锡的示波极谱法测定

在Ｈ２ＳＯ４－Ｈ２Ｃ２Ｏ４介质中,用单扫描示波极谱法,Ｓｎ（ＩＶ）－ＤＰＧ体系可产生灵敏的络合吸附波。锡浓度在８～４００μｇ／ｌ范围内与峰高呈线性关系,检测限为２μｇ／ｌ,并用于硬聚氯乙烯饮用水管材和管件锡及水中微量锡的测定,结果满意,回收率在８９～１０３％,相对标准偏差ＲＳＤ％为０．５２％～５．６０％之间。     

补肾中药对去卵巢大鼠骨组织ER、OPG表达的促进作用

通过检测去卵巢大鼠给予补肾中药后腰椎内ERmRNA及OPG mRNA的表达,进一步探讨补肾中药防治骨质疏松的分子机制.将雌性3月龄SD大鼠48只,随机分为卵巢切除 补肾中药防治(HDP),卵巢切除 雌激素(倍美力)防治(ER),骨质疏松(卵巢切除)(OVX)及正常对照组(SHAM)4组,每组12只.术后12周处死大鼠,取L3椎体,异硫氰酸胍一步法提取总RNA,嵌套式反转录聚合酶链反应(NRT-PCR),扩增待检测各组ERmRNA及OPGmRNA的表达.同时取L4椎体,行骨组织形态计量学检测.结果显示HDP组ER mRNA及OPG mRNA阳性表达率分别为75.0%和66.7%;EP组ER mRNA及OPG mRNA阳性表达率均为83.3%;OVX组各例ER mRNA均无表达,2例OPG mRNA呈弱阳性表达(阳性率为16.7%);SHAM组呈ER mRNA及OPG mRNA阳性表达.χ2检验,HDP组与ER组比较ER mRNA及OPG mRNA阳性表达率均无显著性差异(P＞0.05);该两组与OVX组比较,ER mRNA及OPG mRNA阳性表达率均有高度显著性差异(P＜0.01);而与SHAM组比较,HDP组ERmRNA,ER组ER mRNA及OPG mRNA阳性表达率无显著性差异(P＞0.05),但HDP组OPG mRNA阳性表达率有显著性差异(0.01＜P＜0.05).骨组织形态计量学检测,OVX组骨体积、骨小梁厚度及骨小梁数目均比SHAM组显著减少,而骨小梁间隔显著增大.同EP组相似,HDP组骨体积、骨小梁厚度显著增加,虽也使骨小梁间隔稍有减少,但与OVX组比较无统计学差异.表明补肾中药通过直接作用于骨组织中ER,增强ER mRNA的表达,并刺激OPG的分泌,从而有望替代雌激素,对去卵巢大鼠的骨质疏松有明显的防治作用.     

TGF-β对成骨细胞增殖、分化及OPG基因mRNA表达的影响

目的 观察不同剂量TGF—β对成骨细胞增殖、分化及OPGmRNA基因表达的影响，明确其作用机理。方法 取新生24h内的Wistar大鼠颅盖骨分离成骨细胞，在TGF—β为0μg／L、1μg／L、10μg／L、100μg／L的浓度中培养。对细胞的生长及钙结节的形成观察其生长状态，经MTT法测定细胞增殖，细胞分化由碱性磷酸酶(ALP)和骨钙素(OCN)水平测定得到。用RT—PCR技术测定TGF—β对成骨细胞OPG基因mRNA表达的影响。结果 成骨细胞在7d可铺满瓶壁，30d可形成钙结节，TGF—β可促进成骨细胞的增殖、ALP和OCN的分泌，以TGF—β为1μg／IJ时作用明显。TGF—β可促进OPG基因的表达在为1μg／L时作用明显。结论 1μg／L可促进大鼠成骨细胞的增殖、分化，促进成骨细胞OPG基因mRNA的表达。TGF—β可能通过影响OPG而调节成骨细胞、破骨细胞的平衡，使骨重建。     

Elevated soluble receptor activator of nuclear factor-κB ligand and reduced bone mineral density in patients with adolescent idiopathic scoliosis

Generalized low bone mass and osteopenia in both axial and peripheral skeleton in adolescent idiopathic scoliosis (AIS) have been reported in literature. However, the exact mechanisms and causes of the bone loss in AIS are not identified yet. Therefore, this study examined the relationship between serum concentration of soluble receptor activator of nuclear factor-κB ligand (RANKL), serum level of osteoprotegerin (OPG) and bone mass in 72 patients with AIS and compared to those of 64 age- and gender-matched healthy controls. The mean lumbar spinal bone mineral density (LSBMD) and femoral neck BMD (FNBMD) in patients with AIS were decreased compared with that in control individuals, respectively (P = 0.0029 and P = 0.0192, respectively). The mean RANKL and RANKL to OPG ratio in patients with AIS were increased compared with that in control subjects, respectively (P = 0.0004 and P = 0.0032, respectively). The RANKL and RANKL to OPG ratios were negatively correlated to the LSBMD and serum OPG levels in both groups. Serum OPG levels were positively correlated to the LSBMD and FNBMD in both groups. These findings mean that the imbalance and the disturbed interaction of RANKL and OPG may be an important cause and pathogenesis in reduced BMD in AIS.     

刺老苞根皮黄酮类化合物对MC31E-E1成骨细胞OPG和RANKL mRNA基因表达影响的实验研究

目的:探讨刺老苞根皮黄酮类化合物对MC3T3-E1成骨细胞骨保护素(OPG)和核因子κB受体激活因子配体(RANKL)mRNA基因表达的影响.方法:体外培养MC313-E1成骨细胞,分别加入含0mol·L-1、10-8mol·L-1、10-7mol·L-1、10-6mol·L-1刺老苞根皮黄酮类化合物的培养液,48h及96h后分别提取细胞总mRNA,用RT-PCR方法分析OPG/RANKL的mRNA的表达,进行半定量分析.结果:培养48h后,与对照组比较,刺老苞根皮黄酮类化合物增强了OPGmRNA的表达,有显著性差异(P<0.05或P<0.01),VOPG/VRANKL比值增大;96h后,刺老苞根皮黄酮类化合物明显增强了OPG mRNA的表达,各浓度组与对照组相比,均有显著性差异(P<0.05或P<0.01),各浓度组VOPG/VRANKL比值增大,有显著性差异(P<0.05或P<0.01).结论:刺老苞根皮黄酮类化合物可能通过增加成骨细胞OPG的表达来抑制破骨细胞的分化和成熟,从而抑制骨吸收,达到治疗骨质疏松症的目的.     

Differential effects of biologic versus bisphosphonate inhibition of wear debris‐induced osteolysis assessed by longitudinal micro‐CT

Aseptic loosening of total joint replacements is caused by wear debris‐induced osteoclastic bone resorption, for which bisphosphonates (BPs) and RANK antagonists have been developed. Although BPs are effective in preventing metabolic bone loss, they are less effective for inflammatory bone loss. Because this difference has been attributed to the antiapoptotic inflammatory signals that protect osteoclasts from BP‐induced apoptosis, but not RANK antagonists, we tested the hypothesis that osteoprotegerin (OPG) is more effective in preventing wear debris‐induced osteolysis than zoledronic acid (ZA) or alendronate (Aln) in the murine calvaria model using in vivo micro‐CT and traditional histology. Although micro‐CT proved to be incompatible with titanium (Ti) particles, we were able to demonstrate a 3.2‐fold increase in osteolytic volume over 10 days induced by polyethylene (PE) particles versus sham controls (0.49 ± 0.23mm³ versus 0.15 ± 0.067mm³; p < 0.01). Although OPG and high‐dose ZA completely inhibited this PE‐induced osteolysis (p < 0.001), pharmacological doses of ZA and Aln were less effective but still reached statistical significance (p < 0.05). Traditional histomorphometry of the sagital suture area of calvaria from both Ti and PE‐treated mice confirmed the remarkable suppression of resorption by OPG (p < 0.001) versus the lack of effect by physiological BPs. The differences in drug effects on osteolysis were largely explained by the significant difference in osteoclast numbers observed between OPG versus BPs in both Ti‐ and PE‐treated calvaria; and linear regression analyses that demonstrated a highly significant correlation between osteolysis volume and sagittal suture area versus osteoclast numbers (p < 0.001). © 2008 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 26:1340–1346, 2008     

骨保护素及其配体在牙正畸中作用的研究进展

骨保护素(OPG)是生理性的抑制破骨细胞性骨吸收的因子,而骨保护素配体(OPGL)是能直接诱导破骨细胞分化发育并参与破骨细胞功能调节的细胞因子。牙周膜细胞可表达OPG和OPGL,揭示了在正畸牙移动中,牙周膜细胞可能是通过OPG/OPGL系统来调节牙槽骨代谢的。OPG和OPGL在多种骨病和牙正畸的治疗中具有很大潜力,具有良好的临床应用前景。     

In vivo expression of RANKL in the rat dental follicle as determined by laser capture microdissection

Tooth eruption is a localized event in which many of the genes required for eruption are expressed in the dental follicle. A major function of the follicle is to recruit mononuclear cells for osteoclastogenesis such that the alveolar bone can be resorbed. Osteoclastogenesis is primarily regulated by receptor activator of nuclear factor-kappa B ligand (RANKL), colony-stimulating factor-one (CSF-1) and osteoprotegerin (OPG). In the rat first mandibular molar, osteoclastogenesis is maximal at day 3 and CSF-1 is maximally expressed in the follicle at this time whereas OPG expression is reduced. Whether or not RANKL is expressed in vivo in the follicle is controversial, however. It is critical to determine this because others have shown that in partially-rescued mice null for RANKL, teeth do not erupt. This suggests that RANKL should be expressed in the follicle for eruption to occur. Thus, to precisely determine if RANKL is expressed in the follicle in vivo, laser capture microdissection (LCM) was used to excise dental follicle tissue from frozen sections followed by RNA isolation and RT-PCR. The results show that RANKL is expressed in the dental follicle at days 1-9 postnatally. The technique was confirmed by controls showing that LCM isolates of the follicle, and alveolar bone, express OPG. Also, LCM isolates of alveolar bone were positive for RANKL. Thus, RANKL has now been shown to be expressed in the follicle and it is probable that interactions between it, CSF-1 and OPG regulate locally the osteoclastogenesis needed for tooth eruption.     

Inhibition of osteoclastogenesis by a phosphodiesterase 4 inhibitor XT-611 through synergistic action with endogenous prostaglandin E2

We examined the effect of a phosphodiesterase 4 (PDE4) inhibitor, 3,4-dipropyl-4,5,7,8-tetrahydro-3H-imidazo[1,2-i]-purin-5-one (XT-611) on osteoclast formation in three different mouse bone-marrow cell (BMC) culture systems. We confirmed that selective inhibitors of PDE4, including XT-611, among several PDE inhibitors decreased osteoclast formation in the BMC culture system. XT-611 also inhibited osteoclast formation in co-culture of mouse bone-marrow stromal cell line ST2 and adherent cell-depleted (ACD)-BMCs. However, it did not inhibit osteoclastogenesis in culture of ACD-BMCs alone in the presence of macrophage-colony stimulating factor (M-CSF) and soluble receptor activator of NF-kappaB ligand (sRANKL). XT-611 significantly increased prostaglandin E(2) (PGE(2)) production from ST2 cells and, in combination with PGE(2), synergistically increased cAMP concentration in osteoclast progenitors. In the ST2 co-culture system, XT-611 did not influence the expression of RANKL, osteoprotegerin and RANK mRNAs. By combined treatment with XT-611 and PGE(2) of ACD-BMCs, osteoclast multinucleation was clearly inhibited with decrease in the expression of calcitonin receptor mRNA, while the expression of RANK and c-fms (an M-CSF receptor) mRNAs was unchanged. These results indicate that the PDE4 inhibitor inhibits osteoclastogenesis by acting on osteoclast progenitors synergistically with PGE(2) secreted from stromal cells, but not by influencing the cell-to-cell interaction between stromal cells and osteoclast progenitors.     

补肾复方含药血清对大鼠成骨细胞OPG、ODF表达的影响

为研究补肾复方对大鼠成骨细胞OPG、ODF表达影响。以补肾活血方、补肾健脾方、龟鹿二仙膏、金匮肾气丸、六味地黄丸、左归丸6种补肾中药复方及生理盐水给体重为250g左右的大鼠灌胃,提取含药血清;采用酶消化法从初生2天SD大鼠头盖骨中分离出成骨细胞,用含药血清及生理盐水进行成骨细胞传代培养;取第2代细胞,培养至细胞铺满培养瓶约80%时,更换含药血清培养液24小时;用含药血清培养液24小时后终止培养,提取总RNA,应用RT-PCR方法观察六种补肾复方对大鼠成骨细胞OPG、ODF表达影响。结果显示补肾活血方、补肾健脾方、金匮肾气丸、六味地黄丸和左归丸含药血清培养的大鼠成骨细胞OPG的表达量高于生理盐水对照组,龟鹿二仙膏含药血清培养的大鼠成骨细胞OPG的表达量低于生理盐水对照组,其中补肾活血方组、补肾健脾方组大鼠成骨细胞OPG表达量明显高于生理盐水对照组,统计学有显著性差异(P<0.05)。6种补肾复方含药血清培养的大鼠成骨细胞ODF表达量均低于生理盐水对照组,其中补肾活血方组、补肾健脾方组和金匮肾气丸组大鼠成骨细胞ODF表达量明显低于生理盐水对照组,统计学有显著性差异(P<0.05)。表明补肾活血方、补肾健脾方含药血清能增加大鼠成骨细胞OPG表达量,降低大鼠成骨细胞ODF表达量。