Use of crude extract of garlic (Allium sativum L.) in reducing cytotoxic effects of arsenic in mouse bone marrow

A relatively high concentration of crude extract of garlic, (Allium sativum L.)--single clove variety, was found to reduce the cytotoxic effects of three doses of sodium arsenite, corresponding to 1/10, 1/30 and 1/50 of the LD₅₀ in laboratory bred male Swiss albino mice. The animals were given a single oral dose of the chemical, together with the extract, and effects were observed after 24 h in bone marrow cells following the colchicine–fixation–airdry–Giemsa schedule. The endpoints scored were chromosomal aberrations and damaged cells.     

Inhibition of clastogenic effect of cyclophosphamide and mitomycin C by Neem leaf-extract in mice

Azadirachta indica commonly known as ‘Neem’ is well known for its medicinal properties in the indigenous Indian system of medicine. Almost every part of the tree has some beneficial use. The anticlastogenic activity of ‘Neem’ against cyclophosphamide (CP) and mitomycin C (MMC) was studied in vivo in bone marrow cells of mice. Aqueous leaf-extracts of A. indica were injected intraperitoneally at doses 3, 6, 12 and 24 mg/kg body weight. Simultaneously, two known clastogens CP (10 mg/kg) and MMC (1.5 mg/kg) were administered individually to animals treated with 6 and 12 mg/kg of the leaf-extract. The end-points screened were chromosomal aberrations and damaged (aberrant) cells. Neem leaf-extract per se was found to be a weak clastogen; 6 and 12 mg/kg of the leaf-extract inhibited the clastogenicity of CP and MMC. The extent of inhibition was different for the two clastogens. An ANOVA test showed that the reduction in the frequency of chromosomal aberrations was significantly less when the leaf-extract was given in combination with CP. MMC co-administered with the leaf-extract showed a trend that was not statistically significant. The difference may be attributed to the degree of modulation of bioactivation of cytochrome P-450 enzymes, or the repair of damaged DNA or a difference in detoxification of the reactive species of the two genotoxicants. © 1998 John Wiley & Sons, Ltd.     

熟地和制首乌多糖对贫血小鼠骨髓有核细胞数和细胞周期的影响

目的:观察熟地多糖(RPS)和制首乌多糖(PPS)对骨髓抑制贫血小鼠外周血象、骨髓有核细胞(BMC)数和细胞周期的影响。方法:用60Co辐射和腹腔注射环磷酰胺(Cy)及氯霉素(Ch),建立骨髓抑制贫血小鼠模型,分别提取熟地和制首乌多糖,腹腔注射RPS(20mg/kg)和PPS(25mg/kg),观察对外周血象、BMC数的影响,并用流式细胞仪分析细胞周期。结果:RPS和PPS均能一定程度促进外周血象的恢复,明显提高骨髓抑制贫血小鼠BMC数量(P<0.01);流式细胞术分析表明,G0/G1期细胞比例明显减少(RPS:P<0.01,PPS:P<0.05),S期细胞比例明显增加(RPS:P<0.01,PPS:P<0.01)。结论:RPS、PPS能促进骨髓抑制贫血小鼠骨髓抑制的恢复,促进骨髓有核细胞进入增值周期。     

Comparison of the effects of crude extract of spinach-beet leaves and equivalent amounts of chlorophyll and chlorophyllin in modifying the clastogenic activity of chromium (VI) oxide in mice in vivo

The anticlastogenic activities of a crude extract of leaves of spinach-beet (Beta vulgaris var. benghalensis Hort.) and equivalent amounts of chlorophyll extracted from the leaves and of synthetic chlorophyllin in reducing cytotoxicity were compared following exposure of mice in vivo to a known clastogen chromium (VI) oxide. Male Swiss albino mice were administered orally the vegetable extract for 7 consecutive days and then exposed to the clastogen by gavage (20 mg/kg b wt). For comparison, equivalent amounts of extracted chlorophyll and synthetic chlorophyllin were administered to the mice, 2 h before exposure to the same dose of the metal. Chromosomes were studied from bone marrow cells 24 h after exposure, following colchicine-hypotonic-fixative-flame drying-Giemsa staining schedule. Chlorophyllin and the crude extract, when given alone, did not induce chromosomal aberrations and reduced the clastogenic effects induced by chromium (VI) oxide to a statistically significant level, indicating a protective action. Chlorophyll, however, produced a significant increase of chromosomal aberrations compared with control, when administered alone and was not able to reduce the clastogenicity of the metallic salt to a significant level.     

Inhibitory effect of chlorophyllin on the frequency of micronuclei induced by sodium nitrite in mice

In this report the potency of chlorophyllin (CHL) was evaluated to prevent two types of damage produced by nitrite in mice: the increase of micronucleated polychromatic erythrocytes (MNPE) and the bone marrow toxicity, measured as the index of polychromatic erythrocytes/normochromatic erythrocytes (PE/NE). The study was done in eight groups of male mice. The first three groups were administered orally for 4 days with sodium nitrite (10, 15 and 20 mg/kg), the daily administration with nitrite was followed by an intraperitoneal administration of CHL (4 mg/kg), three more groups were administered with the same amounts of nitrite, a seventh group of mice was treated with distilled water while another was treated with CHL (4 mg/kg). Our study produced two main results: (a) no bone marrow injury was induced by any of the tested chemicals, as indicated with the PE/NE index, and (b) CHL protected (as high as 44%) the MNPE produced in nitrite treated mice.     

四物汤对环磷酰胺致血虚证小鼠骨髓蛋白质组的影响

目的：研究四物汤对环磷酰胺（CTX）损伤致血虚证小鼠骨髓蛋白质组的影响,旨在分子水平上探讨四物汤促进骨髓造血的作用机制。方法：采用CTX 250 mg·kg^-1腹腔注射制成化学损伤型血虚证小鼠模型,利用蛋白质组学技术研究四物汤治疗后的差异蛋白质组。结果：研究发现CTX使小鼠骨髓中12个蛋白表达上调、3个蛋白表达下调,四物汤可使CTX造模后这些表达改变的蛋白有不同程度恢复。结论：四物汤可能通过影响与细胞凋亡、造血干祖细胞增殖分化有关的蛋白质而促进骨髓造血。     

未分化骨髓基质干细胞体外抑制淋巴细胞增殖反应的初步研究

目的:研究犬未分化骨髓基质干细胞(bone marrow stromal cells,BMSCs)体外抑制淋巴细胞增殖的作用。方法:从犬骨髓组织中分离获取骨髓基质干细胞,体外培养至第二代,1×105细胞/孔BMSCs刺激异体淋巴细胞,观察淋巴细胞增殖情况。以植物血凝素(PHA,2ug/mL)刺激淋巴细胞后,分别以1×102～5细胞/孔数量的异体BMSCs加入反应体系中,观察淋巴细胞增殖抑制情况。分别以1×102～5细胞/孔数量的"第三者"BMSCs加入双向混合淋巴细胞反应体系中,观察淋巴细胞增殖抑制情况。通透性隔离腔培养的BMSCs加入双向混合淋巴细胞反应体系中,观察淋巴细胞增殖抑制情况。结果:BMSCs刺激同种异体淋巴细胞体外增殖作用不明显;BM-SCs可明显抑制PHA刺激的淋巴细胞增殖(P<0.05);"第三者"BMSCs可明显抑制双向混合淋巴细胞反应(P<0.05),抑制作用呈BMSCs细胞数量依赖性;隔离腔培养的BMSCs也可明显抑制双向混合淋巴细胞反应(P<0.05)。结论:犬未分化BMSCs在体外具有抑制淋巴细胞增殖的作用,具有一定的调节淋巴细胞反应的能力。     

大鼠骨髓基质细胞移植治疗急性脊髓损伤的实验研究

目的:本实验在大鼠脊髓损伤后经原位移植骨髓基质细胞(bone marrow stroma cell-s,BMSCs),证明植入的BMSCs在损伤的脊髓内能分化为神经元样细胞,并通过对远期运动能力测试,探讨移植后神经功能障碍恢复的情况。方法:①采用Alien重物坠落法制作改良型大鼠Alien’s脊髓损伤模型(重量10g,高度30mm),苏木精-伊红(Haematoxyli-n/eosin,HE)染色鉴定脊髓损伤情况。②采用体外骨髓贴壁法培养大鼠BMSCs,进行细胞染色观察细胞形态,检测CD44从而对骨髓基质细胞进行鉴定。细胞进行三次培养传代后用携带绿色荧光蛋白(Green Fluorescent Protein,GFP)基因的慢病毒转染。③传代细胞移植及移植后细胞存活:将GFP标记的BMSCs原位(损伤区)植入损伤模型,分别对2w、4w、6w后处死大鼠行脊髓组织切片,免疫荧光显微镜下观察表达GFP的细胞。④随机入组的各组大鼠行改良Rivlin斜板实验和脊髓运动功能BBB评分法,比较单纯损伤组、移植后2w组、移植后4w组及移植后6w组大鼠的运动能力的改变。结果:①采用Alien’s重物坠落法造模,制造改良型大鼠Alien’s脊髓损伤模型,效果可靠、稳定。②BMSCs经过3次传代后增殖能力已趋于稳定,细胞在体外培养、扩增后,细胞形态改变为胞体呈锥形,突起交织成网,故可作为脊髓损伤后细胞移植的来源;染色观察细胞形态,检测CD44呈阳性。③由携带GFP基因的慢病毒转染BMSCs移植入脊髓损伤模型后,在6w内可见到细胞表达GFP。④通过改良Rivlin斜板实验和脊髓运动功能BBB评分法对单纯损伤组大鼠测试结果均明显差于移植对照组,差异具有统计学意义(P0.05);移植后2w、4w及6w组均能改善远期运动功能,以6w后神经功能障碍恢复最好,差异具有统计学意义(P0.05)。结论:BMSCs移植能促进SCI后神经功能障碍的恢复,其机制可能与BMSCs分化为神经元和神经胶质细胞及移植诱导后的BMSCs可与周围的神经细胞相互作用,产生某些细胞因子有关。     

阿魏酸钠诱导骨髓间充质干细胞向神经细胞分化的初步研究

目的 探讨阿魏酸钠诱导大鼠骨髓间充质干细胞向神经细胞分化的可能作用。方法 分离培养大鼠骨髓间充质干细胞，采用阿魏酸钠对其进行诱导培养，倒置显微镜下观察诱导培养不同时间后细胞的形态变化，并应用免疫细胞化学方法鉴定诱导培养后细胞表面神经细胞特异性标志物神经丝蛋白(NF)及神经元特异性烯醇化酶(NSE)的表达。结果 阿魏酸钠诱导培养6h后即可见细胞形态发生明显变化，24h后表现为典型的神经细胞样形态，NF和NSE神经细胞特异性标志物呈阳性表达。结论 初步证实阿魏酸钠具有诱导体外培养的大鼠骨髓间充质干细胞向神经细胞分化的作用。     

蜜环菌多糖对环磷酰胺损伤小鼠骨髓细胞的保护作用

目的:观察蜜环菌多糖对环磷酰胺所致小鼠骨髓细胞损伤的保护作用。方法:纯系昆明小鼠 50 只,随机分为5组,正常对照组;阳性对照组(rhG-CSF 20μg·kg^-1·d^-1);环磷酰胺(150 mg·kg^-1·d^-1)损伤组;蜜环菌多糖(25 0 mg·kg^-1·d^-1)低剂量保护组;蜜环菌多糖(50.0 mg·kg^-1·d^-1)高剂量保护组。阳性对照组sc rhG CSF 6 d,ip环磷酰胺3 d。保护组 ip蜜环菌多糖8 d,ip环磷酰胺3 d。记数外周血WBC,RBC,PLT和骨髓细胞悬液WBC,BM NC,骨髓象分析。结果: 保护组、阳性对照组外周血中的WBC,RBC,PLT,骨髓细胞悬液中WBC,BMNC明显高于损伤组,P<0.01;保护组中骨髓早幼粒、分叶核细胞数,高剂量组比低剂量组明显增多。结论: 蜜环菌多糖对环磷酰胺所致小鼠骨髓细胞损伤有较好的保护作用。     

骨髓间充质干细胞成骨分化的研究进展

骨髓间充质干细胞的成骨分化是近10年来骨组织工程种子细胞的研究热点,本文查阅相关文献,对近年骨髓间充质干细胞成骨分化进行综述.     

补虚化瘀方对化疗荷瘤小鼠骨髓造血微环境的影响

目的观察补虚化瘀方对联合荷瘤小鼠骨髓造血微环境的影响。方法采用P388瘤株制作荷瘤小鼠模型后,再以环磷酰胺、阿糖胞苷腹腔注射获得联合化疗荷瘤小鼠骨髓抑制模型,实验分为正常组、模型组及补虚化瘀方组,进行小鼠骨髓成纤维细胞集落形成单位(CFU-F)、骨髓VCAM-1及NF-κB、骨髓微血管测定。结果联合化疗可致荷瘤小鼠CFU-F计数、骨髓VCAM-1及NF-κB表达、骨髓微血管损伤明显低于正常组(P均<0.01);补虚化瘀方可提高联合化疗荷瘤小鼠CFU-F计数、骨髓VCAM-1及NF-κB表达,改善模型小鼠骨髓微血管损伤。结论补虚化瘀方具有保护联合化疗荷瘤小鼠骨髓抑制模型骨髓造血微环境和促进其化疗损害修复作用。     

丹酚酸B在大鼠骨髓间充质干细胞分化过程对心肌早期基因Nkx2.5 GATA-4 mRNA表达的影响

目的:探讨丹参单体丹酚酸B(Salvianolic acid B,SalB)在体外诱导大鼠骨髓间充质干细胞(bone marrowmes-enchymal stem cells,MSCs)分化为心肌样细胞的情况及Nkx2.5、GATA-4 mRNA的表达,从而确定大鼠MSCs体外诱导为心肌样细胞的条件、规律及转化细胞的特点。方法:选用成年近交系Wistar大鼠,利用percoll分离液进行密度梯度离心法和直接贴壁法进行分离、提纯MSCs,并进行培养扩增。免疫组化方法对第9代MSCs表面抗原进行鉴定。分别应用10μmol/L5-氮胞苷(5-azacytidine,5-aza)及250μg/L SalB联合5-aza(5-aza+SalB)对第9代的MSCs进行联合诱导24h,于诱导后第4周,用实时荧光定量RT-PCR法检测GATA-4和Nkx2.5基因表达。结果:①第9代MSCs免疫组化染色CD44呈阳性表达,CD34呈阴性表达。②诱导4周后,细胞体积变小,可见由几个细胞连接形成的多核肌管样结构。5-aza组、5-aza+SalB组均出现明确的心肌早期分化基因与5-aza组相比,5-aza+SalB组的NKx2.5、GA-TA-4 mRNA表达明显增加。结论:SalB具有促进5-aza诱导的MSCs向心肌样细胞分化的作用,并且在Nkx2.5基因表达上丹酚酸B的优势更加明显。     

Inhibition by chlorophyllin of forward and reverse bacterial mutagenicity of benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide

Chlorophyllin (CHL), a water soluble derivative of the green pigment chlorophyll, was evaluated for its antimutagenic activity towards benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide (BPDE). CHL suppressed dose-dependently the formation of 8-azaguanine-resistant mutant colonies in Salmonella typhimurium TM677 treated with BPDE. Under the same experimental conditions, the cytotoxicity of this direct-acting mutagen was concomitantly blunted by CHL. The protective effect of CHL against BPDE mutagenicity was confirmed in the Salmonella histidine⁺ reversion assay CHL also reduced the in vitro covalent binding of BPDE to DNA. Copyright © 1998 John Wiley & Sons, Ltd.     

大鼠内皮祖细胞的分离、培养与定向诱导分化

目的探讨建立一种从大鼠骨髓中分离培养与定向诱导分化内皮祖细胞(EPCs)的稳定、高效方法。方法密度梯度离心法从大鼠骨髓中分离单个核细胞,经差速贴壁结合特殊培养基扩增并向内皮细胞定向诱导分化EPCs。应用免疫荧光和流式细胞技术鉴定内皮细胞系列标志:CD34、CD31、Flk-1和祖细胞标志CD133。并通过检测其对FITC标记的UEA-1的吸附和内吞DiI-ac-LDL来进行细胞功能学的鉴定。对分化细胞行vWF、CD31免疫组化染色鉴定,并与血管内皮细胞合成前列腺素能力进行比较。结果梯度密度离心和贴壁法选择的细胞表达内皮细胞特异性抗原CD34、CD31、Flk-1,部分表达CD133。分离所得细胞经EBM-2专用培养基培养后,第5天可见集落形成,培养第9天CD133阳染细胞减少,CD31阳染细胞增加,细胞可特异性吸附FITC标记的荆豆凝集素并内吞DiI-ac-LDL,约3周左右形成铺路石样内皮细胞特有形态。诱导后的内皮祖细胞的前列腺素合成能力与血管内皮细胞之间无统计学差异(P>0.05)。结论该方法可以同时培养扩增和定向诱导分化内皮祖细胞,效率高,稳定性和重复性好。     

维肝力对骨髓抑制小鼠骨髓及血清FL分泌水平的影响

目的：研究维肝力对骨髓抑制贫血小鼠骨髓及血清Flt 3配体(FL)分泌水平的影响，探讨维肝力在骨髓造血调控方面的作用。方法：制作骨髓抑制贫血小鼠模型，小鼠随机分为正常组、骨髓抑制贫血模型组、维肝力组(高、中、低剂量100，50，25 g·L^-1)；观察维肝力对骨髓抑制贫血小鼠外周血象、骨髓有核细胞(BMC)数的影响；通过酶联免疫吸附技术(ELISA)检测了骨髓抑制贫血小鼠骨髓及血清Flt3配体(Flt3 ligand, FL)的水平。结果：高剂量维肝力对粒细胞系、红细胞系、血红蛋白(Hb)及BMC均有显著的提高作用,中、低剂量维肝力对血小板促进作用更明显;维肝力组骨髓及血清FL水平均低于贫血组，其中高、中剂量组FL水平下降最为显著。结论：维肝力对放、化疗所引起的小鼠骨髓抑制能起到一定的缓解作用。     

环磷酰胺所致血虚证小鼠骨髓CD34~ 细胞的变化

目的 :研究环磷酰胺 (CTX)诱发的血虚证小鼠骨髓中干祖细胞的数量和骨髓细胞周期的动态变化。方法 :采用CTX大小两个剂量分别对两组小鼠进行腹腔注射制作小鼠血虚证模型。在不同时相点 ,双荧光标记测定CD34 细胞在骨髓有核细胞中的比例 ,骨髓细胞固定后PI染色测定细胞周期。结果 :①注射CTX后CD34 细胞在骨髓有核细胞中的比例下降后迅速上升 ,然后又下降。②注射CTX后骨髓细胞增殖受抑 ,然后动员骨髓细胞增殖 ,但药后 10d细胞增殖又受抑。与CD34 细胞在骨髓有核细胞中比例的变化规律有一致性。结论 :CTX可以动员骨髓造血干祖细胞进入细胞周期而增殖 ,增加了骨髓中造血干祖细胞的数量 ,但此过程加速了干祖细胞池的耗竭 ,从而对骨髓造成更大损伤 ,这可能是CTX诱发小鼠血虚证的机理之一。     

中药促进骨髓造血机理的实验研究进展

从造血生长因子、造血基质细胞、造血微循环及细胞凋亡等方面探讨了中药促进骨髓造血的机理 ,为中药的应用提供科学依据。     

芍药苷对人骨髓基质细胞HFCL蛋白质表达的作用

目的检测芍药苷对人骨髓成纤维样基质细胞系HFCL增殖及蛋白质表达的影响,探讨芍药苷补血作用的分子机制。方法采用流式细胞术和蛋白质组学方法测定芍药苷对HFCL细胞周期及蛋白质表达的影响。结果芍药苷能促进HFCL由G0/G1期进入S期,提高增殖指数,使HFCL的9种蛋白质表达上调,5种下调。上调的蛋白质有Ras相关核蛋白、核纤层蛋白A/C、异柠檬酸脱氢酶、磷酸丙糖异构酶、ATP合酶、核蛋白体蛋白质P2和CCT;下调的蛋白质有人类cc趋化因子和Bax。结论芍药苷能促进HFCL增殖,作用于HFCL多个靶点,促进细胞结构蛋白质的合成和蛋白质分子伴侣的表达,促进HFCL的能量代谢,抑制HFCL凋亡,间接发挥补血作用。     

恩替卡韦治疗骨髓干细胞移植乙肝肝硬化患者HBV血清检测的水平变化

目的探讨骨髓干细胞(BMSCs)移植的乙肝肝硬化患者恩替卡韦治疗72周后血清HBsAg、HBeAg及HBV DNA定量变化的相关性。方法对22例乙肝肝硬化患者给予恩替卡韦抗病毒同时,进行BMSCs移植,在治疗前及治疗中(12,24,48及72周)收集血清,用PCR法进行HBV DNA检测,以COBAXE全自动电化学免疫发光分析系统进行血清HBsAg、HBeAg定量检测。分析恩替卡韦治疗BMSCs移植乙肝肝硬化患者过程中HBsAg、HBeAg及HBV DNA定量值变化的相关性。结果 22例乙肝肝硬化患者治疗过程中HBeAg及HBVDNA定量值均呈下降趋势,呈显著正相关(R=-0.63,P<0.01);随着时间的延长,HBeAg与HBV DNA定量值持续下降,血清HBsAg与HBeAg定量值变化呈负相关关系,(R=-0.487,P<0.01);HBsAg与HBV DNA定量值变化也呈负相关关系(R=-0.358,P<0.01)。结论恩替卡韦治疗BMSCs移植乙肝肝硬化患者中,HBeAg与HBV DNA定量值呈显著相关性,HBsAg定量变化与HBeAg及HBV DNA定量变化呈负相关。