bax, Caspase-3, NLRP3, ASC, Caspase-1, IL-1β, and IL-18 in the rat brain(P<0.01), all of which were inhibited by Sch B(P<0.01). In addition, Sch B increased the Bcl-2 expression(P<0.01). Conclusion: Sch B attenuated SAH-induced EBI, which might be associated with the inhibition of neuroinflammation, neuronal apoptosis, and the NLRP3 inflammatory signaling pathway.

五味子素A、B和五味子丙素的密度泛函研究

采用密度泛函B3LYP方法在6-31G基组水平上对五味子素A、B及五味子丙素3种五味子提取物进行了优化计算,并从平衡几何构型、前线分子轨道、净电荷分布等方面对计算结果做了比较.计算结果表明分子中的二氧五环对分子的药物活性具有较大影响.随着分子中二氧五环数目的增加,分子中联苯环扭转角减小,前线轨道能级和能级差都减小,联苯环上正电荷增加,由此可判断3种分子活性顺序应为五味子丙素>五味子素B>五味子素A.

基于脂质代谢组学评价五味子素B诱导的急性高甘油三酯血症小鼠模型

目的利用脂质代谢组学技术对五味子素B(schisandrin B,Sch B)诱导的小鼠高甘油三酯血症模型进行评价和提供新的实验依据。方法♂ICR小鼠分为4组:正常饮食(ND)组; ND+Sch B组;高脂高糖饮食(HFFD)组; HFFD+Sch B组。生化法检测血清甘油三酯(TG)和总胆固醇水平;利用超高效液相色谱-四极杆飞行时间质谱联用仪(UPLCQ-TOF/MS)的代谢组学技术方法测定各组小鼠血清中脂类代谢物的变化。结果 ND+Sch B组与ND组比,筛选出27个差异代谢物,分别为TG类18个、磷脂酰胆碱(PC) 7个、磷脂酰乙醇胺(PE) 2个; HFFD组与ND组比,筛选出27个差异代谢物,分别为神经鞘磷脂6个、PC 13个、胆甾醇酯(CE) 2个、TG类5个、磷脂酰肌醇1个; HFFD+Sch B组与HFFD组比,筛选出25个差异代谢物,分别为TG类14个、CE 1个、PC 6个、PE 4个。结论 Sch B诱导的高甘油三酯血症动物模型涉及血清脂质代谢组学的改变。

天然产物五味子乙素对口腔癌细胞增殖、凋亡及侵袭的影响

目的 研究天然产物五味子乙素(Schisandrin B,Sch B)对口腔癌细胞增殖、凋亡和侵袭的影响。方法 用不同浓度的五味子乙素分别处理口腔癌细胞株SCC15和CAL27细胞24 h和48 h,采用CCK-8法和AnnexinV-FITC/PI双标记流式细胞术,检测五味子乙素对口腔癌细胞增殖及凋亡的影响;采用Transwell实验分别检测五味子乙素对细胞侵袭的影响,采用qRT-PCR和Western Blot检测五味子乙素对Prx1 mRNA与蛋白表达的影响。结果 CCK-8和流式细胞术检测发现五味子乙素明显抑制SCC15和CAL27细胞增殖并促进细胞凋亡,呈时间和剂量依赖性(P<0.05)。Transwell实验结果显示,五味子乙素对侵袭的抑制作用呈时间和剂量依赖性。五味子乙素对Prx1 mRNA的表达无显著影响,但对其蛋白表达有显著的抑制作用(P<0.05)。结论 五味子乙素可能通过调控Prx1蛋白的表达抑制口腔癌细胞的增殖、侵袭,促进细胞凋亡。

五味子乙素对LPS诱导小神经胶质BV-2细胞损伤保护的机制研究

目的研究五味子乙素对脂多糖(LPS)诱导的小神经胶质BV-2细胞炎症损伤的保护效应及其作用机制。方法体外常规培养BV-2细胞,用LPS诱导细胞炎症损伤模型,同时将细胞分为正常对照组(对照组)、LPS诱导模型组(模型组)、LPS+10μmol/L五味子乙素组、LPS+20μmol/L五味子乙素组及LPS+40μmol/L五味子乙素组,采用CCK-8试剂盒测定细胞的增殖情况,ELISA试剂盒检测细胞上清中炎症因子IL-6及TNF-α的释放量,Western blot分析炎症相关蛋白及Traf6/TAK1信号通路的活化情况。结果五味子乙素可以明显改善LPS对细胞增殖的抑制作用及LPS诱导细胞的炎症反应,主要表现在提高细胞的相对存活率、降低炎症因子IL-6及TNF-α的水平及抑制炎症相关蛋白iNOS及COX-2的表达,与模型组比较差异有统计学意义(P<0.05)。Western blot结果显示,五味子乙素可以抑制Traf6及p-TAK1蛋白的表达水平,与模型组比较差异有统计学意义(P<0.05)。结论五味子乙素能明显改善LPS诱导BV-2细胞的炎症损伤,其机制可能与下调iNOS、COX-2蛋白的表达及降低Traf6/TAK1信号通路活化相关。

五味子乙素对脂多糖诱导的脓毒症急性肺损伤大鼠肺组织病理损伤、炎症反应和核因子-κB表达的影响

目的观察五味子乙素对脂多糖(lipopolysaccharide,LPS)诱导的脓毒症急性肺损伤大鼠肺组织病理损伤、炎症反应和核因子-κB(nuclear factor kappa-B,NF-κB)表达的影响。方法采用气管滴注40 mg/kg LPS制备大鼠急性肺损伤大鼠模型,手术前1 h腹腔注射80 mg/kg五味子乙素进行干预治疗,测定大鼠肺湿/干质量比,BCA试剂盒测定大鼠支气管肺泡灌洗液(bronchoalceolar lavage fluid,BALF)总蛋白,牛鲍氏计数板计算BALF中白细胞数,苏木精-伊红染色观察大鼠肺组织损伤情况,ELISA法检测大鼠肺组织中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-6(interleukin-6,IL-6)含量,相关试剂盒检测大鼠肺组织中丙二醛(malondialdehyde,MDA)和超氧化物歧化酶(superoxide dismutase,SOD)含量,蛋白质印迹法检测大鼠肺组织中NF-κB p65、磷酸核因子-κB抑制蛋白(phosphorylation inhibitor of nuclear factor kappa-B,pIκBα)和Toll样受体4(Toll-like receptor 4,TLR4)的表达水平。结果与模型组相比,五味子乙素组大鼠肺湿/干质量比、BALF中总蛋白含量及白细胞数均显著减少(P<0.05),病理损伤显著减轻,TNF-α、IL-1β、IL-6含量均显著减少(P<0.05),MDA含量显著下降,SOD活性显著升高(P<0.05),NF-κB p65、pIκBα和TLR4表达水平均显著下调(P<0.05)。结论五味子乙素能够减轻LPS诱导的脓毒症急性肺损伤大鼠肺组织的病理损伤,抑制炎症反应和氧化应激反应,抑制NF-κB通路激活。

五味子乙素调控NF-κB通路对顺铂致HK-2细胞炎症损伤的保护作用

目的研究五味子乙素对顺铂致HK-2细胞炎症损伤的保护作用及相关机制。方法以不同浓度的五味子乙素及顺铂干预人肾小管上皮HK-2细胞,CCK-8法检测细胞活性,Hoechst 33258荧光染色法观察细胞形态变化,Western blot法检测NF-κB活化,酶联免疫吸附试验(ELISA)法检测细胞上清液中炎性细胞因子[肿瘤坏死因子-α(TNF-α)、白细胞介素1β(IL-1β)、白细胞介素6(IL-6)]水平,实时荧光聚合酶链反应(RT-PCR)法检测细胞中TNF-α,IL-1β,IL-6的m RNA表达。结果与对照组相比,顺铂组细胞活性显著下降,细胞形态明显改变;五味子乙素呈浓度依赖性地减轻顺铂的细胞毒性作用。同时,五味子乙素能显著抑制顺铂所致NF-κB活化及炎性细胞因子(TNF-α,IL-1β,IL-6)在细胞上清液中及m RNA表达的升高。结论五味子乙素能通过调控NF-κB通路对抗顺铂所致肾小管上皮细胞的炎症损伤。

五味子乙素抗梅花鹿源BVDV的实验研究

采用中性红染料法,测定了五味子乙素对MDBK细胞的最大安全浓度,探讨了五味子乙素抗梅花鹿源BVDV病毒的作用。结果表明:五味子乙素对MDBK细胞无损伤的最大安全浓度是19.53μg/mL;五味子乙素具有显著抗梅花鹿源BVDV作用,且有一定的量效关系,在安全浓度范围内,随浓度的提高,五味子乙素抗病毒的作用增强。

五味子乙素介导Traf6/NF-κB信号通路抑制心肌细胞肥大实验研究

目的:研究五味子乙素抑制脂多糖(lipopolysaccharide,LPS)诱导心肌细胞肥大的作用及其机制。方法:原代培养心肌细胞,用LPS(1 mg/L)诱导心肌细胞肥大,考察IκBα抑制剂BAY11-7082和不同剂量五味子乙素对心肌细胞肥大的影响。采用计算机图像分析系统观察细胞大小;考马斯亮蓝法分析细胞总蛋白量;ELISA试剂盒检测白细胞介素6(interleukin-6,IL-6)和肿瘤坏死因子α(tumor necrosis factorα,TNF-α)表达量;免疫印迹技术检测心房钠尿肽(atrial natriuretic peptide,ANP)和Traf6/NF-κB信号通路蛋白表达水平。结果:与LPS诱导组相比,五味子乙素和BAY11-7082均可以抑制LPS诱导的心肌细胞肥大,体现在蛋白含量降低,细胞体积减小,ANP蛋白表达降低(P<0.05);五味子乙素还能够降低炎症反应,表现在细胞外液中炎症因子IL-6和TNF-α水平降低(P<0.05),Traf6、p65蛋白表达量降低(P<0.05)及IκBα蛋白水平升高(P<0.05),且呈浓度依赖性。此外,高剂量五味子乙素与BAY11-7082对心肌细胞肥大的保护作用相近。结论:五味子乙素能抑制LPS诱导原代心肌细胞肥大,对心肌细胞的保护作用主要通过抑制Traf6/NF-κB信号通路的活化来实现。

基于基因芯片技术的五味子素B诱导高甘油三酯血症小鼠模型作用机制研究

目的探讨五味子素B(Schisandrin B,Sch B)诱导高脂血症小鼠模型的基因(肝脏)表达谱的变化及可能的机制。方法ICR小鼠,雄性,分为7组,每组10只:正常饮食(normal diet,ND)组(2组)、ND/Sch B(0.5,2 g·kg^(-1))组、高脂/糖饮食(high fat/fructose diet,HFFD)组、HFFD/Sch B(0.5,2 g·kg^(-1))组。小鼠用Sch B(橄榄油配制)灌胃,48 h后用生化法检测血清和肝脏甘油三酯(TG)和总胆固醇(TC)水平,血清高密度脂蛋白胆固醇(HDL)和低密度脂蛋白胆固醇(LDL)水平;ELISA法检测血清极低密度脂蛋白胆固醇(VLDL)、载脂蛋白A1(Apo A1)、Apo E、Apo B48、Apo B100、Apo CⅡ和Apo CⅢ水平;HE染色观察肝脏病理学变化;油红O染色观察肝脏脂质沉积;通过基因芯片技术分析各组小鼠肝脏中全基因表达谱的变化及所涉及的通路。结果ND/Sch B组小鼠血清/肝脏TG(升高266%/352.57%)和TC水平显著升高(升高36.54%/25.70%),血清HDL和LDL水平显著升高(分别升高29.96%和30.68%),但VLDL、Apo E、Apo B100、Apo CⅡ和Apo CⅢ显著降低(分别降低15.98%、28.90%、20.76%、20.53%和17.82%);肝脏脂质沉积并出现病理学损伤;与HFFD组比,HFFD/Sch B组血清TG水平、肝脏TG和TC水平升高,血清HDL、LDL、VLDL、Apo E、Apo B100和Apo CⅡ水平显著降低;ND/Sch B组与ND组比肝脏有1016个基因差异表达,其中上调基因722个,下调基因294个,HFFD/Sch B组与HFFD组比肝脏有1162个基因差异表达,其中上调基因671个,下调基因491个。而HFFD组与ND组比肝脏有2070个基因差异表达,其中上调基因1289个,下调基因781个。Pathway分析结果显示Sch B诱导的高脂血症与11条通路的改变有关。结论Sch B诱导高脂血症的机制与肝脏调节脂代谢多条通路、载脂蛋白及脂蛋白代谢密切相关。