更年安怡片对去卵巢雌鼠绝经后骨质疏松症防治作用的实验研究
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摘要
目的：
绝经后骨质疏松症(Postmenopausal Osteoporosis，PMO)是指绝经后妇女由于卵巢功能衰退，雌激素水平下降，从而在骨代谢过程中，骨吸收大于骨形成，出现以低骨量和骨组织的显微结构的退行性变为其特征，临床表现为骨脆性和骨折易感性增加的一种代谢性疾病。本课题在前期工作的基础上，运用生物免疫技术，从胰岛素样生长因子(IGF)调节角度，探讨更年安怡片预防和治疗绝经后骨质疏松症的作用机制，为临床预防和治疗绝经后骨质疏松症提供理论依据。
方法：
本实验选用雌性未孕SD大鼠60只为实验对象，5月龄，体重200-250g。将其中40只摘除双侧卵巢进行造模，10只行假手术，另10只不做处理作为正常对照组。造模的动物随机分为去卵巢对照组、雌激素组、更年安怡片预防组和更年安怡片治疗组，每组10只。更年安怡片预防组于术后7天开始用药，雌激素组和更年安怡片治疗组在术后90天开始用药，均治疗4个疗程。末次治疗结束后，麻醉下检测腰椎和股骨的骨矿含量(BMC)和骨密度(BMD)，摘眼球取血，离心取上清，采用放射免疫法和酶联免疫法分别检测雌二醇(E_2)和胰岛素样生长因子-Ⅰ(IGF-Ⅰ)。
结果：
1．去卵巢雌鼠腰椎及股骨的BMC和BMD均下降(P＜0.01)，血清E_2水平和IGF-Ⅰ含量也下降(P＜0.05)；
2．更年安怡片和雌激素均可提高去卵巢雌鼠腰椎和股骨的BMC、BMD，与去卵巢对照组比较有显著差异(P＜0.05)；
3．更年安怡片和雌激素均可以提高去卵巢大鼠血清E_2水平和IGF-Ⅰ含量，与去卵巢对照组比较有显著差异(P＜0.05)；
4．更年安怡片预防组和治疗组均可以提高腰椎和股骨的BMC和BMD，血清E_2水平和IGF-Ⅰ含量，但两者作用的效果不同(P＜0.05)，提示用药时机不同可能效果不同；
5．经相关性分析显示：血清IGF-Ⅰ含量和E_2水平呈正相关关系，相关系数r=0.9001，提示E_2通过IGF-Ⅰ是其发挥抗骨质疏松作用的另一途径。
结论：
更年安怡片通过上调血清E_2水平，提高IGF-Ⅰ含量，可以明显增高去卵巢大鼠腰椎和股骨的BMC和BMD，直接或间接作用于成骨细胞，增强骨重建，改善骨质疏松，为临床运用更年安怡片治疗绝经后骨质疏松症提供了理论依据。
Objective:
Postmenopausal Osteoporosis (PMO) is syndrome shown on women caused by the functional descent on both ovaries and the level of estrogen. During the metabolism of their bones, the bone absorptive rate is higher than formative rate. As the result, the progression of calcium deficiency and the degeneration of bone tissue are the characters in this period. Clinical manifestations appear on these elder women, which are osteosclerosis and increasing the possibility of fracture. The approach used here is based on the former fundamental research. By using the molecular biology technique, IGF system was tested to study the mechanism of the GNAT prophylaxis and treatment the PMO. By conducting experiment, effects of different treatments were compared to see whether the difference exists, and provide theoretical evident on clinical manifestation prophylaxis and treatment.
Methods:
In this experiment, 60 unfertilized female rats were chosen as the experimental objects, whose ages were all five-month-old and weight from 200 to 250 grams. By doing oophorectomy operations on 40 rats, fake operations on 10 rats, and non-operation on the rest of them as a normal group, the rats were randomly classified into four groups, 10 rats in each group. They were ovariectomized group, estrogen group, GNAT prophylaxis group, and GNAT treatment group. GNAT prophylaxis group started to take medicine 7 days after the operations. Both GNAT treatment and estrogen groups were conducted medicine treatments 90 days after the operations. All medicine treatments were four-course treatments. After the last treatment, the rats were anesthetized to test BMC (Bond Mineral Content) and BMD (Bond Mineral Density) and taken blood by excising the rats' eyeballs. The blood was centrifugated to get about 1ml serum to test E2 (Estradiol) and IGF- I (Insulin-like Growth Factor I ) respectively by RIA (Radioimmunoassay) and
ELISA (Enzyme-linked Immunoadsordent Assay).
Results:
1. Both BMC and BMD of ovariectomized rat's lumbar vertebra and thighbone decreased (P<0.01), so did the E2 and IGF-I (P<0.05);
2. Comparing with ovariectomized group, GANT and estrogen can increase BMC and BMD of the lumbar vertebra and thighbone (P<0.05);
3. Comparing with ovariectomized group, both GNAT and estrogen can enhance the level of serum E2 and IGF-I (P<0.05);
4. Both GNAT treatment group and GNAT prophylaxis group can increase BMC and BMD of the lumbar vertebra and thighbone, E2 and IGF-1 of the serum. Nevertheless, the results were different maybe due to the time of taking medicine which caused different effects.
5. According to relative analysis, the content of IGF-I and the E2 level of the
serum show obviously positive relationship, r = 0.9001 .It may indicated E2 played the role by IGF- I on osteoblast.
Conclusion:
According to the above results, both BMC and BMD of the ovariectomized rat's lumbar vertebra and thighbone decreased, at the same time, E2 and IGF-I decreased evidently and showed obviously positive relationship. GNAT not only can help the ovariectomized rats to increase BMC and BMD of the lumbar vertebra and thighbone, but also to increase the EI and IGF-I contents to form bone cells and improve osteoporosis problem. Compared with estrogen, GNAT lowers the negative side-effect, provides the entire comprehensive functions of traditional Chinese medicine, and provides new theory on clinical treatment.
绝经后骨质疏松症(Postmenopausal Osteoporosis，PMO)是指绝经后妇女由于卵巢功能衰退，雌激素水平下降，从而在骨代谢过程中，骨吸收大于骨形成，出现以低骨量和骨组织的显微结构的退行性变为其特征，临床表现为骨脆性和骨折易感性增加的一种代谢性疾病。本课题在前期工作的基础上，运用生物免疫技术，从胰岛素样生长因子(IGF)调节角度，探讨更年安怡片预防和治疗绝经后骨质疏松症的作用机制，为临床预防和治疗绝经后骨质疏松症提供理论依据。
方法：
本实验选用雌性未孕SD大鼠60只为实验对象，5月龄，体重200-250g。将其中40只摘除双侧卵巢进行造模，10只行假手术，另10只不做处理作为正常对照组。造模的动物随机分为去卵巢对照组、雌激素组、更年安怡片预防组和更年安怡片治疗组，每组10只。更年安怡片预防组于术后7天开始用药，雌激素组和更年安怡片治疗组在术后90天开始用药，均治疗4个疗程。末次治疗结束后，麻醉下检测腰椎和股骨的骨矿含量(BMC)和骨密度(BMD)，摘眼球取血，离心取上清，采用放射免疫法和酶联免疫法分别检测雌二醇(E_2)和胰岛素样生长因子-Ⅰ(IGF-Ⅰ)。
结果：
1．去卵巢雌鼠腰椎及股骨的BMC和BMD均下降(P＜0.01)，血清E_2水平和IGF-Ⅰ含量也下降(P＜0.05)；
2．更年安怡片和雌激素均可提高去卵巢雌鼠腰椎和股骨的BMC、BMD，与去卵巢对照组比较有显著差异(P＜0.05)；
3．更年安怡片和雌激素均可以提高去卵巢大鼠血清E_2水平和IGF-Ⅰ含量，与去卵巢对照组比较有显著差异(P＜0.05)；
4．更年安怡片预防组和治疗组均可以提高腰椎和股骨的BMC和BMD，血清E_2水平和IGF-Ⅰ含量，但两者作用的效果不同(P＜0.05)，提示用药时机不同可能效果不同；
5．经相关性分析显示：血清IGF-Ⅰ含量和E_2水平呈正相关关系，相关系数r=0.9001，提示E_2通过IGF-Ⅰ是其发挥抗骨质疏松作用的另一途径。
结论：
更年安怡片通过上调血清E_2水平，提高IGF-Ⅰ含量，可以明显增高去卵巢大鼠腰椎和股骨的BMC和BMD，直接或间接作用于成骨细胞，增强骨重建，改善骨质疏松，为临床运用更年安怡片治疗绝经后骨质疏松症提供了理论依据。
Objective:
Postmenopausal Osteoporosis (PMO) is syndrome shown on women caused by the functional descent on both ovaries and the level of estrogen. During the metabolism of their bones, the bone absorptive rate is higher than formative rate. As the result, the progression of calcium deficiency and the degeneration of bone tissue are the characters in this period. Clinical manifestations appear on these elder women, which are osteosclerosis and increasing the possibility of fracture. The approach used here is based on the former fundamental research. By using the molecular biology technique, IGF system was tested to study the mechanism of the GNAT prophylaxis and treatment the PMO. By conducting experiment, effects of different treatments were compared to see whether the difference exists, and provide theoretical evident on clinical manifestation prophylaxis and treatment.
Methods:
In this experiment, 60 unfertilized female rats were chosen as the experimental objects, whose ages were all five-month-old and weight from 200 to 250 grams. By doing oophorectomy operations on 40 rats, fake operations on 10 rats, and non-operation on the rest of them as a normal group, the rats were randomly classified into four groups, 10 rats in each group. They were ovariectomized group, estrogen group, GNAT prophylaxis group, and GNAT treatment group. GNAT prophylaxis group started to take medicine 7 days after the operations. Both GNAT treatment and estrogen groups were conducted medicine treatments 90 days after the operations. All medicine treatments were four-course treatments. After the last treatment, the rats were anesthetized to test BMC (Bond Mineral Content) and BMD (Bond Mineral Density) and taken blood by excising the rats' eyeballs. The blood was centrifugated to get about 1ml serum to test E2 (Estradiol) and IGF- I (Insulin-like Growth Factor I ) respectively by RIA (Radioimmunoassay) and
ELISA (Enzyme-linked Immunoadsordent Assay).
Results:
1. Both BMC and BMD of ovariectomized rat's lumbar vertebra and thighbone decreased (P<0.01), so did the E2 and IGF-I (P<0.05);
2. Comparing with ovariectomized group, GANT and estrogen can increase BMC and BMD of the lumbar vertebra and thighbone (P<0.05);
3. Comparing with ovariectomized group, both GNAT and estrogen can enhance the level of serum E2 and IGF-I (P<0.05);
4. Both GNAT treatment group and GNAT prophylaxis group can increase BMC and BMD of the lumbar vertebra and thighbone, E2 and IGF-1 of the serum. Nevertheless, the results were different maybe due to the time of taking medicine which caused different effects.
5. According to relative analysis, the content of IGF-I and the E2 level of the
serum show obviously positive relationship, r = 0.9001 .It may indicated E2 played the role by IGF- I on osteoblast.
Conclusion:
According to the above results, both BMC and BMD of the ovariectomized rat's lumbar vertebra and thighbone decreased, at the same time, E2 and IGF-I decreased evidently and showed obviously positive relationship. GNAT not only can help the ovariectomized rats to increase BMC and BMD of the lumbar vertebra and thighbone, but also to increase the EI and IGF-I contents to form bone cells and improve osteoporosis problem. Compared with estrogen, GNAT lowers the negative side-effect, provides the entire comprehensive functions of traditional Chinese medicine, and provides new theory on clinical treatment.
引文
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