强筋健骨方对PMOP大鼠骨密度及生物力学影响的实验研究

目的探究强筋健骨方对去势致骨质疏松症大鼠双侧股骨髁骨密度、生物力学特性及骨组织形态学的影响。方法采用经典的去势雌鼠建立绝经后骨质疏松症模型,采用随机数字表随机分为6组,分别为假手术组(Sham)、模型组(OVX)、西药对照组(E2)[12μg/(kg·d)]、"强筋健骨方"低、中、高剂量组(JGG、JGZ、JGD)[5.56 g/(kg·d)、11 g/(kg·d)、22 g/(kg·d)],灌胃给药12周后,观察大鼠股骨生物力学特性、骨矿物含量、双能X线测定双侧股骨髁骨密度,光镜下进行骨组织形态学观察。结果与Sham组相比较,OVX组取材时大鼠股骨矿物含量、股骨髁骨密度、股骨生物力学特性降低,骨组织形态学遭到破坏。与OVX组相比较,E2组及"强筋健骨方"三剂量组均能不同程度改善去势雌鼠的股骨生物力学特性、股骨髁骨密度、骨矿物含量及骨组织微结构。结论强筋健骨方对绝经后骨质疏松大鼠骨密度及生物力学特性破坏有一定改善作用,且在此剂量范围内存在剂量相关性。     

丹酚酸B通过抗氧化作用改善高脂饮食小鼠牙槽骨骨质疏松的实验研究

目的探索丹酚酸B改善高脂饮食引起的小鼠牙槽骨骨量丢失的作用以及可能的作用机制。方法 C57BL/6J雄性小鼠30只,分为3组:正常组、高脂饮食组和丹酚酸B组,每组10只。除正常组外,其他两组给予高脂饮食,丹酚酸B组用丹酚酸B[125 mg/(kg·d)]干预,治疗12 w后,取小鼠牙槽骨,然后分别用双能X射线测量骨密度,HE染色观察骨微结构,免疫组织化学染色分析牙槽骨核因子-κB-p65(nuclear factor-kappa B,NF-κB-p65)、组织蛋白酶K(Cathepsin K)、超氧化物歧化酶(superoxide dismutase,SOD)和NADPH oxidase 4(Nox4)的表达情况。结果丹酚酸B能明显改善高脂饮食诱发的小鼠牙槽骨骨密度下降以及骨微结构破坏。同时,丹酚酸B能上调高脂饮食小鼠牙槽骨SOD的表达,下调Nox4、NF-κB-p65和Cathepsin K的表达。结论丹酚酸B可能通过调节Nox4/SOD/NF-κB/Cathepsin K通路改善高脂饮食引起的氧化应激,从而抑制高脂饮食诱发的小鼠牙槽骨骨量丢失。本研究将为丹参及其有效成分治疗骨质疏松提供科学的实验依据。     

小鼠非酒精性脂肪肝纤维化对骨微结构的影响及姜黄素治疗作用

目的探讨非酒精性脂肪肝(NAFLD)小鼠模型肝纤维化对骨微结构影响及姜黄素的治疗作用。方法 6周龄雌性野生型C57BL/6小鼠30只,随机分为野生小鼠对照组(WT组,n=10),高脂饮食组(HFD组,n=10)和姜黄素治疗组(HFD+Cur组n=10)。对照组给予普通饲料饲养,高脂饮食组给予高脂饮食饲养,姜黄素治疗组除了给予高脂饮食外,在24周时给予姜黄素治疗。各组小鼠于32周末处死,检测小鼠血清生化指标,以及小鼠血清中TNF-α,IL-6,IGF-1和IGFBP-1的水平,肝组织HE染色后进行光镜病理学检测,micro-CT分析骨微结构及骨密度。结果与WT组相比,HFD组血清中AST、IL-6、TNF-α明显升高(P0.05),而体重、TC、ALT、IGF-1、IGFBP-1变化更加明显(P0.01);小鼠肝组织纤维化增生明显,同时胫骨近端参数BV/TV、Tb.Th、Tb.N、Tb.Sp、Conn.D/mm~3、C.Th、v BMD、t BMD及骨干形态与WT组相比差异具有统计学意义(P0.05、P0.01);通过姜黄素治疗后除IGF-1水平明显升高(P0.05),小鼠体重以及血清中其余各指标水平明显下降(P0.05),更接近于WT组(P0.05);肝脏病理学未见明显坏死灶及纤维化,小鼠骨微结构参数及骨干形态较HFD组明显好转(P0.05、P0.01),部分参数接近WT组(P0.05)。结论小鼠非酒精性脂肪肝纤维化期可出现明显骨量丢失,血清中TNF-α、IL-6、IGF-1和IGFBP-1变化在其发病机制中起到重要的作用,其中IGF-1和IGFBP-1作用尤为重要,而姜黄素通过调节这些细胞因子可减轻脂质聚集,增加骨小梁、骨皮质及骨密度。     

格列美脲对高脂饮食ApoE-/-小鼠骨微结构的影响

目的观察格列美脲对高脂饮食喂养的Apo E基因敲除(ApoE~(-/-))小鼠骨微结构的影响。方法以8只野生型C57BL/6小鼠为野生正常组,24只ApoE~(-/-)小鼠随机分为3组,分别为ApoE~(-/-)正常组、ApoE~(-/-)高脂组、格列美脲组。野生正常组和ApoE~(-/-)正常组给予普通饲料饲养,ApoE~(-/-)高脂饮食组和格列美脲组给予高脂饲料,其中格列美脲组小鼠灌服25mg/kg格列美脲,其他各组小鼠均灌服等量去离子水。6w后,处死小鼠,取双侧股骨,分别进行HE染色和Safrain O/Fast Green染色评价骨微结构,利用骨生物力学评价骨强度,利用Cathepsin K表达评价骨吸收程度。结果格列美脲可明显改善高脂饮食喂养的ApoE~(-/-)小鼠骨微结构,提升股骨糖胺聚糖含量,提高股骨第一循环硬度、峰值压力和硬度形变量,降低Cathepsin K表达。结论格列美脲可改善高脂饮食饲养的ApoE~(-/-)小鼠的骨微结构,其作用机理可能与抑制Cathepsin K的表达相关。     

续苓健骨方对去卵巢骨质疏松模型大鼠血液钙磷代谢的影响

目的研究续苓健骨方对去卵巢骨质疏松模型大鼠骨密度、骨微结构和血液钙磷代谢的影响。方法将40只6月龄雌性SD大鼠随机分为假手术组、模型组、续苓健骨方组[12.71 g/(kg·d)]和碳酸钙组[104.19 mg/(kg·d)],假手术组仅切除卵巢周围脂肪,其余组行双侧卵巢切除术后建立骨质疏松模型。药物干预12周后取材,通过双能X线骨密度仪检测左侧胫骨骨密度,经Masson染色观察右侧胫骨组织显微结构,生化检测血清抗酒石酸酸性磷酸酶(TRAP)、血钙和血磷水平。结果与假手术组相比,模型组骨密度显著降低,骨小梁断裂稀疏、排列不规则,TRAP表达增加,血钙含量降低,血磷含量升高。与模型组相比,续苓健骨方组骨密度增加,骨小梁数量增加、排列较规则,TRAP表达降低,血钙含量升高,血磷含量降低。结论续苓健骨方可提高去卵巢骨质疏松大鼠胫骨骨密度并改善骨组织微结构,其机制可能与调控血钙磷代谢稳态有关。     

抗氧化剂对去卵巢大鼠骨生物力学和血清生化指标的影响

目的 研究抗氧化剂VitC、VitE及还原璎谷光甘肽(GSH)不同配伍用药方法 对去卵巢骨质疏松症大鼠骨生物力学及血清生化指标的影响,探讨抗氧化剂不同方案对去卵巢大鼠骨质疏松症的治疗作用.方法 4个月龄雌性SD大鼠70只,随机取50只行双侧卵巢切除术,20只行假手术.3个月后随机从手术组和假手术组各取10只大鼠检测体质量、子宫湿蕈、左侧股骨及腰椎骨密度、生物力学特性和血清生化指标Ca<'2+>、肌肝(Cr)、碱性磷酸酶(ALP)水平,以确定骨质疏松模型建立成功.确定模型建立成功后,其余动物分为A(假手术)、B(去卵巢生理盐水对照组,OVX control)、C(VitC+VitE)、D(GSH)、E(VitC+VitE+GSH)五组,每组10只.VitC[750 mg/(kg·d)]、GSH[125 mg/(kg·d)]腹腔注射,VitE[250 mg/(kg·d)]灌胃,模型组每天以生理盐水腹腔注射.3个月后,检测各组动物左侧股骨及第5腰椎生物力学特性和血清生化指标. 结果 检测骨质疏松模型实验中,模型组动物同假手术组相比,体质量明显增加,子宫萎缩,子宫湿重显著降低,左侧股骨及腰椎骨密度明显降低,左侧股骨生物力学最大载荷显著降低,血清Ca<'2+>、ALP、Cr水平升高.抗氧化剂治疗3个月后,与模型组相比,治疗组D、E组左侧股骨最大载荷、弹性载荷以及第5腰椎最大载荷均明显增加;血清ALP各组均显著降低;超氧化物歧化酶(SOD)、谷光苷肽过氧化物酶(GSH-Px)水平和血清抑制OH<'->能力D、E组显著升高;丙二醛水平C、D组显著降低;各组血清Ca<'2+>水平无明显改变. 结论 抗氧化刺崩药组合GSH和GSH+VitC+VitE能明显改善骨生物力学最大载荷和弹性载荷及血清抗氧化指标抑制OH<'->能力、SOD和GSH-Px,对骨质疏松具有较显著改善作用.     

墨旱莲对维甲酸所致大鼠骨质疏松症的药效学研究

目的探讨中药墨旱莲对维甲酸所致大鼠骨质疏松症的药效作用。方法 3月龄SPF级雌性SD大鼠60只,随机分为正常对照组、模型组(RA,75 mg/(kg·d))、墨旱莲组(1.46、0.73、0.37 g/(kg·d))、仙灵骨葆组(1.5 g/(kg·d))。除正常组外,其余各组给予维甲酸造模2 w,造模同时给予墨旱莲、仙灵骨葆。实验过程每日称量体重,连续给药6w后测定血钙(S-Ca)、血磷(S-P)、血清中碱性磷酸酶(ALP)和骨钙素(OCN)的水平,尿液中钙(U-Ca)、磷(U-P)、脱氧吡啶啉(DPD)的水平。采用DXA型骨密度仪检测大鼠的股骨、第4椎骨、胫骨的骨密度(bone mineral density,BMD)。三点弯曲试验检测左侧股骨生物力学性能:最大载荷、结构硬度、能量吸收、最大应力、弹性模量。Micro CT法分析右侧股骨骨微结构。结果墨旱莲1.46 g/(kg·d)能显著升高模型组大鼠的血钙水平,同时降低尿钙、ALP、OCN和DPD(P0.05)水平。与模型组相比,墨旱莲1.46 g/(kg·d)对维甲酸所致骨质疏松大鼠的股骨、第四椎骨及胫骨骨密度分别提高8.17%、11.79%、14.59%(P0.05),对最大载荷、结构硬度、能量吸收、最大应力、弹性模量等生物力学参数分别提高13.98%、16.33%、40.18%、12.45%、34.96%(P0.05),同时能有效抑制维甲酸所致大鼠股骨干骺端骨小梁微结构的退化(P0.05)。结论墨旱莲1.46 g/(kg·d)对维甲酸所致大鼠的骨质疏松症有防治作用,其作用机制可能与增强钙吸收、促进成骨细胞活性、降低骨转换率有关。     

姜黄素对APP/PS1转基因鼠骨微结构及骨密度的影响

目的应用Micro-CT定量研究用姜黄素治疗前后APP/PS1转基因鼠胫骨近端的骨微结构及骨密度的变化。方法 3月龄雌性小鼠(n=6/组)分为以下3组:野生型小鼠(对照组),APP/PS1转基因鼠(模型组)和APP/PS1转基因鼠+姜黄素(用药组)。对照组和模型组自由摄食和饮水;用药组,姜黄素按600mg·kg-1加入正常小鼠饲料,3组小鼠喂养至12月龄时处死,取其胫骨,应用Micro-CT进行扫描分析。结果 APP/PS1转基因组小鼠胫骨近端骨微结构、骨密度参数明显小于野生型小鼠(P0.05);APP/PS1转基因小鼠用药后胫骨近端骨微结构、骨密度参数明显大于用药前(P0.05),接近于野生型小鼠(P0.05)。结论姜黄素有助于提高APP/PS1转基因组小鼠的骨小梁数目、骨小梁厚度,骨密度,对APP/PS1转基因组小鼠骨微结构的改善有一定的治疗作用。     

自噬在镉致小鼠睾丸血-睾屏障损伤中的作用

目的:探究自噬在氯化镉致小鼠睾丸血-睾屏障(BTB)损伤中的作用。方法:将20只4周龄雄性C57BL/6小鼠随机分为对照组[0 mg/(kg·d)]、低剂量组[0.5 mg/(kg·d)]、中剂量组[1.0 mg/(kg·d)]和高剂量组[2.0 mg/(kg·d)],腹腔注射氯化镉，连续28 d。采用HE染色法分析睾丸组织形态学变化，生物示踪法观察BTB的完整性，Western印迹检测BTB组分ZO-1和N-Cadherin蛋白的表达。采用0、2.5、5和10μmol/L CdCl₂处理睾丸支持细胞株(TM4细胞)24 h, Western印迹检测ZO-1和N-Cadherin蛋白，以及自噬相关蛋白LC3II和p62的变化情况。在含CdCl₂(10μmol/L)的细胞培养液中分别加入自噬抑制剂氯喹(CQ)(5μmol/L)或自噬激动剂雷帕霉素(Rap)(50 nmol/L)处理细胞24 h, Western印迹检测LC3II、p62、ZO-1和N-Cadherin蛋白表达。结果:与对照组小鼠相比，镉染毒组小鼠的生精小管间隙增大、生精细胞内空...     

金雀异黄酮对去卵巢大鼠椎体力学性能的影响

目的 探讨皮下注射金雀异黄酮对OVX大鼠椎体力学性能的影响及骨力学性能的决定因素.方法 40只7月龄雌性SD大鼠随机分为去卵巢组(OVX组)、假手术组(SHAM)、OVX+17β雌二醇干预组(EST,10 μg/(kg·d))、OVX+金雀异黄酮干预组(GEN, 5 mg/(kg·d)).手术后15 w第5腰椎进行压缩试验,第6腰椎先行显微CT扫描测量骨密度和微结构参数,然后进行疲劳损伤试验,最后行大块组织品红染色、塑料包埋和磨片,磨片用于微损伤、骨细胞密度检测.结果 去卵巢后15 w,与SHAM组相比,OVX大鼠椎体骨密度、微结构参数、骨细胞密度和最大应力均降低(P<0.05),微破裂密度和面密度增加(P<0.05).GEN组骨小梁连接密度较OVX组增加(P<0.05),其他微结构参数与OVX组差异无统计学意义.与OVX组相比,GEN和EST替代组骨细胞密度和最大应力均增加(P<0.05),微破裂密度和面密度减少(P<0.05).OVX组、GEN和EST替代组之间骨密度和弹性模量差异无统计学意义.结论 金雀异黄酮对去卵巢大鼠椎体骨力学性能的维持不依赖BMD和微结构变化.     

Bone turnover biomarkers and risk of osteoporotic hip fracture in an Asian population

While epidemiologic studies suggest that bone turnover biomarkers may predict hip fracture risk, findings are inconsistent and Asian data are lacking. We conducted a matched case–control (1:1) study nested in the Singapore Chinese Health Study, a population-based prospective cohort of Chinese men and women (45–74 years) recruited from 1993 to 1998 in Singapore. One hundred cases with incident hip fracture and 100 individually matched controls were randomly selected from 63,257 participants. Serum bone turnover biomarkers, namely bone alkaline phosphatase (bone ALP), osteocalcin (OC), procollagen type I N propeptide (PINP), N-terminal and C-terminal crosslinking telopeptide of type I collagen (NTX-I and CTX-I) were measured using immunoassays. Hip fracture cases had significantly higher serum levels of OC, PINP, CTX-I and NTX-I than controls (p < 0.05). There was a dose-dependent positive relationship between OC, PINP, CTX-I and NTX-I and risk of hip fracture (all Ps for trend ≤ 0.006), where the risk was significantly increased by 4.32–8.23 folds for the respective BTM [Quartile (Q) 4 vs. Q1]. The odds ratio [OR (95% CI)] at the highest quartile (Q4) was 6.63 (2.02–21.18) for PINP and 4.92 (1.67–14.51) for CTX-I. The joint effect of PINP and CTX-I showed a 7-fold increase in risk (OR: 7.36; 95% CI: 2.53–21.41) comparing participants with higher levels of PINP (Q4) and CTX-I (Q3–Q4) to those with low levels of PINP (Q1–Q3) and CTX-I (Q1–Q2). Our data demonstrated that higher serum levels of bone turnover biomarkers were associated with increased risk of hip fracture in an Asian population.     

Prognostic value of PINP, bone alkaline phosphatase, CTX-I, and YKL-40 in patients with metastatic prostate carcinoma

BACKGROUND: To examine the prognostic value of markers of bone metabolism (serum PINP, BAP, and CTX-I) and serum YKL-40 in metastatic prostate carcinoma (PC). METHODS: The biomarkers were determined by ELISAs in 153 metastatic PC patients before treatment with parenteral estrogen or total androgen ablation. The median follow-up was 4.9 years. One hundred fifteen patients died. RESULTS: The biomarkers were increased in the patients compared to controls (P < 0.001), and related to performance status and Soloway score (except YKL-40), but not to T-category and WHO tumor grade. PINP was elevated in 87%, BAP (55%), CTX-I (33%), and YKL-40 (43%). Univariate analysis showed an association to survival: PINP (HR = 1.6, P < 0.0001), BAP (HR = 1.4, P < 0.0001), CTX-I (HR = 1.7, P < 0.0001), and YKL-40 (HR = 1.4, P = 0.004). In multivariate Cox analysis performance status, WHO grade, Soloway score, PINP, and YKL-40 were independently predictive factors. CONCLUSIONS: High serum PINP, BAP, CTX-I, and YKL-40 are associated with poor outcome of metastatic PC patients.     

High bone turnover is an independent predictor of mortality in the frail elderly.

Osteoporotic fractures are associated with accelerated bone turnover and excess mortality. In a prospective study of 1112 frail subjects (79% female; mean age, 86 years), high bone turnover was an independent predictor of all-cause mortality. This association seemed to be mainly manifested in deaths from cardiovascular causes. INTRODUCTION: Osteoporotic fractures are associated with accelerated bone turnover and excess mortality. In a prospective cohort study of elderly men and women, we assessed whether the rate of bone turnover measured by markers of bone remodeling is a direct predictor of mortality. MATERIALS AND METHODS: We measured serum concentrations of the aminoterminal propeptide of type I collagen (PINP), a marker of bone formation, and of the carboxyterminal telopeptide of type I collagen (CTX-I), a marker of bone resorption, along with serum PTH and 25-hydroxyvitamin D [25(OH)D] levels in 1112 subjects (79% female; mean age, 86 years) living in residential care. Co-morbidity was measured using the Implicit Illness Severity Scale. Fracture data were validated by a radiology report. Mortality and causes of death were ascertained from death certificates. RESULTS: Over a median follow-up of 817 days, 559 (50.3%) subjects died. In univariate analyses, time to death from all causes was significantly (p < 0.01) associated with age (HR = 1.62 per 10 years), male sex (HR = 1.33), immobility (HR = 1.94), co-morbidity (HR = 0.31, mild versus severe), lower weight (HR = 0.83 per 10-kg increase), impaired cognitive function (HR = 2.14, severe versus normal), number of medications (HR = 1.05 each), hip fracture (HR = 2.26), log serum creatinine (HR = 1.67), log PTH (HR = 1.29), CTX-I (HR = 1.70, highest 25% versus lowest 75%), and PINP (HR = 1.46, highest 25% versus lowest 75%). In multivariate analysis adjusting for age, sex, immobility, co-morbidity, weight, cognitive function, number of medications, PTH, and hip fracture status, the highest quartile was significantly more likely to die than the rest for both serum CTX-I (HR = 1.39; 95% CI: 1.14-1.70; p = 0.002) and PINP (HR = 1.25; 95% CI: 1.02-1.52; p = 0.03). For individual causes of death, CTX-I was significantly associated with deaths from cardiac causes (HR = 1.78: 95% CI: 1.27-2.50; p < 0.001). CONCLUSIONS: We conclude that in the frail elderly, high bone turnover is associated with all cause mortality independently of age, sex, health status, serum PTH levels, and hip fracture status. The mechanism of the effect of bone turnover on mortality seems to be mainly manifested in deaths from cardiovascular causes.     

依普拉芬对去卵巢大鼠骨质疏松骨保护效果及NO调控机制研究

目的 研究人工合成的植物雌激素-依普拉芬对去卵巢大鼠骨密度及生物力学的影响;同时与雌激素对照,探讨其骨保护效果及NO调控作用机制.方法 60只6月龄SD雌性大鼠,随机分成6组:假手术组和手术切除大鼠双侧卵巢组,后者分为阴性对照组、依普拉芬低、中、高剂量组和雌激素对照组, 分别给予基础饲料和不同剂量受试物,12周后进行骨密度、血清NO及NOS浓度测定;免疫组化方法检测股骨NOS表达.结果 与假手术组相比,去卵巢大鼠阴性对照组股骨密度、生物力学指标和血清NO、eNOS浓度以及股骨eNOS表达明显降低,血清iNOS浓度以及股骨iNOS表达增加,依普拉芬组骨密度、生物力学指标血清NO、eNOS浓度以及股骨eNOS表达均高于阴性对照组,与假手术组差异无显著意义.同时低于雌激素组.血清iNOS浓度以及股骨iNOS表达均低于阴性对照组,与假手术组以及雌激素组差异无显著意义.结论 一定浓度的依普拉芬可以通过提高去卵巢大鼠eNOS的表达来提高NO的浓度,促进成骨,增加骨密度,达到防治骨质疏松症的作用.     

辛伐他汀延缓生酮饮食导致的骨量丢失的实验观察

目的分析辛伐他汀(ST)治疗后小鼠股骨骨微结构、力学性能及骨形态的改变,评价ST对缓解生酮饮食引起的骨量丢失的治疗作用。方法 40只C57小鼠随机分为假手术组、卵巢切除组(OVX)、卵巢切除加辛伐他汀组(OVX+ST)、生酮饮食组(KD)及生酮饮食加辛伐他汀组(KD+ST),每组8只。OVX和OVX+ST组小鼠行双侧卵巢切除,予以常规标准饮食。KD及KD+ST组予以碳水化合物与脂肪比为1∶3的生酮饮食,OVX+ST及KD+ST组予以ST灌胃,剂量为20 mg/(kg·d)。12周处死取材,测量各组小鼠血酮、血糖、血钙、血磷等的水平,分别采用显微CT扫描、三点弯曲试验、有限元分析及HE染色观察股骨骨量微结构、力学性能及骨形态。结果 KD组和KD+ST组小鼠血酮含量为1. 28 mmol/L和1. 48 mmol/L,明显高于假手术组(0. 60 mmol/L)。各组间血糖、血钙或血磷的差异没有统计学意义。辛伐他汀治疗提高了KD+ST组的松质骨骨量、股骨有限元压缩刚度及三点弯曲刚度,其中KD+ST组较KD组股骨远端松质骨骨体积分数从4. 1%上升到6. 6%,有限元压缩刚度从35 N/mm提高到161 N/mm,股骨三点弯曲刚度从44 N/mm提高到60 N/mm,组织学观察也表明KD+ST组的股骨远端松质骨骨小梁较KD组明显增多。结论辛伐他汀治疗提高了KD组小鼠股骨松质骨骨体积分数、三点弯曲刚度、有限元压缩刚度和骨小梁数目,缓解了生酮饮食导致的骨量丢失。     

续苓健骨方联合阿仑膦酸钠调控EphB4/EphrinB2双向通路对去卵巢骨质疏松大鼠的研究

目的 观察续苓健骨方联合阿仑膦酸钠对去卵巢快速骨丢失模型大鼠骨密度、结构、骨代谢和EphB4/EphrinB2信号通路的影响,探讨其作用机制。方法 50只雌性SD大鼠随机分为5组：假手术组、模型组、续苓健骨方组、阿仑膦酸钠组和联合用药组,行双侧卵巢切除术制备模型;药物分别干预12周后取材,DXA检测股骨骨密度,HE染色观察胫骨组织显微结构,ELISA检测血清Ⅰ型胶原交联C末端肽(S-CTX)和Ⅰ型原胶原N端前肽(PINP)水平,Real-time PCR和Western blot检测腰椎EphB4、EphrinB2 mRNA和蛋白的表达水平。结果 与假手术组相比,模型组骨密度显著降低(P＜0.001),骨小梁稀疏并且排列紊乱,S-CTX和PINP含量均增高(P＜0.01),骨组织EphB4、EphrinB2 mRNA和蛋白表达水平均显著降低(P＜0.001)。与模型组相比,3个药物组骨密度均增高,其中以联合用药组最佳(P＜0.001),骨小梁较密并且排列规则,S-CTX含量不同程度降低,PINP含量不同程度增高,EphB4、EphrinB2 mRNA和蛋白表达水平均增高。结论 续苓健骨方联合阿仑膦酸钠起协同作用,可能通过激活EphB4/EphrinB2信号通路改善骨组织显微结构和血清骨代谢指标,从而提高大鼠骨密度起抗骨质疏松作用。     

肾性贫血动物模型的制作

目的:应用适量马兜铃酸(aristolochic acid,AA)探索创建肾性贫血小鼠模型。方法:6周龄雄性C57BL/6小鼠分3组,正常对照组;马兜铃酸腹腔注射组:AA/2d组(马兜铃酸3 mg·kg^-1·2 d^-1)和AA/3d组(3 mg·kg^-1·3 d^-1),共给药6周。造模第6、9、12周观察肾功能、贫血指标、肾脏病理评分、肾纤维化及肾组织EPO的蛋白表达情况。结果:给药6周时,血肌酐AA/2d组(29.9±1.5)μmol/L和AA/3d组(31.0±1.9)μmol/L均明显高于对照组(8.7±1.4)μmol/L,P<0.001;Hb水平AA/2d组(88.3±3.1)g/L、AA/3d组(85.7±7.4)g/L都明显低于对照组(148.7±4.9)g/L,P<0.001;HCT值AA/2d组、AA/3d组分别为0.30±0.01、0.29±0.02还是都明显低于对照组(0.51±0.02),P<0.001;肾脏病理呈现明显的肾间质纤维化,肾组织中EPO蛋白表达水平下降,提示肾性贫血模型成功。连续观察9、12周,上述变化持续存在,但AA/2d、AA/3d组间的临床病理指标无统计学意义。结论:3 mg·kg^-1·3 d^-1马兜铃酸连续给药6周可成功建立合格稳定的肾性贫血小鼠模型,该模型临床表现为不可逆的肾衰竭和贫血,肾组织中的促红素表达下降,肾脏病理表现为慢性肾间质纤维化。     

ApoE gene deficiency enhances the reduction of bone formation induced by a high-fat diet through the stimulation of p53-mediated apoptosis in osteoblastic cells.

Osteoblast apoptosis increased in the tibias of apoE(-/-) mice fed with a high-fat diet, decreasing bone formation. The expression of p53 mRNA in marrow adherent cells increased. LDL or oxidized LDL increased apoptosis in the calvarial cells of apoE(-/-) mice. The increase in p53-mediated apoptosis is apparently related to a high-fat diet-induced osteopenia in apoE(-/-) mice. INTRODUCTION: The effects of high-fat loading and the apolipoprotein E (apoE) gene on bones have not been elucidated. We hypothesized that apoE gene deficiency (apoE(-/-)) modulates the effects of high-fat loading on bones. MATERIALS AND METHODS: We assessed this hypothesis using wildtype (WT) and apoE(-/-) mice fed a standard (WTS and ApoES groups) or a high-fat diet (WTHf and ApoEHf groups). The concentration of serum lipid levels and bone chemical markers were measured. Histomorphometry of the femurs was performed using microCT and a microscope. Bone marrow adherent cells from the femurs were used for colony-forming unit (CFU)-fibroblastic (CFU-f) assay and mRNA expressions analysis. The apoptotic cells in the tibias were counted. TUNEL fluorescein assay and Western analysis were performed in cultures of calvarial cells by the addition of low-density lipoprotein (LDL) or oxidized LDL. RESULTS: In the ApoEHf group, the values of cortical bone volume and trabecular and endocortical bone formation of the femurs decreased, and urinary deoxypyridinoline increased. Subsequent analysis revealed that the number of apoptotic cells in the tibias of the ApoES group increased, and more so in the ApoEHf group. The ratio of alkaline phosphatase-positive CFU-f to total CFU-f was decreased in the ApoEHf group. p53 mRNA expression in adherent cells of the apoE(-/-) mice increased and had a significantly strong positive correlation with serum LDL. TUNEL fluorescein assay of osteoblastic cells revealed an increase of apoptotic cells in the apoE(-/-) mice. The number of apoptotic cells in the apoE(-/-) mice increased with the addition of 100 microg/ml LDL or oxidized LDL. The p53 protein expression in apoE(-/-) cells exposed to 100 microg/ml LDL or oxidized LDL increased. CONCLUSIONS: We concluded that apoE gene deficiency enhances the reduction of bone formation induced by a high-fat diet through the stimulation of p53-mediated apoptosis in osteoblastic cells.     

朝藿定C联合骨髓间充质干细胞移植对糖皮质激素性骨质疏松小鼠骨密度和骨代谢的影响

目的 探讨朝藿定C联合骨髓间充质干细胞移植对糖皮质激素性骨质疏松小鼠骨密度和骨代谢的影响。方法 75只雄性C57BL/6小鼠分为空白对照组、模型组、朝藿定C组、干细胞移植组、联合组，每组各15只。空白对照组以0.1 mL生理盐水肌肉注射，其余4组以0.1mL地塞米松肌肉注射，2次/周，连续干预8周，以骨密度值确定造模成功。造模成功后7 d分别进行骨髓间充质干细胞移植和朝藿定C灌胃治疗，连续8周。测定骨密度和骨结构参数、血清骨代谢指标水平以及AKT蛋白磷酸化（p-AKT）水平。结果 与空白对照组比较，模型组体质量、骨矿物质含量（BMC）、骨矿物质密度（BMD）、组织矿物质含量（TMC）、组织矿物质密度（TMD）、骨体积分数（BV/TV）、骨小梁数量（Tb.N）、骨小梁厚度（Tb.Th）以及骨源性碱性磷酸酶（BALP）、Ⅰ型前胶原羧基端前肽（PINP）、AKT蛋白磷酸化水平显著降低，骨小梁分离度（Tb.Sp）显著增加，血清骨钙素（OCN）和抗酒石酸酸性磷酸酶（Trap）水平显著升高（P＜0.05）。与模型组比较，朝藿定C组、干细胞移植组和联合组小鼠体质量、BMC、BMD、TMC、TMD、BV/TV、Tb.N、Tb.Th显著增加，BALP、PINP、AKT蛋白磷酸化水平显著升高，Tb.Sp、血清OCN和Trap水平显著降低，且联合组上述指标改善优于其余2组（P＜0.05）。结论 朝藿定C联合骨髓间充质干细胞移植可促进糖皮质激素性骨质疏松小鼠骨形成，增加骨密度，改善骨微结构，其机制可能与提高骨代谢水平和活化PI3K/AKT通路有关。