Tanshinone ⅡA induces apoptosis via JNK signaling pathway in human osteosarcoma HOS cells

AIM: To explore the effect of tanshinone ⅡA on human osteosarcoma HOS cells and the underlying mechanism.METHODS: The cell viability and the appropriate dose of tanshinone ⅡA were determined by CCK-8 assay. Colony formation assay and Transwell assay were used to investigate the proliferation and migration abilities of the HOS cells treated with tanshinone ⅡA. The apoptosis of the HOS cells was monitored by Hoechst 33258 staining, transmission electron microscopy and flow cytometry. The protein levels of apoptosis-related molecules and JNK signaling-associated proteins were determined by Western blot. Meanwhile, a JNK inhibitor was added for confirming the relationship between the pathway and apoptosis mentioned above.RESULTS: Tanshinone ⅡA inhibited both HOS cell proliferation and migration in a dose-and time-dependent manner. Exposure of the HOS cells to tanshinone ⅡA resulted in the activation of apoptosis. Tanshinone ⅡA treatment increased the protein levels of cleaved caspase-3, Bax and JNK signaling-associated proteins, and decreased the protein level of Bcl-2, which were reversed by JNK inhibitor SP600125. Moreover, the result of CCK-8 assay revealed that tanshinone ⅡA-induced cell death was alleviated by JNK inhibitor.CONCLUSION: Tanshinone ⅡA induces cell growth inhibition and the activation of apoptosis via JNK signaling pathway in human osteosarcoma HOS cells.     

Effect of tanshinone ⅡA on liver lipid deposition and ferroptosis-related protein expression in ApoE-/- mice

AIM To observe the effect of tanshinone ⅡA on liver lipid deposition and ferroptosis-related protein expression in ApoE^-/- mice. METHODS Thirty-two ApoE^-/- mice were randomly divided into model group, high-dose (60 mg/kg) tanshinone ⅡA group, low-dose (30 mg/kg) tanshinone ⅡA group and simvastatin group, and C57BL/6J mice (n=8) were used as normal control group. The mice in normal control group were given the basic feeding, while the others were given high-fat diet. The mice in tanshinone ⅡA groups and simvastatin group were given corresponding drugs. The mice in normal control group and model group were intraperitoneally injected with equal volume of saline. Eight weeks later, the serum levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) were tested by automatic biochemistry analyzer. The liver tissues were stained with HE and oil red O. The contents of reactive oxygen species (ROS) and glutathione (GSH) in liver tissues of the mice were measured by commercially available kits. The liver glutathione peroxidase 4 (GPX4) and p53 were detected by immunohistochemical method. The protein and mRNA expression levels of ferroptosis-related factors GPX4, xCT/SLC7A11, p53 and ferritin heavy chain 1 (FTH1) were determined by Wes automatic Western blot quantitative analysis system and RT-qPCR. RESULTS Compared with normal control group, the serum levels of TC, TG and LDL-C in model group were increased significantly (P<0.05 or P<0.01), and HDL-C did not change significantly. The fat vacuoles were clearly visible in liver tissue. The content of ROS in liver tissue was increased significantly,and GSH was decreased significantly (P<0.01). The mRNA and protein expression levels of p53 were increased significantly, and GPX4, xCT/SLC7A11 and FTH1 were decreased significantly (P<0.05 or P<0.01). Compared with model group, tanshinone ⅡA significantly decreased the serum levels of TC, TG and LDL-C (P<0.05 or P<0.01), and HDL-C did not change significantly. High-dose and low-dose tanshinoneⅡA also significantly decreased the degree of steatosis, and the size of lipid droplets. The content of ROS in liver tissues was decreased significantly, and GSH was increased significantly (P<0.01). The mRNA and protein expression levels of GPX4, xCT/SLC7A11 and FTH1 were increased significantly, and p53 were decreased significantly (P<0.05 or P<0.01). CONCLUSION Tanshinone ⅡA reduces liver lipid deposition and lipid peroxidation damage in ApoE^-/- mice, which may be related to the intervention of ferroptosis-related proteins in the liver cells.     

降钙素基因相关肽对猪小肠上皮细胞钠离子依赖型Ⅱb磷转运蛋白表达及磷吸收的影响

本试验旨在研究不同浓度降钙素基因相关肽(CGRP)对猪小肠上皮细胞钠离子依赖型Ⅱb磷转运蛋白(NaPi-Ⅱb)表达和磷吸收的影响.选用第3～8代猪小肠上皮细胞进行试验,按单因素试验设计,将试验细胞分为1个对照组和5个CGRP(1×10-11～1×10-7 mmol/L)试验组,每组设6个重复.用4-甲基偶氮唑盐(MTT)法测定细胞生长速度,采用比色法测定CGRP干预24 h后细胞上清液磷含量,用RT-PCR法检测NaPi-Ⅱb mRNA相对表达量,以及蛋白质印迹(Western blot)法检测NaPi-Ⅱb蛋白表达.结果表明:与对照组相比,不同浓度CGRP对细胞生长没有显著影响(P＞0.05);各试验组均极显著或显著地促进了磷的吸收和NaPi-ⅡbmRNA的相对表达量(P＜0.01或P＜0.05);除1×10-10 mmol/L组外均显著提高了NaPi-Ⅱb蛋白的表达(P＜0.05).研究结果表明,CGRP通过诱导小肠上皮细胞中NaPi-Ⅱb的表达促进细胞磷的吸收,其中以1×10-9、1×10-8 mmol/L浓度的CGRP促进效果较为显著.     

高羊茅种质光合及叶绿素荧光参数对高温胁迫的响应

以21千高羊茅(Festuca arundinacea)种质为试材,在室内模拟35℃和45℃高温胁迫,测定了各高羊茅种质光合和叶绿素荧光参数,探讨高羊茅对高温胁迫的光合适应机制,并利用隶属函数法综合评价种质间耐热性强弱。研究结果表明,高温胁迫对光合参数和叶绿素荧光参数的影响非常显著(P0.05)。随着高温胁迫强度的增加,各高羊茅种质的净光合速率(P_n)、蒸腾速率(T_r)、最大光化学效率(F_v/F_m)、实际光量子产量(Yield)和光化学猝灭系数(gP)等参数均呈下降趋势;而叶片非光化学荧光猝灭系数(NPQ)则呈现先升高后降低的变化趋势。利用隶属函数法综合3个温度梯度下21个高羊茅种质的综合耐热D值,可以推测出各种质耐热性顺序为:Ⅲ-7Ⅲ-3Ⅱ-11I-17工-7工-4Ⅳ-3Ⅳ-2水城Ⅱ-13Ⅱ-10工-12Ⅳ-lⅣ-5Ⅱ-3黔草1号Ⅲ-8Ⅲ-11Ⅳ-4工-20Ⅱ-15。     

绵羊MHCⅡ类基因遗传多态性研究进展

主要组织相容性复合体(major histocompatibility complex,MHC)是广泛存在于脊椎动物体内的一类高度紧密连锁的基因群,绵羊MHC又称为绵羊淋巴细胞表面抗原(ovine lymphocyte antigen,OLA),位于绵羊20号染色体上,分为Ⅰ类、Ⅱ类和Ⅲ类区域,其中MHCⅡ类基因具有高度的基因多态性.文章综述了绵羊MHCⅡ类基因的分子结构及遗传多态性,重点总结了近十年来国内外不同绵羊品种MHCⅡ类基因遗传多态性的研究进展,并对绵羊MHCⅡ类基因未来的研究重点进行了展望.     

Protective effect of adiponectin on glucose and lipid metabolism by suppressing MHCⅡ expression

AIM: To investigate whether the protective effect of adiponectin on glucose and lipid metabolism is achieved through down-regulating major histocompatibility complex class Ⅱ (MHCⅡ) in the adipose tissue. METHODS: Adiponectin knockout (KO) mice and C57BL/6(WT) mice were fed with high-fat diet and standard diet for 24 weeks, respectively. The body weight, fasting blood glucose (FBG), fasting insulin (FINS), homeostasis model assessment of insulin resistance (HOMA-IR), triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), hepatic histology, and class Ⅱ trans-activator (CⅡTA), histocompatibility 2 class Ⅱ antigen E beta (H2-Eb1) and cluster of differentiation 74(CD74) mRNA and MHC Ⅱ protein levels in adipose tissue were measured at sacrifice. siRNA targeting MHC Ⅱ and overexpression vector was used in 3T3-L1 cells to explore the effect of adiponectin on the protein level of MHCⅡ. RESULTS: The levels of body weight, FBG, FINS, HOMA-IR, TC, TG, LDL-C, hepatic steatosis, CⅡTA, H2-Eb1 and CD74 mRNA expression, and MHCⅡ protein expression in the KO mice were higher than those in the WT mice that fed with high-fat diet or standard diet. In 3T3-L1 cells, inhibition of adiponectin reversed MHC Ⅱ protein level induced by specific siRNA. The expression of MHC Ⅱ in adipocytes decreased after adiponectin was overexpressed. CONCLUSION: Adiponectin improves glucose and lipid metabolism through suppressing the expression of MHCⅡ in the adipose tissue.     

农杆菌介导的柳枝稷遗传转化体系的优化

【目的】为建立农杆菌介导的柳枝稷高效再生及遗传转化体系。【方法】试验以柳枝稷品种Alamo、Performer及Blackwell的成熟种子为外植体诱导愈伤组织。愈伤诱导6周后,借助解剖镜观察愈伤形态,去除愈伤组织外层含水量大、海绵状愈伤组织,挑选位于愈伤组织中心位置的核状、紧实型愈伤置于继代培养基培养。暗培养3周后,在解剖镜下挑选结构松脆、生长旺盛的愈伤组织按细胞系继代增殖培养。该类愈伤组织易于分化,属单子叶植物愈伤组织分类中的第Ⅱ型。经两次继代增殖培养后,可以获得足够的愈伤组织进行再生和遗传转化研究。为优化柳枝稷Ⅱ型愈伤组织的农杆菌侵染流程,试验比较了3种农杆菌侵染流程下的转基因效率。真空和干燥处理(VD):将装有愈伤组织及菌液的50 m L离心管置于真空泵中抽真空10 min(压力-0.8 MPa),28℃,80r/min慢摇20 min后弃菌液,将愈伤组织堆放于3层滤纸上干燥处理2 h,随后将愈伤组织转入含有500μL无菌水的2层滤纸上进行共培养;冷处理加真空和干燥处理(CVD):侵染前将愈伤组织置于3%麦芽糖、300μmol·L-1谷氨酰胺(Gln)溶液中冰浴20 min,然后依VD流程侵染,共培养阶段用MP培养基代替无菌水;渗透冷处理加真空和干燥处理(PVD):用6%的麦芽糖溶液替代CVD中3%麦芽糖溶液。通过比较Gus染色及PCR阳性率来评价三种方法的转化效率。【结果】愈伤挑选方法不仅从柳枝稷低地型品种中挑选得到再生率达95%以上的Ⅱ型愈伤细胞系,也首次获得高地型品种Blackwell的Ⅱ型愈伤组织,分化率达50%。不同转化方法评价过程中发现,低地型品种AlamoⅡ型愈伤组织采用CVD农杆菌侵染流程转化效率最高,达72%,显著高于侵染流程VD(53%)和PVD(44%)。应用CVD转化法,Performer转化率达96%;首次成功进行了高地型品种Blackwell的遗传转化,转化效率为5.6%。【结论】本研究建立了借助解剖镜挑选柳枝稷易于再生的Ⅱ型愈伤组织的方法,该方法适用于柳枝稷不同生态型品种;农杆菌转化过程中采用CVD侵染流程可显著提高转基因效率。本研究首次报道了柳枝稷Ⅱ型愈伤组织的挑选流程,建立了适合不同生态型柳枝稷的高效再生及遗传转化体系,首次获得高地型品种Blackwell的转基因植株,为柳枝稷遗传改良及其功能基因研究奠定基础。该方法也适用与其他难于再生的单子叶植物再生及遗传转化体系的建立。     

密度和空间布局种植方式对夏玉米穗位叶光合生理性状的影响

为探明夏玉米适宜种植方式的光合生理机制,采用裂区试验设计研究了种植密度(9.3万,8.1万,6.9万,5.7万株/hm~2)、空间布局(等行距1穴1株,等行距1穴3株和宽窄行1穴3株)以及它们的交互作用对夏玉米郑单958开花后不同生育时期(开花期、抽丝期、灌浆前期、灌浆后期和完熟期)净光合速率及其相关性状的影响。结果表明:宽窄行1穴3株能显著降低夏玉米开花期和完熟期穗位叶净光合速率,而8.1万株/hm~2密度下的净光合速率不受空间布局的影响;等行距1穴1株空间布局下,种植密度不会显著影响穗位叶净光合速率。种植密度、空间布局以及它们交互作用对类胡萝卜素含量影响显著。等行距1穴3株空间布局下,9.3万株/hm~2种植密度显著降低前3个观测时期类胡萝卜素含量,而6.9万株/hm~2在宽窄行1穴3株以及5.7万株/hm~2在等行距1穴1株空间布局下显著降低完熟期类胡萝卜素含量。种植密度、空间布局及其交互作用均对PSⅡ最大光化学效率无显著影响。综上可知,8.1万株/hm~2种植密度不受空间布局的显著影响,而等行距1穴1株空间布局不受种植密度的显著影响,均能保证夏玉米郑单958植株的净光合作用及其相关指标维持在较高水平。结果也为从光合物质基础和光合水分生理基础方面(色素含量、荧光特性、净光合速率和蒸腾速率)解释种植方式-光合产物源-产量库之间的关系提供了理论数据。     

门冬胰岛素30注射液强化治疗Ⅱ型糖尿病的临床观察

目的观察门冬胰岛素30（诺和锐30）注射液强化治疗Ⅱ型糖尿病的效果.方法Ⅱ型糖尿病患者86例,给予生活方式干预,包括健康教育,低热量、低脂肪饮食,中等体力活动,戒烟、限酒,血压高者给予降压等,血脂异常者给予调脂.治疗组给予诺和锐30注射液3餐前皮注（根据血糖监测结果调整胰岛素剂量）,对照组给予诺和灵30R笔芯,早、晚餐前30 min皮注.观察6个月,观察治疗前后患者的FBG、2hPBG、HbA1c及低血糖发生率的变化.结果Ⅱ型糖尿病患者用诺和锐30强化治疗后,FBG、2hPBG、HbA1c等指标均好于对照组,低血糖发生率较对照组减少,P〈0.01,经比较有统计学意义.结论对Ⅱ型糖尿病患者应用诺和锐30强化治疗,血糖平稳,低血糖发生率减低,疗效肯定.     

东北梅花鹿线粒体COⅡ基因序列的遗传结构及其系统发育分析

采用基因测序方法,对3种东北梅花鹿的COⅡ基因片段进行了PCR扩增和测序。结果表明,3种东北梅花鹿COⅡ基因片段长度均为740bp,A＋T含量都大于G＋C含量。3个品种单倍型多样性较高（0.673～0.717）,就单个品种而言,敖东梅花鹿的遗传多样性最高。