固本增骨方对去卵巢骨质疏松模型大鼠血清BALP及NEI网络组织中CaM、CaMKⅡ mRNA表达的影响

目的观察固本增骨方对去卵巢骨质疏松模型大鼠下丘脑、垂体、脾脏、肾上腺和股骨组织中钙调蛋白（CaM） mRNA、CaM依赖性蛋白激酶Ⅱ（CaMK） mRNA表达以及血清骨碱性磷酸酶（BALP）的影响，探讨固本增骨方通过神经-内分泌-免疫（NEI）网络调节防治骨质疏松的机制。方法采用双侧去卵巢法建立大鼠骨质疏松模型，随机分为模型对照组、戊酸雌二醇组、固本增骨方高、中、低浓度组，分别予以生理盐水、戊酸雌二醇、不同剂量（9.2、4.6、2.3 g/kg）生药灌胃；选取同龄雌性大鼠作为空白对照组（仅灌胃生理盐水，不造模）。干预后，分别在第4、8、12、16周时各组随机抽取10只大鼠，心脏取血，ELISA法检测血清BALP含量，分别于动物模型成功后，灌胃干预4、8、12、16周后采用实时荧光定量-聚合酶链式反应（RT-PCR）检测下丘脑、垂体、肾上腺、脾脏、股骨组织中CaM、CaMKⅡ mRNA表达。结果在干预的不同时相点，模型组大鼠血清BALP及NEI网络下丘脑、垂体、肾上腺、脾脏、股骨组织CaM、CaMKⅡ mRNA表达与空白对照组差异有统计学意义（ P＜0.05）。与模型对照组相比，戊酸雌二醇组和固本增骨方组血清BALP质量浓度及下丘脑、垂体组织CaM mRNA表达升高（ P＜0.05），而肾上腺、脾脏、股骨组织CaM mRNA表达及下丘脑、垂体、肾上腺、脾脏、股骨组织CaMKⅡ mRNA表达降低，差异有统计学意义（ P<0.05）；与戊酸雌二醇组相比较除固本增骨方低剂量组股骨和肾上腺组织中对CaM、CaMKⅡ mRNA表达的调节作用稍差（ P<0.05）外，戊酸雌二醇与固本增骨方各组在其余各组织对CaM、CaMK Ⅱ具有相同程度的调节作用。结论固本增骨方可提高去卵巢骨质疏松模型大鼠的血清BALP质量浓度，调节去卵巢骨质疏松模型大鼠NEI网络中CaM、CaMKⅡ mRNA的表达，从而防治骨质疏松。

英文摘要:

Objective To observe the effect of Guben Zenggu Decoction on the expressions of calmodulin (CaM) and calcium/calmodulin-dependent protein kinase Ⅱ (CaMK Ⅱ) mRNA in the hypothalamus, pituitary, spleen, adrenal gland and femur in ovariectomized rats. To explore the mechanism of Guben Zenggu Decoction regulating and controlling osteoporosis through neuro-endocrine-immune (NEI) network. Methods The osteoporosis model was established by ovariectomy and randomly divided into model control group, Estradiol valerate group, Guben Zenggu Decoction high, medium and low concentration group. Selected the same age rats as the blank control group, after intervention, 10 rats were randomly selected at the time of 4th, 8th, 12th and 16th weeks from each group, After the success of the model and the successful intervention of Guben Zenggu Decoction for 4, 8, 12 and 16 weeks, the blood were taken from the heart, the serum bone alkaline phosphatase (BALP) levels was measured by the enzyme-linked immunosorbent assay (ELISA), and the thalamus, pituitary, spleen, adrenal, femoral tissue of rats were collected to detected the expressions of CaM and CaMKⅡ mRNA by using real-time fluorescence quantitative polymerase chain reaction (RT-PCR). Results At different time points (4 weeks, 8 CM（155mm]weeks, 12 weeks, 16 weeks), the expressions of serum BALP and CaM and CaMKⅡ mRNA in NEI network in model group were significantly different from those in blank control group ( P<0.05). Compared with the model control group, serum BALP concentration and CaM mRNA expression in hypothalamus and pituitary tissue were significantly increased in estradiol valerate and Guben Zenggu Decoction groups ( P<0.05). However, the expression of CaM mRNA in adrenal gland, spleen and femur and the expression of CaMKⅡ mRNA in hypothalamus, pituitary, adrenal gland, spleen and femur were significantly decreased ( P<0.05); compared with estradiol valerate group, the effect of Guben Zenggu Decoction on the expressions of CaM mRNA and CaMKⅡ mRNA in femoral and adrenal tissues was significantly different ( P<0.05). Conclusion Guben Zenggu Decoction can increase the serum concentration of BALP, regulate the expression of CaM mRNA and CaMKⅡ mRNA in hypothalamus, pituitary, adrenal gland, spleen and femur, thus play a role in prevention and treatment of osteoporosis by regulating NEI network.

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