17β-雌二醇对体外培养破骨细胞凋亡及其骨吸收调节作用

目的 观察17β-雌二醇对体外培养破骨细胞凋亡率的影响及其与骨吸收功能的关系。方法 在培养液中加入17β-雌二醇，透射电镜（transmission electron microscopy，TEM）观察破骨细胞内超微结构的改变，流式细胞仪（flow cytometry，FCM）观察不同浓度的17β-雌二醇在不同时间段对破骨细胞凋亡率的影响，同时用扫描电镜（scanning electron microscopy，SEM）观察破骨细胞在骨片上形成骨吸收陷窝数目及面积的变化。结果 17β-雌二醇可增加破骨细胞的凋亡率，这种作用具有剂量、时间依赖效应，同时，破骨细胞在骨片上形成的骨吸收陷窝的数目和面积减少。结论 17β-雌二醇可促进破骨细胞凋亡，从而抑制破骨细胞的骨吸收功能。     

β_2肾上腺素能受体激动剂/拮抗剂对大鼠OB IGF-Ⅰ受体mRNA表达的影响

目的通过研究β2肾上腺素能受体(ADRβ2R)激动剂/拮抗剂对大鼠成骨细胞(OB)成骨性指标胰岛素样生长因子-Ⅰ受体(IGF-ⅠR)mRNA表达的调控作用,探讨交感神经系统(SNS)对骨代谢影响的分子机制。方法取24 h内出生的新生SD大鼠的颅盖骨OB进行体外培养,通过细胞形态学观察、碱性磷酸酶重氮盐法染色和矿化结节茜素红S染色法来鉴定OB。将传至第三代的OB分成A组(ADRβ2R激动剂)、B组(ADRβ2R拮抗剂)和C组(对照组),分别用不同浓度的ADRβ2R激动剂(salbutamol)或ADRβ2R拮抗剂(ICI-118551)对OB处理24 h、72 h和96 h,检测IGF-ⅠR mRNA的表达变化,观察时间效应和剂量效应的关系。结果 ADRβ2R拮抗剂ICI-118551可以上调OB上IGF-ⅠR mRNA的表达;而ADRβ2R激动剂salbutamol则下调IGF-ⅠR mRNA的表达。两种调节作用均具有时间剂量依赖性。说明β2肾上腺素能受体(ADRβ2R)激动剂/拮抗剂可以通过作用于SD大鼠OB上的ADRβ2R而影响OB的骨形成指标IGF-ⅠR mRNA的表达,间接影响了骨形成因子IGF-Ⅰ的促进骨形成能力,从而调节骨代谢。     

淫羊藿苷对破骨细胞活性的影响

目的:观察淫羊藿苷对破骨细胞骨吸收及凋亡的影响,探讨淫羊藿苷的抗骨质疏松作用机制。方法:体外分离、培养兔破骨细胞,与玻片及骨磨片共同培养,用10-7、10-6、5×10-6、10-5mol/L浓度的淫羊藿苷刺激破骨细胞,倒置相差显微镜下观察活体细胞、HE染色、TRAP染色及骨吸收陷窝甲苯胺蓝染色,鉴定破骨细胞,并进行骨吸收陷窝计数和面积测量,吖啶橙染色观察凋亡破骨细胞所占的比例。结果:与空白对照组比较,10-6、5×10-6、10-5mol/L浓度的淫羊藿苷组破骨细胞凋亡率均明显增高,骨吸收陷窝数目、面积明显减少,随浓度增加抑制作用增强,差异有显著性意义(P<0.05)。结论:淫羊藿苷可诱导破骨细胞凋亡,抑制骨吸收,并随浓度增加抑制作用增强。     

两种不同降钙素对体外培养破骨细胞影响的实验研究

目的 观察不同浓度鳗鱼降钙素与鲑鱼降钙素对体外分离培养的破骨细胞数量和活力的影响.方法 从新生幼兔四肢长骨中机械分离出破骨细胞,分别接种于盖玻片及骨薄片上,加入两种不同种类的降钙素,抗酒石酸酸性磷酸酶(TRAP)染色观察盖玻片上破骨细胞的数量和形态,JD801图像分析软件分析骨片上骨吸收陷窝的面积.结果 两种降钙素的各个浓度组(分别是10-12 mol/L、10-10 mol/L、10-8 mol/L)均对贴壁生长的破骨细胞数量有明显抑制作用,并呈剂量相关性.骨吸收陷窝面积分析结果显示,两类降钙素均对破骨细胞吸收功能有明显抑制作用,呈剂量相关性.无论是盖玻片培养还是骨薄片培养,益钙宁和密盖息各相同浓度组间两两对照比较差异均无显著性 (P>0.05).结论 两种不同种类降钙素对体外培养的破骨细胞数量和骨吸收活性的抑制能力相仿.     

重组人骨保护素对体外培养兔破骨细胞的影响

目的观察重组人骨保护素(recombinanthumanosteoprotegerin,rhOPG)对体外培养兔破骨细胞(osteoclast,OC)生存和功能的影响。方法将rhOPG用于干预从新生兔分离出的OC,于干预后1、3、7d取细胞玻片、皮质骨片进行HE、Giemsa、甲苯胺蓝染色,并观察抗酒石酸酸性磷酸酶(TRAP)染色阳性细胞和骨片吸收陷窝,扫描电镜进一步观察吸收陷窝形态。结果分离培养的兔OC多核形态明显;rhOPG对TRAP阳性OC生存的影响,1、3d结果差别不明显,7d的结果差异有显著性(P<0.05);rhOPG能够明显地抑制OC在骨片上形成吸收陷窝,三个时点计数结果差异均有显著性(P<0.01)。结论rhOPG可明显地抑制体外培养兔OC的骨吸收功能。     

阿仑膦酸钠抑制破骨细胞体外吸收的研究

本研究利用体外分离培养的兔破骨细胞,观察第三代双膦酸盐阿仑膦酸钠（ａｌｅｎｄｒｏｎａｔｅ）对骨吸收的抑制作用。将ａｌｅｎｄｒｏｎａｔｅ加入培养液中使其最终浓度为０Ｍ,１μＭ,１０μＭ,１００μＭ。同时观察两组用ａｌｅｎｄｒｏｎａｔｅ盐溶液１００μＭ,５０μＭ浸泡的骨片对破骨细胞吸收功能的影响。用倒置光相差显微镜在不同时间点观察计数骨吸收陷窝并拍照。结果随着培养液内ａｌｅｎｄｒｏｎａｔｅ浓度增高,骨吸收陷窝数减少,面积亦减少。而用ａｌｅｎｄｒｏｎａｔｅ浸泡过的骨片上未见骨吸收陷窝。说明ａｌｅｎｄｒｏｎａｔｅ具有抑制破骨细胞体外骨吸收的作用。     

血小板衍生生长因子－BB对破骨细胞功能影响的实验研究

目的探讨血小板衍生生长因子 -BB(platelet-derivedgrowthfactor-BB,PDGF -BB)对破骨细胞生物学功能的影响。方法 (1)利用酶消化法分离培养成人破骨细胞 ;(2)应用透射电子显微镜方法观察破骨细胞对PDGF -β 受体的表达 ;(3)在经过纯化的破骨细胞上清液中施加重组人基因PDGF -BB ,利用酶动力学方法测定培养上清液中的酸性磷酸酶 (ACP)和抗酒石酸酸性磷酸酶(TRAP)的活性 ;(4)通过Leica图像分析仪观察在PDGF -BB作用下 ,破骨细胞所形成骨吸收陷窝的数目与面积。结果 (1)破骨细胞膜上有胶体金颗粒沉着 ;(2)破骨细胞培养上清液中的ACP和TRAP活性随着PDGF -BB浓度的升高而增加 ,分别从(1.85±0.13)u/L和(1.73±0.15)u/L(对照组 )增加至(2.86±0.15)u/L和(2.75±0.24)u/L(40μg/L组 ,P<0.01) ;(3)在PDGF -BB的作用下 ,破骨细胞所形成的骨吸收陷窝的面积从(435.08±237.50)μm2(对照组 )增高至(630.26±240.64)μm2 (40μg/L组 ,P<0.01) ,每个骨片上骨吸收陷窝的数目亦从14.00±1.41增加为26.00±2.00(P<0.05或P<0.01)。结论PDGF -BB通过与PDGF -β受体结合 ,可直接促进破骨细胞的骨吸收功能     

波尔定对破骨细胞分化及骨吸收功能的影响

目的研究波尔定对NF-κB受体活化因子配体(RANKL)诱导的小鼠单核细胞(RAW264.7)向破骨细胞(OC)分化及骨吸收功能的影响。方法采用RANKL(50 ng/ml)诱导RAW264.7细胞向OC分化,不同浓度波尔定(1、10、20、40、80μmol/L)联合分化培养。培养第4d采用抗酒石酸酸性磷酸酶(TRAP)染色,计算OC样细胞数量;比色法测定TRAP活性;Real-time PCR法检测各组OC标志性基因核因子κB受体活化因子(RANK)、活化T细胞核因子1(NFATc1)和c-fos基因的表达;CCK-8法检测波尔定对OC的毒性。培养至第9d,采用甲苯胺蓝染色并用Leica Qwin图像分析系统计算骨吸收陷窝面积。结果与对照组比较,TRAP阳性OC数量、TRAP活性、OC标志性基因RANK、NFATc1和c-fos的表达量及骨陷窝面积,随着波尔定浓度增加而减少,与对照组比较,20μmol/L、40μmol/L、80μmol/L波尔定可明显减少OC数量和TRAP活性(P0.05,P0.01),下调OC标志性基因RANK、NFATc1和c-fos的表达量及骨陷窝面积(P0.05),但以上浓度波尔定对OC均未产生毒性。结论波尔定对RANKL诱导的RAW264.7细胞向OC的分化及骨吸收功能具有抑制作用。     

阿司匹林对大鼠破骨细胞分化成熟和骨吸收活性的影响

目的 研究不同浓度阿司匹林对体外培养大鼠破骨细胞(Osteoclast,OC)分化成熟及骨吸收活性的影响.方法 建立由核激活因子受体配体(receptor activator of NF-κB ligand,RANKL)和巨噬细胞集落刺激因子(Macrophage colony stimulating factor,M-CSF)共同作用的大鼠破骨细胞骨髓诱导体系,将雌激素(10-6 mmol/L)和不同浓度的阿司匹林(0.25 mmol/L、0.5 mmol/L、1.0 mmol/L、1.5 mmol/L)分别作用于破骨细胞.诱导培养后分别对破骨细胞进行抗酒石酸酸性磷酸酶(The tartrate-resistant acid phosphatase,TRAP)染色,观察细胞形态,并计数破骨样细胞数量;将各组破骨细胞接种于骨磨片上,建立破骨细胞-骨磨片活性分析模型,于不同时间点对骨磨片进行光镜和扫描电镜观察,分析计算骨吸收陷窝面积.结果 与正常对照组相比,雌激素组破骨细胞数量和骨吸收陷窝面积低于正常对照组,差异有统计学意义(P＜0.05);且随着阿司匹林浓度的增加,阿司匹林组TRAP阳性多核破骨细胞数量、骨吸收陷窝面积逐渐减少直至消失,差异有统计学意义(P＜0.05).与雌激素组相比,低浓度阿司匹林组(0.25mmol/L)没有明显差异;但中、高浓度阿司匹林实验组(0.5mmol/L、1.0mmol/L、1.5mmol/L)破骨细胞数量和骨吸收陷窝面积减少,差异有统计学意义(P＜0.05).结论 阿司匹林对破骨细胞的分化成熟及骨吸收功能有抑制作用,且呈剂量依赖性,从而具有抗骨质疏松的作用.     

转移生长因子β1对体外培养破骨细胞功能影响的实验研究

目的探讨转移生长因子β1(TGF-β1)对体外培养破骨细胞功能影响的机制.方法酶动力学法测定培养基中酸性磷酸酶和抗酒石酸酸性磷酸酶活性;共聚焦激光扫描显微镜观察体外培养破骨细胞内氢离子随TGF-β1浓度的变化情况;Leica Quantimet 500图像分析系统对骨吸收陷窝进行图像分析;扫描电镜观察骨吸收陷窝变化.结果随着TGF-β1浓度的升高,处理组与对照组ACP和TRAP的活性分别从(1.71±0.33U)/L和(1.41±0.29)U/L降低到(1.32±0.21)U/L和(1.01±0.11)U/L(均为P＜0.01),骨吸收陷窝的数目与面积从(16.67±1.35)个/片和(582.24±178.68)μm2减少到(13.29±1.47)个/片与(442.38±148.27)μm2,处理组与对照组比较差异具有显著性(P＜0.01).破骨细胞相对荧光值无显著性差异,表明TGF-β1对体外培养破骨细胞分泌H+无明显影响.结论TGF-β1虽然不能抑制氢离子释放,但能通过改变酸性磷酸酶和抗酒石酸酸性磷酸酶活性,进而直接抑制体外培养破骨细胞的骨吸收功能.     

应激激素肾上腺素激活ERK1/2促进人肝癌细胞生长

目的 探讨应激激素肾上腺素(EPI)对人肝癌细胞生长的影响及其作用机制.方法 采用噻唑蓝(MTT)、逆转录-聚合酶链反应(RT-PCR)和Western blot等方法,分析EPI对人肝癌HepG2和MHCC97H细胞生长的影响,与α1-/β2-肾上腺素受体(α1-/β2-ARs)、腺苷环化酶/蛋白激酶A(AC/PKA)、促分裂原活化蛋白激酶/细胞外信号调节激酶1/2(MAPK/ERK1/2)和磷脂酸激醇-3-激酶/蛋白激酶B(PI3K/AKT/PKB)的关系.结果癌细胞增殖与EPI的作用呈剂量和时间依赖关系(P<0.05),10μmol/L EPI作用24～48 h时其增殖能力最强[HepG2/(215±4)%和MHCC97H/(207±10)%],而正常肝细胞L-02的变化不明显(P>0.05).在HepG2和MHCC97H细胞中,α1-AR蛋白表达仅为L-02的30%和34%,而β2-AR蛋白表达上升至331%和309%.β2-AR阻滞剂ICI 118551可抑制EPI的促增殖作用.β-AR激动剂异丙肾上腺素(ISO,10μmol/L)具有类似EPI的促进增殖作用,其作用可分别被ICI118551和MEK抑制剂U0126显著抑制,被PKA抑制剂H-89和PI3K抑制剂LY294002部分抑制.ISO孵育15 min～8 h,在HepG2中p-ERK1/2蛋白水平上升3.49/3.02倍,在MHCC97H中上升3.15/2.73倍,该作用可被ICI 118551和U0126所阻断.结论 人肝癌HepG2和MHCC97H细胞过表达β2-AR,EPI通过β2-AR激活ERK1/2依赖性和非依赖性信号通路促进癌细胞的生长,其中MAPK/ERK1/2信号通路可能起主要作用.     

Polarized osteoclasts put marks of tartrate-resistant acid phosphatase on dentin slices--a simple method for identifying polarized osteoclasts

Osteoclasts form ruffled borders and sealing zones toward bone surfaces to resorb bone. Sealing zones are defined as ringed structures of F-actin dots (actin rings). Polarized osteoclasts secrete protons to bone surfaces via vacuolar proton ATPase through ruffled borders. Catabolic enzymes such as tartrate-resistant acid phosphatase (TRAP) and cathepsin K are also secreted to bone surfaces. Here we show a simple method of identifying functional vestiges of polarized osteoclasts. Osteoclasts obtained from cocultures of mouse osteoblasts and bone marrow cells were cultured for 48 h on dentin slices. Cultures were then fixed and stained for TRAP to identify osteoclasts on the slices. Cells were removed from the slices with cotton swabs, and the slices subjected to TRAP and Mayer's hematoxylin staining. Small TRAP-positive spots (TRAP-marks) were detected in the resorption pits stained with Mayer's hematoxylin. Pitted areas were not always located in the places of osteoclasts, but osteoclasts existed on all TRAP-marks. A time course experiment showed that the number of TRAP-marks was maintained, while the number of resorption pits increased with the culture period. The position of actin rings formed in osteoclasts corresponded to that of TRAP-marks on dentin slices. Immunostaining of dentin slices showed that both cathepsin K and vacuolar proton ATPase were colocalized with the TRAP-marks. Treatment of osteoclast cultures with alendronate, a bisphosphonate, suppressed the formation of TRAP-marks and resorption pits without affecting the cell viability. Calcitonin induced the disappearance of both actin rings and TRAP-marks in osteoclast cultures. These results suggest that TRAP-marks are vestiges of proteins secreted by polarized osteoclasts.     

两种方法培养大鼠破骨细胞的比较研究

目的比较分析直接分离培养的Wistar大鼠破骨细胞（osteoclast，OC）和诱导培养的Wistar大鼠破骨样细胞（osteoclast-like cell，OLC）的形态和功能的差异，为体外药物干预试验奠定基础。方法采用两种培养法，即从新生（24h内）的Wistar大鼠的四肢长骨骨髓腔内壁机械分离成熟OC直接培养和10^-8mol／L的1，25（OH）2D3诱导4周龄Wistar大鼠骨髓单核细胞形成OLC的方法，对获得的OC／OLC进行形态和破骨功能观察。结果两种方法都培养出了抗酒石酸酸性磷酸酶（tartrate resistant acid phosphatase，TRAP）染色阳性的多核细胞，诱导法获得的破骨细胞数量较多（P〈0．05）。成熟OC与诱导获得的OLC形态特征相似，但后者在骨片上形成的陷窝较小而浅。结论直接分离培养法可获得骨吸收功能较活跃的OC，但数目较少，适合骨吸收功能分析、破骨迁移黏附、凋亡研究及单细胞分子生物学研究。1，25（OH）2D3诱导鼠骨髓单核细胞形成的OLC数量较多，但骨吸收功能较差，适合用于破骨细胞分化发育过程的研究。     

Deletion of β-adrenergic receptor 1, 2, or both leads to different bone phenotypes and response to mechanical stimulation

As they age, mice deficient for the β2-adrenergic receptor (Adrb2(-/-) ) maintain greater trabecular bone microarchitecture, as a result of lower bone resorption and increased bone formation. The role of β1-adrenergic receptor signaling and its interaction with β2-adrenergic receptor on bone mass regulation, however, remains poorly understood. We first investigated the skeletal response to mechanical stimulation in mice deficient for β1-adrenergic receptors and/or β2-adrenergic receptors. Upon axial compression loading of the tibia, bone density, cancellous and cortical microarchitecture, as well as histomorphometric bone forming indices, were increased in both Adrb2(-/-) and wild-type (WT) mice, but not in Adrb1(-/-) nor in Adrb1b2(-/-) mice. Moreover, in the unstimulated femur and vertebra, bone mass and microarchitecture were increased in Adrb2(-/-) mice, whereas in Adrb1(-/-) and Adrb1b2(-/-) double knockout mice, femur bone mineral density (BMD), cancellous bone volume/total volume (BV/TV), cortical size, and cortical thickness were lower compared to WT. Bone histomorphometry and biochemical markers showed markedly decreased bone formation in Adrb1b2(-/-) mice during growth, which paralleled a significant decline in circulating insulin-like growth factor 1 (IGF-1) and IGF-binding protein 3 (IGF-BP3). Finally, administration of the β-adrenergic agonist isoproterenol increased bone resorption and receptor activator of NF-κB ligand (RANKL) and decreased bone mass and microarchitecture in WT but not in Adrb1b2(-/-) mice. Altogether, these results demonstrate that β1- and β2-adrenergic signaling exert opposite effects on bone, with β1 exerting a predominant anabolic stimulus in response to mechanical stimulation and during growth, whereas β2-adrenergic receptor signaling mainly regulates bone resorption during aging.     

The effects of the cathepsin K inhibitor odanacatib on osteoclastic bone resorption and vesicular trafficking

Odanacatib (ODN) is a selective, potent and reversible inhibitor of cathepsin K (CatK) that inhibits bone loss in postmenopausal osteoporosis. Evidence from osteoclast (OC) formation from bone marrow of CatK^−/− mice or human OC progenitors treated with ODN, demonstrated that CatK inhibition has no effect on osteoclastogenesis or survival of OCs. Although having no impact on OC activation, ODN reduces resorption activity as measured by CTx release (IC₅₀ = 9.4 nM) or resorption area (IC₅₀ = 6.5 nM). While untreated cells generate deep trail-like resorption lacunae, treated OCs form small discrete shallow pits. ODN leads to significant accumulation of intracellular vesicles intensely stained for CatK and TRAP. CatK (+) vesicles localize toward the basolateral and functional secretory membranes of the polarized OC and TRAP(+) vesicles evenly distribute in the cytoplasm, suggesting that ODN disrupts multiple vesicular trafficking pathways. Intracellular levels of both precursor and mature TRAP were increased by 2-fold and the pre-pro and mature CatK by 6- and 2-fold in ODN-treated OCs compared to untreated controls. ODN treated OC accumulates labeled degraded bone matrix proteins in CatK containing vesicles. In summary, ODN treatment inhibits bone resorption by blocking degradation of demineralized collagen in the resorption lacunae, and retarding transcytosis for further processing of degraded proteins.     

皮肤创面愈合过程中上皮间质化机制的研究

目的:探讨β2肾上腺素受体(β2-AR)在皮肤创伤修复上皮间质化(EMT)过程中的作用机制及意义.方法:采用免疫印迹法(Western blot)、逆转录-聚合酶链式反应(RT-PCR)、免疫荧光技术等实验方法,体外观察β2-AR激动剂(ISO)和抑制剂(ICI)对上皮细胞间质化标志物(α-平滑肌肌动蛋白、波形蛋白、snail)表达的影响及创面皮肤上皮化标志物(E-钙黏附素)表达的变化.结果:β 2-AR可通过介导皮肤创面愈合的EMT过程,促进表皮细胞向间质表型转化,且运动和迁移能力在该过程中不断增强.结论:β 2-AR是皮肤上皮间质化的关键分子,其介导的EMT过程是皮肤创面愈合的生物学基础.     

依降钙素对破骨细胞的作用观察

目的 观察了国产降钙素-依降钙素对破骨细胞体外骨吸收功能的影响。并与进口降钙素-益钙宁的作用比较，方法 由10日龄幼兔四肢长骨机械分离破骨细胞于199培养液（含10%NCS 5?S），分别接种于象牙薄切片和培养板，置37℃，5%CO2培养箱中培养，细胞骨架观察采用荧光标记法（罗达明标记的鬼笔环肽），骨片吸收陷窝计数采用甲苯胺蓝染色法，益钙宁为阳性对照，等量培养液为阴性对照，结果 依降钙素组破骨细胞F-actin聚集，变短，骨片培养3d，吸收陷窝计数的结果显示，10^-10mol/L以上浓度依降钙素对破骨细胞吸收功能有明显抑制作用，并呈剂量相关性，10^-1mol/L时的抑制作用高达90%；依降钙素和益钙宁的抑制作用呈高度相关（r=0.99）。结论 依降钙素具有明显的抑制体外培养破骨细胞骨吸收活性的作用，与益钙宁的作用相仿。     

Adhesion patterns and cytoskeleton of rabbit osteoclasts on bone slices and glass

The ability of osteoclasts (OC) to migrate and resorb bone is thought to be dependent on cytoskeletal function and adhesion. Therefore, we investigated the cytoskeleton and the adhesion patterns of rabbit OC on glass and on devitalized bone slices, using specific antibodies to cytoskeletal elements and fluorescence and interference reflection microscopy. Microtubules (MT) were similar in OC on both substrata, and appeared in a pattern typical of that described for many cells. Multiple centriolar complexes were observed in most OC, either as one large aggregate in the center of the cell or dispersed singly or in small aggregates close to individual nuclei. Staining of microfilaments (MF) was similar on both substrata and appeared primarily as an F-actin network. MF distribution was different in OC associated with resorption lacunae with intense staining over those regions. In the OC on glass, high F-actin staining was detectable at the periphery in dots and rosette-like structures, which also stained for vinculin. The adhesion patterns indicated that OC on glass do not make large focal contacts, but appear to make a few tiny focal contacts that are not associated with the rosette-like structures. Most of the undersurface of the OC appeared either to be involved in close contacts or to be separated by distances of greater than 100 nm from the substratum. These studies indicate that the MF distribution and the adhesion patterns of rabbit OC are typical of motile cells, that the distribution of the cytoskeleton of rabbit OC on glass and on bone slices is similar, and that MF may be involved in the morphological changes associated with resorption.     

BM210955对破骨细胞骨吸收的抑制作用

细胞水平研究国产双磷酸盐药物－ＢＭ２１０９５５的骨吸收抑制作用。由１日龄ＳＤ大鼠四肢长骨分离破骨细胞（ＯＣ）并接种于牛皮质骨薄切片上,在不同浓度ＢＭ２１０９５５作用下培养,定时取骨片作ＴＲＡＰ免疫组化染色,计数阳性多核细胞后,经超声去除细胞后作甲苯胺蓝染色,光镜下作吸收陷窝计数,数据以ｘ±ｓ表示,并与对照比较作ｔ检验。结果显示：①ＢＭ２１０９５５能降低体外培养ＯＣ的数目,１０－８ｍｏｌ／Ｌ组的ＴＲＡＰ阳性多核细胞较对照组减少７３％,差异具有非常显著性,Ｐ＜０．０１。②ＢＭ２１０９５５抑制ＯＣ的陷窝形成能力,１０－１０、１０－８ｍｏｌ／Ｌ组的抑制率分别为７６．３２％和８７．９９％,均与对照有明显差异,Ｐ＜０．０１。③上述结果与剂量有关,剂量越高,抑制效应越明显