壮骨止痛胶囊调控Notch通路对去卵巢大鼠Notch1、Jagged1、HES蛋白的影响

Effect of Zhuanggu Zhitong Capsule(壮骨止痛胶囊)on Notch1,Jagged1 and HES Protein in Ovariectomized Rats

目的观察壮骨止痛胶囊对去势雌鼠Notch1、Jagged1、HES蛋白的表达差异,探究其治疗骨质疏松的具体机制。方法将3月龄的健康雌性SD大鼠60只,随机分为空白组(KBZ)、假手术组(JSSZ)、模型组(MXZ)、壮骨止痛胶囊组(ZGZTFZ)、雷帕霉素(RAPA)组、自噬抑制剂(3-MA)组,每组各10只。除空白组、假手术组外,其余各组建立绝经后骨质疏松模型,假手术组去除卵巢周围相应体积脂肪。灌胃1个月后取材制备各组血清。ELISA测定血清中Ⅰ型胶原氨基端延长肽(P1NP)含量;HE染色、免疫组化测定骨组织和肾组织中Notch1、Jagged1、HES的表达。结果与假手术组比较,模型组蛋白表达有明显差异(P<0.01);与模型组比较,壮骨止痛胶囊组和RAPA组Notch1、Jagged1、HES表达升高(P<0.01);与RAPA组比较,壮骨止痛胶囊组骨组织HES表达降低(P<0.01),Notch1表达升高(P<0.01),Jagged1表达差异无显著统计学意义(P>0.05),肾组织中HES、Notc-h1、Jagged1表达显著降低(P<0.01);3-MA组HES、Notch1、Jagged1表达显著降低(P<0.01)。结论壮骨止痛胶囊能调控去卵巢大鼠的Notch通路对Notch1、Jagged1、HES蛋白表达起正向调节作用,促进骨形成,干预绝经后骨质疏松的状况。

Objective To observe the expression differences of Zhuanggu Zhitong Capsule(壮骨止痛胶囊)on Notch1,Jag⁃ged1 and HES protein in castrated female mice and explore the specific mechanism of Zhuanggu Zhitong Capsule in the treatment of osteoporosis.Methods Sixty healthy female SD rats aged 3 months were randomly divided into blank group,sham operation group,model group,bone-building analgesic prescription group,RAPA group and autophagy inhibitor group(3-MA),with 10 rats in each group.In addition to the blank group and the sham operation group,the postmenopausal osteoporosis model was established in the other groups,and the corresponding volume fat around the ovary was removed in the sham operation group.After 1 month of gavage,the serum of each group was prepared.ELISA detected the serum collagen typeⅠamino terminal extension of the peptide(P1NP)content.The expressions of Notch1,Jagged1 and HES in bone and kidney were determined by HE staining and immunohistochemistry.Results The expressions of Notch1,Jagged1 and HES were increased in the Zhuanggu Zhitong Capsule group and RAPA group(P<0.01).Compared with those of the RAPA group,the expression of HES and the expression of Notch1 in Zhuanggu Zhitong Capsule group were decreased(P<0.01),and the expression of Jagged1 was not significantly different(P>0.05).The expressions of HES,Notch1 and Jagged1 in 3-MA group were significantly decreased(P<0.01).Conclusion Zhuanggu Zhitong Capsule can regulate Notch pathway in ovariectomized rats and positively regulate the expressions of Notch1,Jagged1 and HES proteins,promote bone formation and interfere with postmenopausal osteoporosis.