重组人生长激素治疗大鼠卵巢切除后骨量丢失的研究

目的研究重组人生长激素对卵巢切除后大鼠骨量丢失的治疗作用。方法大鼠卵巢切除后３个月开始接受不同剂量生长激素治疗,８周后处死。采用骨密度测定,骨组织计量学分析,血清生化指标测定等观察生长激素对实验性骨质疏松症的治疗作用。结果生长激素使骨质疏松大鼠腰椎骨增加,股骨中段皮质骨显著增厚,腰椎体小梁骨体积增加而且小梁连结性明显改善。重组人生长激素可促进骨质疏松大鼠类胰岛素生长因子－１合成增加。结论重组人生长     

联合运用重组人生长激素和雌激素对去势大鼠骨密度及生物力学的影响

目的研究重组人生长激素和雌激素联合用药对去势大鼠股骨、下颌骨骨密度及生物力学的影响。方法选4月龄SD雌性大鼠,随机分为5组:假手术组(sham-OVX)、骨质疏松组(OVX)、雌激素组(E2)、重组人生长激素组(rh-GH)、联合用药组(E2 rh-GH),于去势后8w,各组分别腹腔给予相应的药物,4w末及8w末分两批处死各组大鼠,取股骨和下颌骨新鲜标本,进行骨密度(BMD)与最大载荷的实验研究。结果用药4w末及8w末,OVX组下颌骨和股骨BMD均显著低于其他4组。E2 rh-GH组下颌骨BMD较E2组和rh-GH组高,但是差异均无统计学意义(P>0.05)。用药4w末及8w末,OVX组下颌骨和股骨最大载荷均显著低于sham-OVX组和用药组。E2 rh-GH组下颌骨及股骨最大载荷比E2组、rh-GH组高,用药4w末联合用药组与单一用药组最大载荷的差异有统计学意义;而用药后8w末差异均无统计学意义。结论雌激素、重组人生长激素及两者联合用药都能有效的抑制骨质疏松大鼠颌骨及股骨骨量的丢失,提高骨强度。联合用药与单一用药对下颌骨骨密度的影响没有明显的差异,可能与测量的方法和测量部位结构有关。联合用药对股骨骨密度、最大载荷及颌骨最大载荷的作用与用药时间有关,短期内效果优于单独用药。     

大鼠体外循环后肝损害与生长激素及其结合蛋白变化的关系及意义

肝脏是生长激素(GH)/胰岛素样生长因子-I(IGF-I)轴的中心器官,体外循环(CPB)时是否存在GH分泌不足及GH抵抗报道尚少。我们通过建立大鼠CPB模型,观察CPB围术期肝损害与GH-IGF-I轴的变化的关系。     

补充雌激素对卵巢切除大鼠血清皮质醇和IGF-1的影响

目的研究成年雌性SD大鼠于卵巢切除后及再补充雌激素后血清皮质醇和类胰岛素生长因子-Ⅰ(IGF-Ⅰ)的变化。方法成年雌性SD大鼠分为假手术对照组、去势组、去势 补充雌激素组。术后5个月处死,采取血清检测雌二醇、皮质醇及IGF-Ⅰ的水平。结果 大鼠卵巢切除后雌激素水平显著降低(P<0.01),而皮质醇水平明显升高(P<0.05),IGF-Ⅰ无明显变化;应用雌激素治疗,可以显著降低血清皮质醇(P<0.01)和IGF-Ⅰ(P<0.01)的水平。结论 卵巢切除及补充雌激素对大鼠血清皮质醇和IGF-Ⅰ有显著影响。     

氨基酸溶液和重组人生长激素对肝硬化大鼠生长激素-胰岛素样生长因子1轴的影响

目的探讨氨基酸溶液和重组人生长激素(rhGH)对肝硬化大鼠生长激素(GH)-胰岛素样生长因子1(IGF—1)轴的影响。方法肝硬化大鼠随机分为术前组、肝部分切除后1d组、术后肠外营养用复方氨基酸注射液(Novamin)5d组、用10％复方氨基酸注射液(Hepa)5d组、用rhGH Hepa 5d组,各组均为6只大鼠,对照组为正常大鼠。分别测大鼠肝功能、血糖及血清GH、IGF-1、胰岛素样生长因子结合蛋白3(IGFBP-3)水平,用逆转录聚合酶链反应检测肝白蛋白(ALB) mRNA、IGF-1 mRNA、IGFBP-3m RNA的表达,肝组织行Ki67免疫组化染色。结果用Hepa的肠外营养组5d后与用Novamin组比较,GH水平明显降低,肝ALB mRNA、IGF-1m RNA、IGFBP-3 mRNA表达水平明显升高,肝Ki67指数无差别;而rhGH Hepa组肠外营养5d后比Hepa组血糖、GH、IGF-1、IGFBP-3水平明显升高,肝ALB mRNA、IGF-1 mRNA表达水平及肝Ki67指数也明显升高。结论氨基酸溶液和重组人生长激素均可影响肝硬化大鼠生长激素．胰岛素样生长因子1轴,检测血清IGF-1、IGFBP-3水平有助于营养素的选择和评估。     

雌激素对去卵巢大鼠松质骨中骨保护素mRNA表达的影响

目的 :通过去卵巢大鼠予雌激素后 ,腰椎内OPGmRNA的表达进一步探讨雌激素防治骨质疏松的分子机制。方法 :雌性 3月龄SD大鼠 3 6只 ,随机均分为卵巢切除 +雌激素 (倍美力 )预防组 (EP) ,骨质疏松组 (卵巢切除OVX)及正常对照组 (SHAM ) 3组。术后 12周处死大鼠 ,取第 3腰椎椎体 ,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OPGmRNA的表达。同时取第 4腰椎椎体 ,行骨组织形态计量学检测。结果 12例腰椎骨组织中 ,EP组 10例OPGmRNA呈阳性表达 (阳性率 83 .3 % ) ,OVX组 2例呈阳性表达 (阳性率 16.7% ) ,而SHAM组 12例均呈阳性表达。统计学处理 :EP组与OVX组有高度显著性差异 (P <0 .0 1) ,EP组与SHAM组无显著性差异 (P >0 .0 5 )。骨组织形态计量学检测 ,OVX组骨体积、骨小梁数目均比SHAM组显著减少 ,而骨小梁间隔显著增大。与OVX组比较 ,EP组骨体积、骨小梁厚度增加 ,骨小梁间隔稍有减小 ,无统计学意义。EP对骨小梁数目无明显影响。结论 :雌激素通过增强松质骨中OPGmRNA的表达 ,抑制破骨细胞性骨吸收 ,从而对去卵巢大鼠的骨质疏松有明显的预防作用。     

补骨胶囊与17α-炔雌醇联合用药对大鼠骨质疏松的预防及对子宫刺激作用的研究

目的 观察补骨胶囊与17α-炔雌醇联合用药对去卵巢大鼠骨质疏松的预防作用及对子宫的刺激作用。方法 47只4月龄SD雌性大鼠随机分为6组：①假手术组；②去卵巢组；③高剂量及④低剂量17α-炔雌醇预防组；⑤补骨胶囊预防组；⑥补骨胶囊与低剂量17α-炔雌醇联合用药组。实验12周。通过骨组织形态计量学方法分析大鼠骨量变化情况，通过测量子宫湿重及内膜厚度分析用药对子宫的刺激作用。结果 去卵巢组骨小梁面积百分率(Tb．Ar％)比假手术组显著下降，高剂量17α-炔雌醇组和联合用药组Tb．Ar％比去卵巢组显著增加。高、低剂量17α-炔雌醇组与去卵巢组比，子宫湿重及内膜厚度都显著增加；联合用药组与两个17α-炔雌醇组比，子宫湿重及内膜厚度显著降低。结论补骨胶囊与17α-炔雌醇联合用药可有效预防去卵巢大鼠骨质疏松，与单独使用17α-炔雌醇比，对子宫刺激作用显著降低。     

巴戟天含药血清对原代破骨细胞RANK和CA II mRNA表达的影响

目的 观察巴戟天含药血清对原代破骨细胞RANK和CA II mRNA表达的影响.方法 取SPF级大鼠48只,随机分成正常组12只和去势组36只,正常组切去卵巢周围部分脂肪,去势组切除卵巢.3个月后测定两组雌激素水平并将去势组随机分为骨质疏松组,骨质疏松+雌激素组,骨质疏松+巴戟天含药血清组.采用机械分离法提取各组的破骨细胞,培养5 d后,TRAP染色及电镜扫描骨片等的方法鉴定破骨细胞.最后用雌激素或巴戟天含药血清干预3 d,以RT-PCR法检测各组RANK和CAIImRNA表达.采用单因素方差分析或多样本均数两两比较进行统计分析.结果 去势组后大鼠雌激素水平低于正常组(P＜0.01).骨质疏松组破骨细胞RANK和CAII表达均高于正常组(P＜0.05,P＜0.05);巴戟天含药血清可降低骨质疏松后大鼠破骨细胞RANK和CA II的表达(P＜0.05,P＜0.05).结论 巴戟天和雌激素均可降低骨质疏松大鼠破骨细胞RANK和CAII的表达,从而达到抑制骨质疏松的作用.     

补肾方药对骨质疏松防治的实验研究

目的 探讨补肾方药对骨质疏松的防治作用及其机理。方法 本研究选用3种模型，即地塞米松和卵巢切除所诱发的骨质疏松模型，自然衰老的大鼠骨质疏松模型，并给予中药治疗。测定大鼠骨密度、骨代谢相关生化指标及骨形态计量学指标及基因表达分析。结果 补肾方药可显提高地塞米松所致骨质疏松大鼠的骨密度、血清骨钙素水平，降低尿钙排泄，同时，促进骨组织中I型胶原和LMP－1 mRNA表达，补肾方药还提高小肠黏膜CaBp-D9K基因表达，从而使小肠黏膜钙结合蛋白（CaBp)的合成增加，促进小肠对钙的吸收；补肾方药可提高卵巢切除所诱发的骨质疏松大鼠的骨密度，促进骨组织中I型胶原mRNA及小肠黏膜中VDR mRNA的表达，提高卵巢切除大鼠血清雌激素的水平，同时，提高骨组织中雌激素受体α和β（ERα，ERβ）的表达，增加雌激素对骨代谢的调节作用；补肾方药改善老年骨质疏松大鼠骨代谢，降低尿钙排泄，通过提高骨转换，主要是促进骨形成防止骨量丢失。结论 补肾方药通过多环节、多途径，调节骨质疏松大鼠的骨形成与骨吸收，使其达到骨形成与骨吸收相偶联，而防治骨质疏松。     

金匮肾气丸联合葡萄糖酸钙对去势大鼠骨质疏松的影响

目的观察金匮肾气丸联合葡萄糖酸钙对去卵巢大鼠骨质疏松的影响,探究其防治骨质疏松的机制。方法将75只雌性大鼠随机分成空白对照组,模型组,葡萄糖酸钙组,金匮肾气丸组,金匮肾气丸及葡萄糖酸钙联合用药组共5组,采用卵巢切除的方法制成骨质疏松动物模型,灌胃给药3个月后进行血钙,磷,骨钙含量的测定和组织病理学的观察。结果模型组与空白对照组相比,去势大鼠血清钙磷含量无明显变化,钙片组、金匮肾气丸组及联合组血钙,磷含量相比空白组也无明显差异;去势大鼠骨钙含量较正常大鼠明显降低,钙片组,金匮肾气丸,以及联合组去势大鼠骨钙含量较模型组明显改善,以联合组改善最明显(P<0.01)。组织病理学观察见:经钙片和金匮肾气丸治疗后骨皮质可见蚕食样改变区域变小,骨小梁较细,排列较为一致。联合用药组骨皮质厚度基本正常,骨小梁略变细,排列较整齐,骨小梁厚度基本正常。结论金匮肾气丸联合葡萄糖酸钙可以减少去卵巢骨质疏松大鼠骨矿物质的丢失,对骨质疏松具有较好的防治作用。     

荧光标记拔牙创口骨愈合过程中矿化沉积速度的研究

目的研究骨质疏松及雌激素替代治疗对拔牙创骨愈合的影响。方法选用12周龄雌性SD大鼠,随机分为假手术对照组和实验组,实验组行双侧卵巢切除术,对照组行假去势手术。术后8周将实验组随机分为骨质疏松组（OP）和雌激素治疗组,同时拔除所有大鼠左侧上颌第1磨牙。磨牙拔除术后7、30、60天分3批处死大鼠。上颌拔牙创口处制作不脱钙切片,行组织学及荧光观察。结果拔牙后7天时,3组荧光表现基本相似;拔牙后30天及60天骨质疏松组拔牙创骨矿沉积率明显低于假手术对照组及雌激素治疗组,雌激素治疗组骨矿沉积率低于假手术对照组。结论OP对拔牙后早期的新骨形成骨的矿化影响不大,但在拔牙创愈合中晚期可明显减少拔牙创新骨矿化。雌激素应用可明显减缓OP对拔牙创内新骨骨矿化的影响。     

Effect of intermittent administration of human parathyroid hormone (1-34) on the mandibular condyle of ovariectomized rats

Intermittent administration of human parathyroid hormone (1-34) (PTH) increases bone mass in lumbar vertebrae and long bones of osteoporotic experimental animals. However, whether PTH has the same effect on jaw bones remains unclear. This study determined the effect of intermittent administration of PTH on rat mandibular condyle affected by estrogen deficiency. Fifty 6-month-old rats were either sham operated or ovariectomized, then divided into five groups depending on surgical procedure and hormone administration: sham + vehicle (SV), OVX + vehicle (OV), OVX + PTH 6 μg/kg once per week (OP6-1), OVX + PTH 60 μg/kg once per week (OP60-1), and OVX + PTH 20 μg/kg three times per week (OP20-3). PTH or vehicle was injected intermittently for 6 months in 5 rats of each group either immediately after surgery in a preventive administration experiment, or injected starting 6 months after surgery in a therapeutic administration experiment. The mandibles were excised, and bone morphometry was performed using confocal laser scanning microscopy and soft X-ray images. In both experiments, the bone volume of the OV groups was significantly lower than that of the SV group (P < 0.01); also, depending on dose and frequency, the bone volume of the OP group was higher than that of the OV group, particularly in the OP20-3 group. The value of mineralized surface of the OP groups was significantly higher than that of the OV group (P < 0.01), whereas the value of eroded surface of the OP groups was not significantly higher than that of the OV group. This study indicates that preventive and therapeutic intermittent administration of PTH in ovariectomized rats increase the bone formation in rat mandibular condyle without accelerating bone resorptive activity. This anabolic effect was best induced by the injection mode of 20 μg/kg three times per week. Received: Dec. 21, 1998 / Accepted: March 24, 1999     

大豆苷元干预Hedgehog信号通路调控去卵巢骨质大鼠基质金属蛋白酶代谢

目的 探究大豆苷元对骨质疏松（osteoporosis,OP）模型大鼠Hedgehog信号通路的调控作用,分析大豆苷元对OP大鼠基质金属蛋白酶代谢的影响。方法 随机将60只雌性SPF级Wistar大鼠分为假手术组、模型组、大豆苷元低、中、高剂量治疗组5组,每组12只。除假手术组,其他4组均采用去卵巢法建立OP模型。造模成功后开始干预治疗,假手术组、模型组等剂量生理盐水灌胃,其他给予大豆苷元低、中、高剂量治疗,连续干预12周。酶联免疫吸附法测定各组大鼠血清雌激素（E2）、骨钙素（BGP）、血清碱性磷酸酶（ALP）含量水平;免疫组化法检测各组大鼠股骨组织中基质金属蛋白酶组织抑制剂-1（TIMP-1）和基质金属蛋白酶-7（MMP-7）免疫蛋白的表达情况;TUNEL染色法检测各组骨细胞凋亡情况;RT-PCR法和WB法检测各组骨组织中Shh、PTC1、Gli1 mRNA及蛋白表达水平。结果 大豆苷元能显著提高OP大鼠大鼠E2、TIMP-1水平,降低细胞凋亡及 BGP、ALP、MMP-7水平;显著改善股骨组织中Shh、PTC1、Gli1 mRNA及蛋白表达水平（P＜0.05）,且大豆苷元不同组具有剂量依赖性（P＜0.01）。结论 大豆苷元可有效调控OP模型大鼠基质金属蛋白酶代谢水平,这可能与大豆苷元能够有效调节Hedgehog信号通路表达相关。     

Insulinlike growth factor I gene expression is increased in the fetal lung after tracheal ligation

Background/Purpose

The mortality and morbidity in congenital diaphragmatic hernia are mainly caused by pulmonary hypoplasia. To improve clinical results, further methods inducing lung growth may have to be used. The aim of this report was to evaluate the expression of insulinlike growth factor I (IGF-I), estrogen receptor α, estrogen receptor β, growth hormone receptor, and thioredoxin in a rat model of hypoplastic, hyperplastic, and normal fetal lungs to improve understanding of lung growth.

Methods

Hypoplastic diaphragmatic hernia lungs were created by giving nitrofen by gavage to pregnant rats on day 9.5. Hyperplastic lungs were achieved by intrauterine tracheal ligation of rat fetuses on day 19. All lungs were harvested on gestational day 21. Total nucleic acids were extracted by proteinase K digestion and extraction in phenol/chloroform. The total nucleic acids mixture was hybridized with radioactively labeled RNA probes, and the radioactivity of the hybrids was compared with the respective standard curve of known amounts of in vitro synthesized mRNA. Immunohistochemistry staining was performed for IGF-I.

Results

The IGF-I mRNA was significantly (P < .01) higher in hyperplastic lungs compared with control and hypoplastic lungs. The latter 2 did not differ. No difference was found between the other mRNA levels in the study groups.

Conclusions

IGF-I is involved in the accelerated lung growth seen after intrauterine tracheal ligation.     

Combined estrogen and ghrelin administration decreases expression of p27<Superscript>kip1</Superscript> and proportion of isomyosin type I in the striated urethral and anal sphincters and levator ani of old ovariectomized rats

We compared estrogen and/or ghrelin effects on pelvic floor muscles in old versus young adult ovariectomized rats. Ovariectomized Fisher 344 rats (18 and 3 months old, n = 24 x 2) received 42 daily intraperitoneal 17-beta estradiol (10 mug kg(-1)), ghrelin (2 mug kg(-1)), both, or vehicle (n = 6 x 4/group). Cytoplasmic p27(kip1) expression and isomyosin I proportion in striated urethral and anal sphincters and levator ani were measured, respectively, by Western blot analysis and gel electrophoresis with immunohistochemistry of muscle ghrelin receptors and radioimmunoassay of circulating growth hormone. In young adult rats, estrogen significantly decreased cytoplasmic p27(kip1) and isomyosin I signal intensities. In old rats, ghrelin and estrogen/ghrelin significantly decreased both intensities with greater estrogen/ghrelin effect. Ghrelin receptors were not immunostained in any muscle. Estrogen and/or ghrelin significantly increased or decreased, respectively, circulating growth hormone in old and young adult rats. Estrogen/ghrelin administration reversed pelvic floor muscle ageing changes in old ovariectomized rats through growth hormone production.     

The effect of ovariectomy and ovarian steroid treatment on growth hormone and insulin-like growth factor-I levels in the rat femur.

Growth hormone (GH) and insulin-like growth factor-I (IGF-I) are known to play an important role in bone metabolism. The regulation of plasma levels of GH and IGF-I by ovarian steroids is well known, however, their effect on local GH and IGF-I is still unclear. In this study, we investigated the effect of ovariectomy and ovarian steroid treatment on the femur GH and IGF-I levels as well as on bone density in the rat. Nine month-old rats were ovariectomized (OVX) or sham-operated (SHAM) and 9 weeks after the surgery they were treated with daily s.c. injections of either 17beta-estradiol (OVX + E), progesterone (OVX + P), or vehicle (OVX + V) for another 10 weeks. GH and IGF-I levels in the femur extracts were measured by specific radioimmunoassay (RIA). Ovariectomy decreased GH and had no effect on IGF-I levels. Estradiol treatment increased femur GH and IGF-I levels compared to SHAM rats. Progesterone restored GH and increased IGF-I levels. Ovariectomy decreased, estrogen restored and progesterone partially restored femur bone density. Our results demonstrate that ovariectomy and ovarian steroids modulate the levels of GH and IGF-I in the bone of aged OVX rats. However, these effects appear to be limited to supraphysiological concentrations of estradiol and progesterone.     

抗疏健骨颗粒对去卵巢骨质疏松大鼠血清骨代谢及骨组织自噬水平的影响

目的研究抗疏健骨颗粒对去卵巢骨质疏松(osteoporosis,OP)大鼠骨稳态及自噬相关因子的影响,探讨抗疏健骨颗粒干预OP的作用机制。方法将60只SD大鼠随机分为5组:假手术组、模型组、抗疏健骨颗粒低、中、高剂量组。除假手术组外,其余4组建立去卵巢大鼠OP模型。正常喂养12周检测模型建成后,给予抗疏健骨颗粒干预12周,采用ELLSA法测定血清ALP、BGP和TRACP水平和股骨GPR48、ATF4含量;蛋白印迹法(Western blotting)检测股骨中AMPK、LC3、Beclin1和P62蛋白表达。结果抗疏健骨颗粒可显著降低OP大鼠血清ALP、BGP和TRACP水平,升高股骨GPR48、ATF4含量,上调AMPK磷酸化水平及LC3、Beclin1蛋白表达,降低P62水平,差异有统计学意义(P0.05)。抗疏健骨颗粒各剂量组中,中剂量组效果显著,与其他两组相比,差异有统计学意义(P0.05)。结论抗疏健骨颗粒可调节血清、骨组织中骨代谢指标,减轻去卵巢大鼠OP的发生,其机制可能与通过骨组织中AMPK调节自噬通路有关,通过恢复自噬水平,促进骨生成,进而维持骨平衡。