Dietary soy protein and isoflavones: minimal beneficial effects on bone and no effect on the reproductive tract of sexually mature ovariectomized Sprague-Dawley rats

OBJECTIVE: The present study was conducted to determine the effects of dietary soy protein and isoflavones on bone and the reproductive tract in the absence of the ovary. DESIGN: Three-month-old Sprague-Dawley rats (n = 56) were either sham-operated or ovariectomized and then fed diets containing casein or soy protein +/- isoflavone extract for 12 weeks. The amounts of casein, soy protein, and extract (per kg diet) in each group were as follows: (1) Ovariectomy, 200 g of casein; (2) Ovariectomy+low soy, 100 g of casein + 100 g of soy protein; (3) Ovariectomy+high soy, 200 g of soy protein; (4) Ovariectomy+low extract, 200 g of casein + 17.2 g of extract; (5) Ovariectomy+high extract, 200 g of casein + 34.4 g of extract; (6) Ovary intact, 200 g of casein; (7) Ovariectomy+estradiol-17beta, 200 g of casein. Diet consumption, body weight, uterine weight, urine deoxypyridinoline, and bone mineral density of the femur and lumbar vertebrae were measured. The femur rigidity was evaluated by histomorphometry. The reproductive tract (uterus, vagina, and cervix) was studied histologically. RESULTS: The Ovariectomy group showed significant increases in body weight, diet consumption, and deoxypyridinoline, decreases in uterine weight and bone mineral density, and negative changes in histomorphometry compared with the Ovary intact group. Neither soy protein nor extract diets abrogated these alterations, except for the Ovariectomy+high extract group that showed statistically significant positive changes in histomorphometric parameters. There were no histological differences in the reproductive tract among Ovariectomy, Ovariectomy+soy, and Ovariectomy+extract groups. The estradiol-17beta replacement abrogated ovariectomy-induced alterations. CONCLUSION: Dietary intake of isoflavones by sexually mature ovariectomized rats has a minimal beneficial effect on bone with no effect on the reproductive tract.     

Combination of soy protein and high dietary calcium on bone biomechanics and bone mineral density in ovariectomized rats

OBJECTIVE: To determine if feeding soy in combination with a high-calcium diet would preserve bone mineral density and biomechanical bone strength to a greater extent than either soy or calcium alone. DESIGN: Rats were sham-operated (n = l0) and fed a control diet (AIN93G containing 0.2% calcium, 20% casein) or ovariectomized (n = 40) and randomized to one of the following diets (n = 10 per group): control, high calcium (2.5% calcium, 20% casein), soy (0.2% calcium, 20% soy protein), or soy plus high calcium (2.5% calcium, 20% soy protein) for 8 weeks. Bone mineral density of femurs and lumbar vertebrae 1 through 6 were measured by dual energy x-ray absorptiometry. Biomechanical strength properties of femurs and the fifth lumbar vertebrae were measured by three-point bending and compression, respectively. RESULTS: The dietary combination of soy and high calcium did not result in higher femur bone mineral density compared with other ovariectomized groups, and there were no differences in femur yield load or peak load among groups. In contrast, soy plus high calcium resulted in a higher (P < 0.05) vertebral bone mineral density compared with all other ovariectomized groups. Vertebral strength was preserved among rats fed either soy plus high calcium, soy, or high calcium whereas the ovariectomized group fed a control diet had lower (P < 0.05) vertebral strength than the sham-operated group. CONCLUSION: Bone mineral density of the lumbar spine was the only bone outcome that significantly benefited from the combination of soy and high calcium compared with soy or high calcium alone.     

葛根异黄酮联合维生素D防治骨质疏松的协同效应

背景:葛根异黄酮具有类雌二醇结构特征,对去卵巢大鼠的骨质丢失和骨量的减少有一定的抑制作用。但也有报道其对去卵巢大鼠骨质疏松的防治效果较弱且单独使用效果不明显。 目的：观察葛根异黄酮与维生素D联合对去卵巢大鼠骨质疏松的作用。 方法：将81只3月龄雌性SD大鼠随机分为9组: 除假手术组外其他各组均行双侧卵巢摘除造模。低、中、高剂量TIP组,VitD组,低、中、高剂量联合组在去卵巢基础上分别灌胃给予相应剂量的葛根异黄酮和（或）维生素D;模型组和假手术组不给予药物治疗。给药3个月,计算大鼠子宫系数;测定大鼠血清碱性磷酸酶、血钙、血磷、骨钙素及血清雌二醇水平;检测大鼠股骨骨密度。 结果与结论：与模型组比较,中、高剂量TIP组,低、中、高剂量联合组大鼠子宫系数、雌二醇水平明显增高(P < 0.05),血清碱性磷酸酶、骨钙素水平明显降低(P < 0.05),股骨中心及远心骨密度均明显增加(P < 0.05),均以高剂量联合组作用效果最明显。且在子宫系数,雌二醇、血清碱性磷酸酶、骨钙素水平及股骨远心端骨密度方面,葛根异黄酮和维生素D均表现为协同作用。在血钙、血磷方面,葛根异黄酮和(或)维生素D的作用不明显。说明葛根异黄酮和维生素D联合使用对去卵巢大鼠骨质疏松的防治有协同效应。     

Isoflavone-supplemented soy yoghurt associated with resistive physical exercise increase bone mineral density of ovariectomized rats

OBJECTIVE: To determine the effects produced by the ingestion of a fermented soy product (soy yoghurt), supplemented with isoflavones and associated with a resistive exercise program, on the bone metabolism of mature ovariectomized (Ovx) and sham-ovariectomized (sham-Ovx) rats. METHODS: A total of 56 rats were used. They were divided into 2 sedentary control groups, the Ovx control group (C-Ovx) and the sham-Ovx control group (C-Sovx), each with 7 sedentary animals, and 2 treated groups, Ovx and sham-Ovx, with 21 animals each. These two treated groups were subdivided into three subgroups of seven animals each, which received the following treatments: consuming the soy yoghurt+sedentary, only subjected to resistive exercise, and consuming the soy yoghurt+resistive exercise. Both the program of resistive exercise and the consumption of soy yoghurt (at 3 mL/(kg body weight day)) continued for 12 weeks. The soy yoghurt was supplemented with isoflavones at 50mg/100g of product. The animals were sacrificed and their right-side femurs and tibias removed and assessed for bone mineral density (BMD). The alkaline phosphatase activity (AP) was determined in the blood serum. RESULTS: There was a significant increase in both femur and tibia BMD values and in serum alkaline phosphatase activity in all the treated subgroups, compared with the control groups (p<0.05). CONCLUSION: The ingestion of the soy yoghurt supplemented with isoflavones was capable of preventing a loss of bone mass in Ovx rats and of increasing bone mass in sham rats, whilst the resistive exercise program was effective in augmenting the bone mass in sham and Ovx rats.     

Characterization of the pharmacologic profile of a standardized soy extract in the ovariectomized rat model of menopause: effects on bone, uterus, and lipid profile

OBJECTIVE: This study was aimed to assess the effect of a standardized soy extract (SSE, Soyselect) in the ovariectomized rat model of menopause. DESIGN: Ovariectomized rats were treated for 6 weeks with the soy extract (50 or 100 mg/kg/day - PO), vehicle (distilled water), or 17beta-estradiol (0.5 mg/kg/day - PO). Tissue-specific estrogen agonist effects were examined using the endpoints bone mineral density, biochemical parameters of bone turnover, modulation of cytokines involved in the bone remodeling, uterine weight, uterine histology, uterine hormone receptor status, and serum lipid level. RESULTS: The SSE produced a bone-sparing effect associated with a slowing down in the increased bone turnover observed after ovariectomy (as indicated by measurements of serum osteocalcin levels and excretion ratio of deoxypyridinoline); changes in serum interleukin-6 levels observed after SSE suggested that this bone-sparing effect could be partly attributed to the modulation of osteoclastogenesis induced by interleukin-6. Remarkably, organ weight data and histopathologic analysis did not show any stimulatory activity of the SSE on the uterus. Immunohistochemical analysis showed a significant down-regulation of estrogen receptor-alpha (ERalpha) in uterine epithelium after 17beta-estradiol treatment, but not after treatment with the SSE; no significant differences among groups were observed in ER-alpha uterine stromal levels. After treatment with 17beta-estradiol, estrogen receptor-beta (ER-beta) expression was not modulated in the stroma or epithelium, whereas the SSE induced an up-regulation of ER-beta stromal expression. Collectively, these results suggest that the lack of stimulatory activity on the uterine epithelium using soy treatment could be due to a negligible stimulatory activity on estrogen receptor-alpha and/or to the enhanced expression observed in stromal ER-beta, the latter being considered as a negative modulator of ERalpha-mediated uterine proliferation. 17beta-estradiol, but not the SSE, down-regulated uterine epithelial progesterone receptor (PR), compared with ovariectomized rats. In the stromal compartment, progesterone receptor expression was fully up-regulated by 17beta-estradiol treatment and, to a lesser extent, by SSE treatment. The minor increase in lipid levels induced by ovariectomy was not affected by SSE administration. Finally, the lack of stimulatory activity on uterus was also confirmed in an immature female rat model. CONCLUSIONS: The results of this study demonstrated that the tested extract has an interesting profile of tissue-specific response, in that it is efficacious in preventing experimental osteoporosis without causing stimulation in uterus at doses that are effective in bone.     

Soy isoflavones for osteoporosis: an evidence-based approach

Effects of soy isoflavones on osteoporosis remain unclear. This review aimed to clarify the effect of soy isoflavones on bone mineral density (BMD) and turnover markers in menopausal women. PubMed and the Cochrane Library were searched in July 2011 for relevant meta-analyses of randomized controlled trials evaluating effects of soy isoflavones on BMD and bone turnover markers. Three meta-analyses evaluated the effects of soy isoflavones on lumbar spine, total hip, femoral neck, and trochanter BMD. Soy isoflavones significantly improved lumbar spine BMD in a moderate manner, but did not affect total hip, femoral neck, and trochanter BMD in menopausal women. Ingestion of soy isoflavones for six months appeared to be enough to exert a beneficial effect on lumbar spine BMD. Two meta-analyses evaluated the effects of soy isoflavones on a bone resorption marker (urine deoxypyridinoline) and two formation markers (serum alkaline phosphatase and osteocalcin). Soy isoflavones significantly decreased urine deoxypyridinoline in a moderate manner, but did not affect serum alkaline phosphatase and osteocalcin in menopausal women. Soy isoflavones may prevent postmenopausal osteoporosis and improve bone strength thus decreasing risk of fracture in menopausal women by increasing lumbar spine BMD and decreasing bone resorption marker urine deoxypyridinoline. Further studies are needed to address factors affecting the magnitude of the beneficial effects of soy isoflavones and to assess the possible interactions between soy isoflavones and anti-osteoporosis drugs, and to verify effects on BMD of other skeletal sites and other bone turnover markers.     

Marked Individual Variation in Isoflavone Metabolism After a Soy Challenge Can Modulate the Skeletal Effect of Isoflavones in Premenopausal Women

Soy-isoflavones may act as estrogenic agonists or antagonists depending on the endogenous hormone status. These clinical effects can be exerted variably in individuals by the metabolic ability to produce a more potent metabolite than precursors. The objective of this randomized, double-blind, placebo-controlled study was to investigate the skeletal effect of isoflavones according to their metabolic variability in premenopausal women. Volunteers were randomly assigned to receive either soy-extract isoflavones (n=32) or lactose (n=21) once a day for three menstrual cycles. After intervention, the urinary excretions of isoflavones and their metabolites were significantly higher in the soy group than in the placebo group and showed a large inter-individual variation. Women in the soy group were divided into subgroups according to their ability to excrete more potent metabolites. Serum osteocalcin and urine deoxypyridinoline showed a tendency to increase after a challenge in equol high-excretors. Serum osteocalcin concentration in the genistein high-excretors increased significantly after a challenge (P=0.04) but did not increase in either the placebo or genistein low-excretors. An estrogenic antagonistic effect of isoflavones on bone turnover was observed in premenopausal women who are able to produce more potent metabolites.     

The effect of soy protein isolate on bone metabolism

OBJECTIVE: This double-blind, 15-month pilot study was designed to investigate the effect of soy protein isolate with varying concentrations of isoflavones on early postmenopausal bone loss and lipids. DESIGN: A total of 65 women, with a mean age of 55 years and 7.5 years since menopause, were randomized to one of three groups; soy protein with 96 mg isoflavones/day, soy with 52 mg isoflavones/day, or soy without isoflavones (< 4 mg isoflavones/day). Soy was given for 9 months and then discontinued; participants were followed for an additional 6 months. Bone mineral density (BMD) and blood lipids were measured during this time. RESULTS: Measurement of serum isoflavones at 3 months showed dose-related increases in the three groups. There was no significant effect of the soy supplements on BMD of the spine or femoral neck in any of the three groups. BMD increased significantly in the trochanter at 9 months (P = 0.0219) and at 15 months (P < 0.05) in the group given isoflavone-free soy compared with the other two groups. There was no significant effect of soy on lipid metabolism at the end of the intervention. CONCLUSION: The present study did not find a significant positive effect of soy protein isolate supplemented with isoflavones on BMD and the serum lipid profile in early postmenopausal women.     

Effects of dietary isoflavone aglycones on the reproductive tract of male and female mice

We assessed the effects of dietary consumption of soy isoflavone aglycones on the reproductive tract of sexually mature male and female mice. Isoflavone concentrates with a ratio of 10:1, 2:1 or 1:10 genistein:daidzein (G:D) were added to provide 120 mg total isoflavones/1800 Calories (approximately 40 mg/kg body weight) to diets having either casein/lactalbumin or soy protein isolate as the source of protein. After 16 weeks, mice were necropsied and gross and histopathologic assessments of uterus, vagina, testes and accessory sex glands were completed. Effects of the 10G:1D isoflavone concentrates were absent or minimal in females but in males included atrophy of accessory sex glands. In contrast, the 2G:1D and 1G:10D concentrates caused dramatic estrogenic effects in both male and female mice. Effects in females included endometritis and effects typical of estrogenic stimulation (i.e., uterine enlargement, keratinization of vaginal epithelium, increased height of endometrial surface epithelial cells, and uterine squamous metaplasia). Effects in males included reduced plasma testosterone concentrations, atrophy of seminiferous epithelium, atrophy of accessory sex glands, and squamous metaplasia of seminal vesicles. Some effects varied with protein source. We conclude that a diet containing approximately 40 mg/kg soy isoflavone aglycones with a genistein:daidzein ratio of 2:1 or less has marked estrogenic effects on the reproductive system of male and female mice.     

The effects of fructo-oligosaccharides in combination with soy protein on bone in osteopenic ovariectomized rats

OBJECTIVE: The intestinal microflora is important in rendering soy isoflavones bioavailable by facilitating their conversion to equol. Hence, substances that can modulate the intestinal microflora could affect the bioavailability of isoflavones. In this study, we examined the effects of fructo-oligosaccharides (FOS), a prebiotic, on enhancing the effects of soy isoflavones on bone in ovariectomized osteopenic female rats. DESIGN: Sixty-three 9-month-old female Sprague-Dawley rats were either sham-operated (Sham; one group) or ovariectomized (Ovx; four groups) and were fed a control diet for 3 months to induce bone loss. After bone loss was confirmed via dual-energy x-ray absorptiometry, rats were placed on dietary treatment for 4 months. The Sham and one Ovx group received a control diet, and the remaining Ovx groups received either a soy protein-based diet (Soy), a FOS-supplemented diet (FOS), or a soy protein-based and FOS-supplemented diet (Soy+FOS). Before the termination of the study, whole-body bone mineral density (BMD) and bone mineral content (BMC) were assessed under anesthesia. Immediately after euthanasia, bone specimens were collected for the assessments of BMD, BMC, and biomechanical and microarchitectural properties. RESULTS: Whole-body BMD values were significantly higher in FOS and Soy+FOS groups compared with Ovx controls. The tibial BMC increased by 10%, 6%, and 4% in Soy, FOS, and Soy+FOS groups, respectively, compared to the Ovx control group. FOS and FOS+Soy treatments had the most pronounced effects in enhancing lumbar BMC and BMD. The FOS+Soy combination effectively improved tibial microarchitectural properties by enhancing trabecular number and lowering trabecular separation compared with Ovx controls. The effects of dietary treatments on lumbar microarchitectural properties were minimal and biomechanical properties of the femur were not affected by any of the dietary treatments. CONCLUSION: Our findings suggest that, although incorporation of either soy or FOS in the diet of Ovx rats can improve BMD of the whole body, tibiae, and lumbar vertebrae, their combination had no any additive effects. However, in terms of microarchitecture, the combination of soy and FOS had a greater effect in reversing the loss of certain microarchitectural parameters such as tibial trabecular number, separation, and thickness.     

去势大鼠骨形态计量学、生物性能变化

目的 通过观察大鼠血清学、骨形态计量学测定、骨生物性能等方法研究去卵巢大鼠骨变化.方法 采用7-12月龄SD雌性大鼠30只,随机分为对照组、模型组和尼尔雌醇组.模型组和尼尔雌醇组大鼠切除双侧卵巢进行造模.尼尔雌醇组大鼠造模后45天开始灌服尼尔雌醇,连续90天;对照组和模型组大鼠手术后第45天开始灌服去离子水,连续90天.所有大鼠给药第80、87天时两次ip四环素20mg/kg进行骨荧光标记.实验结束时,取出大鼠股骨及第四腰椎骨进行股骨形态计量学测定、股骨骨密度测量、股骨三点弯曲实验及椎骨压缩实验,同时测定血清碱性磷酸酶(ALP)、抗酒石酸酸性磷酸酶(StrACP)水平.结果 大鼠去双侧卵巢后,股骨骨密度、骨最大载荷、最大应力、弹性模量、刚度水平和骨小梁体积显著性提高;椎体压缩性能、股骨形态计量学、股骨弯曲性能有明显改变;而雌激素对以上变化有不同程度改善作用.结论 骨组织形态、骨生物性能是评价骨质疏松模型或药物疗效的重要指标;发生骨质疏松时,骨组织形态学变化先于骨生物性能变化.     

淫羊藿和女贞子提取物对骨质疏松症大鼠性激素功能水平的影响

目的:探讨淫羊藿和女贞子提取物对骨质疏松症(OP)大鼠性激素功能水平的影响。方法:50 只3 月龄雄性Wistar 大鼠随机分为正常组、模型组、提取物低剂量组、提取物中剂量组、提取物高剂量组共5 组,每组10 只,除正常组外,其他4 组以维甲酸灌喂15 d 制作OP 模型,造模成功后正常组和模型组灌服生理盐水,提取物低剂量组、中剂量组和高剂量组分别按50 mg/ kg、100 mg/ kg、200 mg/ kg 灌服淫羊藿和女贞子提取物混合物,均持续3 周。测量各组骨密度(BMD),左侧股骨切片采用苏木精和伊红(HE)染色观察股骨病理学改变并测定骨小梁组织形态相关参数,免疫组织化学分析激素受体蛋白表达。结果:正常组股骨全部、股骨上端、股骨下端BMD 显著高于模型组,提取物中剂量组和高剂量组股骨全部、股骨上端、股骨下端BMD 显著高于模型组,提取物低剂量组股骨全部BMD 显著高于模型组,差异均有统计学意义(P<0郾05 或P<0郾01);与模型组比较,提取物各组骨小梁数量有了显著增加,完整性、连续性明显好转,相关参数比较结果显示,正常组Tb.N、Tb.Ar、Tb.Th 显著大于模型组,Tb.Sp 显著小于模型组,提取物中剂量组和高剂量组Tb.N、Tb.Ar、Tb.Th 显著大于模型组,Tb.Sp 显著小于模型组,提取物低剂量组,Tb、Ar、Tb、Th 显著大于模型组,差异均具有统计学意义(P<0.05 或P<0.01);正常组AR、ER蛋白表达量明显高于模型组,而提取物高剂量组AR、ER 蛋白表达量较模型组显著上升,差异有统计学意义(P<0.05 或P<0.01)。结论:淫羊藿和女贞子提取物能够增加维甲酸诱导的OP 大鼠BMD,逆转骨组织病理结构,上调性激素受体蛋白表达,对维甲酸诱导的OP 大鼠具有潜在的保护作用。     

The effects of soy extract on the uterus of castrated adult rats

OBJECTIVE: The aim of this study was to evaluate the effects of different doses of a standardized soy extract on the uterus of castrated rats. METHODS: Fifty-six adult castrated female Wistar rats were randomly divided into seven groups (eight animals in each) that received: GI--drug vehicle (propylene glycol); GII--soy extract 10mg/kg per day; GIII--soy extract 50mg/kg per day; GIV--soy extract 100mg/kg per day; GV--soy extract 300mg/kg per day; GVI--soy extract 600mg/kg per day; GVII-conjugated equine estrogens (CEE) 200microg/kg per day. After 21 days of treatment, all animals were sacrificed and fragments of the uterine horns were immediately removed, fixed in 10% formaldehyde and submitted to routine histological techniques for morphometric study. The endometrial cell proliferation index was determined with the PCNA antibody PC-10 and expressed as the percentuals of the PCNA-positive nuclei relative to the total countings. Other fragments were immediately frozen in liquid nitrogen for RNA extraction and VEGF analysis using RT-PCR technique. RESULTS: The minimal dose of soy extract that produced a significant increase of the morphometric parameters was 100mg/kg (GIV). The maximum effects on endometrial and myometrial morphometry were detected in the groups treated with 300 and 600mg/kg of soy extract (groups V and VI) and CEE (GVII). The expression of PCNA in the endometrial epithelium and stroma was increased by treatment with 100-600mg/kg per day of soy extract (groups IV-VI) or with CCE (group VII). Doses equal to or higher than 50mg/kg of soy extract (groups III-VI) and CEE stimulated the expression of VEGF. CONCLUSION: The treatment of adult castrated rats during 21 days with doses of 100mg/kg per day or higher of soy extract may determine significant proliferation in the endometrium and myometrium.     

绝经后骨质疏松症模型体视学测量和机理初探

目的:观测去势术后大鼠骨组织体视学的改变,复制绝经后骨质疏松症的动物模型,初步探讨骨质疏松症的发病机制。方法:将31只3月龄雌性SD大鼠随机分为卵巢切除术组(OVX)和假手术组(sham),术后28d和56d分别处死。测量子宫湿重,骨矿物密度(BMD)和骨组织形态计量参数,分析骨组织微结构的变化。结果:OVX组术后28d和56d大鼠子宫湿重和股骨远端1/3处骨密度均显著少于sham组(P＜0.05);OVX组股骨远端和胫骨近端的干骺端骨小梁面积百分率显著少于sham组(P＜0.01)。结论:卵巢切除术后大鼠股骨远端骨矿物密度和骨组织形态计量学参数稳步下降,胫骨近端骨矿物密度和骨组织形态计量学参数迅速下降而且不稳定;雌激素减少导致的骨质疏松主要发生在长骨的干骺端,骨骺受影响较少;骨组织形态计量参数中骨小梁面积百分比敏感性和稳定性较高。     

Soy protein and isoflavone effects on vasomotor symptoms in peri- and postmenopausal women: the Soy Estrogen Alternative Study

OBJECTIVE: To investigate the efficacy of dietary soy proteins containing differing amounts of isoflavones on the number and severity of vasomotor symptoms (hot flashes and night sweats) in peri- and postmenopausal women. DESIGN: A double-masked, randomized, controlled, clinical trial was conducted. A total of 241 community-dwelling women reporting vasomotor symptoms at baseline were randomized into one of three groups. In all groups, participants consumed a daily supplement containing 25 g of soy protein and were randomly assigned to one of three groups: (a) isoflavone extracted soy protein (control), (b) soy protein with a medium dose of isoflavones (42 mg/day), or (c) soy protein with a higher dose of isoflavones (58 mg/day). The primary outcome measure in this trial was change in reported vasomotor symptoms. RESULTS: A reduction in the number and severity of vasomotor symptoms was observed in all three treatment groups. No significant differences in the number and severity of vasomotor symptoms were observed among the high isoflavone, middle isoflavone, or control groups. The lack of a between-treatment group effect was observed even after stratified by number of baseline symptoms and use of traditional hormone replacement therapy. CONCLUSIONS: These data suggest that soy protein containing 42 or 58 mg of isoflavones is no more effective than isoflavone-extracted soy protein for improving the number and severity of vasomotor symptoms in peri- and postmenopausal women.     

Effect of soy protein-containing isoflavones on lipoproteins in postmenopausal women

OBJECTIVE: Some clinical trials have demonstrated a beneficial effect of dietary soy protein on improving lipoproteins. Research also has documented that serum lipoproteins and some lipoprotein subclasses are altered as a consequence of menopause, resulting in a more atherogenic lipid profile. The purpose of this study was to determine the effects of isolated soy protein-containing isoflavones on lipoproteins and lipoprotein subclasses in both African American and white postmenopausal women with borderline to moderate low-density lipoprotein cholesterol elevations. DESIGN: This was a randomized, double-blind, controlled clinical trial including 216 postmenopausal women. After a 4-week run-in period with a casein protein-based supplement, participants were randomly assigned to continue the casein placebo or receive soy protein-containing isoflavones for a period of 12 weeks. RESULTS: In the soy group, the total cholesterol, low-density lipoprotein cholesterol, and low-density lipoprotein particle number decreased significantly as compared with the placebo group at 6 weeks. Although this decrease continued at 12 weeks in the soy group, the difference from the placebo group was attenuated for total cholesterol and low-density lipoprotein particle number. Multivariate analyses controlling for age, race, change in weight, change in dietary fat intake, and change in kilocalorie energy expenditure revealed that treatment remained a significant independent predictor of change in total cholesterol (P = 0.01), low-density lipoprotein cholesterol (P = 0.02), and low-density lipoprotein particle number (P = 0.002) after 6 weeks of dietary soy. CONCLUSIONS: Increased consumption of soy protein replacing animal protein that is high in fat may help improve atherogenic lipid profiles.     

A comparison of oral micronized estradiol with soy phytoestrogen effects on tail skin temperatures of ovariectomized rats

OBJECTIVE: Whether phytoestrogen-containing soy supplements have beneficial effects on hot flashes of postmenopausal women and how those effects, if any, compare to estrogen replacement therapy has been uncertain. It is possible that the uncertainty is due to the low doses of soy isoflavones (30-60 mg per day) used in the studies. We used ovariectomized retired breeder rats and a higher dose of soy phytoestrogens to approach these uncertainties experimentally. DESIGN: The treatment groups were as follows: (1) Control group fed a casein/lactalbumin-based diet; (2) Soy(-) group fed alcohol-washed soy protein isolate with the phytoestrogens extracted; (3) Soy(+) group fed phytoestrogen-containing soy protein (equivalent to a woman's dose of 144 mg isoflavones per day)--a dose two to three times higher than that in most studies with women; and (4) E2 group fed oral micronized estradiol (E2) at a dose equivalent to a woman's dose of 1 mg per day. A temperature-transponder was taped to the surface of the tail to measure temperature. Tail skin temperature was significantly increased within a week after ovariectomy. The animals were pair-fed during the last 21 days of treatment for daily temperature measurement. RESULTS: Soy(-) had no effect on skin temperature. E2 had a large effect on skin temperature (about 1.4 degrees C reduction from Control). Soy(+) was intermediate between the E2 treatment and no treatment (about 0.8 degrees C reduction from Control). CONCLUSIONS: Soy phytoestrogens have a modest effect on average skin temperatures, being about half that of E2, even at high doses in the rat model.     

Soy isoflavones modulate the expression of BAD and neuron-specific beta III tubulin in male rat brain

Isoflavones, the most abundant phytoestrogens in soy foods, are structurally similar to 17beta-estradiol. There is evidence that soy isoflavones influence neuronal apoptosis or proliferation in vitro and in vivo. However, little research has been done to investigate the effects of soy isoflavones on markers of neuronal apoptosis and survival in vivo. We examined brain BAD (a proapoptotic member of Bcl-2 protein family) and neuron-specific beta III tubulin (an early marker of neuronal differentiation/survival) levels in male rats fed either a standard chow rich of soy isoflavones (Phyto-600) or one free of soy isoflavones (Phyto-free) life-long (from conception until time tissue collected). Among five brain regions, the expression of BAD was highest in medial basal hypothalamus (MBH); the next highest in hippocampus; moderate in amygdala and frontal cortex; and lowest in cerebellum in Phyto-free fed animals. In animals on Phyto-600 diet, the levels of BAD were significantly decreased in frontal cortex and MBH; but significantly increased in the amygdala. The expression of beta III tubulin was highest in frontal cortex; moderate in amygdala, hippocampus and MBH; and lowest in cerebellum in the Phyto-free group. In rats fed with the Phyto-600 diet, levels of beta III tubulin were significantly increased in amygdala, frontal cortex, hippocampus and MBH compared to Phyto-free values. In summary, these findings provide evidence for the neuroprotective potential of soy isoflavones in the amygdala, frontal cortex, hippocampus and MBH. This implies that consumption of soy isoflavones may be beneficial on learning and memory, anxiety-related behaviors, and recovery from trauma.     

Protective effect of plant extract from Onobrychis ebenoides on ovariectomy-induced bone loss in rats

OBJECTIVE: Certain plant extracts have been the object of recent studies due to their mild estrogenic action and their possible potential role in osteoporosis prevention and/or treatment. The present study was undertaken to investigate the possible protective effect of the aqueous solution of the plant Onobrychis ebenoides, with proven in vitro mild estrogenic action, on bone mass loss of the ovariectomized (Ovx) rat experimental model of osteoporosis. METHODS: Forty intact female mature (10-month-old) Wistar rats were separated into three groups: Ovx, Ovx plus plant extract (Ph) and sham-operated (control). Ph administration in the drinking water at a dose of 300 mg/kg body weight/day commenced immediately after Ovx. Bone mineral density (BMD) values, percentage change from the baseline measurement and histomorphometry of the tibia, as well as body and uterine weight, were examined and compared between groups. RESULTS: Comparison of BMD absolute values of the whole tibia of Ovx + Ph and Ovx animals at both 3 and 6 months post-Ovx were highly significant (p < 0.0005), showing a protective effect on treated animals. The extract did not appear to have such a beneficial effect on BMD of the proximal tibia of the treated animals compared to the Ovx animals after 3 months; however, a significant protective effect was observed at 6 months post-Ovx in treated animals compared to the Ovx (p = 0.015). When the % changes from baseline measurement of the whole tibia of Ovx + Ph and controls were compared, there was no significant difference at 3 or 6 months, demonstrating a highly protective effect; the respective comparisons of proximal tibia % changes did not display such protection. Body and uterine weight comparisons showed no significant difference between Ovx and treated rats, whereas, the level of significance for each group compared to controls was p < 0.0005. CONCLUSIONS: The Ph studied showed a highly significant protective effect on BMD of the whole tibia of Ovx rats after 3 and 6 months of treatment, compared to the non-treated animals. Its effect on the proximal tibia was less pronounced, but also statistically significant compared to non-treated rats after 6 months. The lack of significant effect on body and uterine weight is in favor of its selective estrogen receptor modulator-like activity, and merits further studies.     

Preferential vascular-based transfer from vagina to the corpus but not to the tubal part of the uterus in postmenopausal women.

BACKGROUND: Vaginal administration of progesterone during infertility treatment has therapeutic advantages over oral administration. However, the reasons for this are poorly defined. To demonstrate a preferential vagina-to-uterus distribution of substances, we investigated cold distribution from vagina to the uterus and rectum. METHOD: In 10 postmenopausal women, thermoprobes were inserted into the uterine cavity and in the rectum at <9 cm or at >9 cm from the anus; temperatures were subsequently measured during 10 min flushing of vagina with cold saline. RESULTS: After 10 min, temperature decreased as follows: uterus, tubal angle: -0.22 +/- 0.07 degrees C, 10 (mean +/- SEM, n); uterus, middle cavity: -1.26 +/- 0.34 degrees C, 9; rectum, <9 cm insertion: -3.69 +/- 0.68 degrees C, 3; rectum, >9 cm insertion: -0.51 +/- 0.19 degrees C, 6. CONCLUSIONS: Despite obviously different distances to the vagina of the uterine and the low rectal probes (<9 cm) the temperature decrease occurred at the same time. Cold transfer from vagina to the uterus and rectum is probably not the result of simple diffusion but of a vascular counter-current transfer. Differential cooling of corpus and tubal angles suggests a different arterial supply; while uterine corpus is supplied from the uterine artery, the tubal angles seem to be mainly supplied from the ovarian artery via the tubal arcade.