左归丸对去势大鼠钙转运通路相关蛋白的影响

Effect of Zuoguiwan on Expression of Ca2+ Transport Pathway-related Protein in Ovariectomized Rats

目的:探讨左归丸对去势大鼠骨密度和骨组织中钙转运通路相关蛋白的影响。方法:选取48只雌性SD大鼠,随机分为正常组,假手术组,模型组,尼尔雌醇组(0.167 mg·kg^-1),左归丸低、高剂量组(9.6,38.4 g·kg^-1)。除假手术组和正常组,其余组切除大鼠卵巢,造模成功后3个月进行各组干预。3个月后处死大鼠,检测大鼠骨密度(BMD)和骨代谢标志物;蛋白免疫印迹法(Western blot)检测钙转运通路(骨组织)瞬时受体电位通道V5(TRPV5),钙结合蛋白-D28K(CaBP-D28K),钠钙交换器1(NCX1)和细胞膜钙汞(PMCA1b)蛋白表达。结果:与正常组比较,模型组血钙降低(P<0.01),与模型组比较,左归丸高剂量组和尼尔雌醇组大鼠血钙显著升高(P<0.01);与正常组比较,模型组BMD显著降低(P<0.01),与模型组比较,左归丸高剂量组与尼尔雌醇组大鼠BMD均有改善(P<0.05);与假手术组和正常组比较,模型组各蛋白表达均有不同程度上调(P<0.05,P<0.01),与模型组比较,左归丸高剂量组中TRPV5蛋白表达有显著下调(P<0.01),各干预组中的CaBP-D28K与NCX1蛋白表达均有不同程度的下调(P<0.05,P<0.01),PMCA1b蛋白表达在左归丸低剂量组和尼尔雌醇组内表达均有不同程度下调(P<0.05,P<0.01)。结论:左归丸可不同程度的降低大鼠破骨细胞中TRPV5,CAPB-D28K,PMCA1b,NCX1等介导破骨细胞吸收钙离子的重要通道蛋白的表达,左归丸的抗骨质疏松机理有可能是通过抑制TRPV5介导的破骨细胞的骨吸收而起作用。

Objective: To explore the effect of Zuoguiwan on the bone mineral density(BMD) and the expressions of Ca2 +transport-associated protein in ovariectomized rats.Method: The 48 female SD rats were randomly divided into six groups: normal group, model group, sham operation group, estrogen group(0.167 mg·kg^-1) and low and high-dose Zuoguiwan groups(9.6,38.4 g·kg^-1),with 10 rats in each group.Except for the sham-operated group,the ovariectomized rats in the other groups received the bilateral ovariectomy.Therapeutic intervention was given in each group for 3 months after the establishment of the model.After 12 weeks,BMD was measured using dualenergy X-ray absorptiometry.Tartrated presistant acid phosphatse(TRACP) and serum calcium were detected by biochemical kits.Protein expression in Ca^2+ transport(Bone tissue) was detected by Western blot.Result: Compared with the normal group,the serum calcium of the model group was decreased(P<0.01).Compared with the normal group,BMD of the model group was decreased(P<0.01).The serum calcium of rats in high-dose group and western medicine group was higher than that of model group(P<0.01).BMD in model group was lower than that of Zuoguiwan groups and estrogen group(P<0.05).There was no significant difference in TRACP among the groups.Nilestriol and Zuoguiwan can down-regulate the expressions of TRPV5,NCX1,CaBP-D28K Kand PMCA1 b in bone tissue of castrated rats(P<0.05,P<0.01).Conclusion:Zuoguiwan can down-regulate the expressions of Ca^2+ transport-associated proteins(Bone tissues) in rat osteoclasts,with an efficacy on osteoporosis.