摘要
目的观察去卵巢骨质疏松症模型大鼠的骨组织和肾组织中Hedgehog信号通路细胞表面受体Patched-1(PTCH1)和Gli3mRNA和蛋白含量变化及补肾填精、活血化瘀中药复方对其影响,探讨中药组方治疗绝经后骨质疏松症的可行性以及绝经后骨质疏松症的发病机制与Hedgehog信号通路的关系。方法采用切除雌性大鼠双侧卵巢的方法建立绝经后骨质疏松症模型,分别给予补肾填精中药复方、活血化瘀中药复方、骨疏康对模型大鼠灌胃12周。然后将大鼠分为补肾填精组、活血化瘀组、阳性对照组、正常组和模型空白组,其中骨疏康作为阳性药物为阳性对照组。运用RT-PCR检测各组大鼠骨组织和肾组织PTCH1和Gli3mRNA相对表达量;通过ELISA法检测各组大鼠骨组织和肾组织PTCH1和Gli3蛋白含量。结果各组大鼠股骨和肾组织中PTCH1mRNA及蛋白表达方面:与正常组比较,模型组显著升高(P<0.01);与模型组比较,补肾填精组、活血化瘀组、阳性对照组显著降低(P<0.01);各个用药组间比较,补肾填精组下调优于活血化瘀组,具有显著性差异(P<0.01)。各组大鼠股骨和肾组织中Gli3mRNA及蛋白表达方面:与正常组比较,模型组显著升高(P<0.01);与模型组比较,补肾填精组、活血化瘀组、阳性对照组显著下降(P<0.01);各个用药组间比较,补肾填精组下调优于活血化瘀组,具有显著性差异(P<0.01)。结论PTCH1mRNA及蛋白含量和Gli3mRNA及蛋白含量在去卵巢骨质疏松症模型大鼠骨组织和肾组织中显著升高,提示PTCH1和Gli3可能参与绝经后骨质疏松症骨代谢失衡的发生和进展。补肾填精中药复方和活血化瘀中药复方可下调PTCH1和Gli3mRNA相对表达量及其蛋白含量,这可能是起到防治绝经后骨质疏松症的作用机制之一,并且补肾填精法优于活血化瘀法。
Objective To observe the changes of patched-1(PTCH1)and Gli3mRNA and protein contents of cell surface receptors of Hedgehog signaling pathway in bone and kidney tissues of ovariectomized osteoporosis model rats,as well as the effects of TCM prescriptions of tonifying kidney and filling essence,promoting blood circulation and removing blood stasis.To explore the feasibility of TCM prescription in the treatment of postmenopausal osteoporosis and the relationship between the pathogenesis of postmenopausal osteoporosis and Hedgehog signaling pathway.Me thods The model of postmenopausal osteoporosis was established by resecting the female rats’bilateral ovaries,and the model rats were given Chinese herbal formula of tonifying kidney and filling essence,Chinese herbal formula of promoting blood circulation and removing blood stasis,and Gushukang for 12 weeks,among which Gushukang was the positive control group,the normal group and the model blank group.The relative mRNA expression levels of PTCH1 and Gli3 in bone and kidney tissues of rats were detected by RT-PCR.The protein contents of PTCH1 and GLI3 in bone and kidney tissues of rats were determined by ELISA.Re sults Compared with the normal group,the expression of PTCH1 mRNA and protein in the femur and kidney of rats in each group was significantly increased in the model group(P<0.01).Compared with model group,it was significantly decreased in kidney-tonifying and essence filling group,blood circulation and stasis removing group and positive control group(P<0.01).Compared with each drug,the down-regulation of kidney-tonifying and essence filling group was better than the group of promoting blood circulation to remove blood stasis(P<0.01).Compared with the normal group,the expression of Gli3mRNA and protein in the femur and kidney of rats in each group was significantly increased in the model group(P<0.01).Compared with model group,it was significantly decreased in kidney-tonifying and essence filling group,blood circulation and stasis removing group and positive control group(P<0.01).Compared with each drug,the downregulation of kidney-tonifying and essence filling group was better than the group of promoting blood circulation to remove blood stasis(P<0.01).Conclusion The contents of PTCH1 mRNA and protein and GLI3 mRNA and protein were significantly increased in bone and kidney tissues of Ovariectomized rats with osteoporosis,suggesting that PTCH1 and GLI3 may be involved in the occurrence and progression of bone metabolic imbalance in postmenopausal osteoporosis.Kidney-tonifying and essence filling prescriptions and TCM prescriptions for promoting blood circulation and removing blood stasis can downregulate the mRNA relative expressions and protein contents of PTCH1 and GLI3,which may be one of the mechanisms of prevention and treatment of postmenopausal osteoporosis,and the Kidney-tonifying and essence filling method is better than the method for promoting blood circulation and removing blood stasis.
作者
邓洋洋
刘明欣
孙鑫
王庆谚
蒋宁
高巳东
王重一
林浩楠
郑洪新
DENG Yangyang;LIU Mingxin;SUN Xin;WANG Qingyan;JIANG Ning;GAO Sidong;WANG Zhongyi;LIN Haonan;ZHENG Hongxin(Liaoning University of TCM,Shenyang 110847,China)
出处
《中国骨质疏松杂志》
CAS
CSCD
北大核心
2021年第8期1112-1116,共5页
Chinese Journal of Osteoporosis
基金
国家重点研发计划(2018YFC1704301)
辽宁省自然科学基金项目(2020-MS-222)
辽宁省百千万人才工程(2017[59])
辽宁省教育厅“青年人才育苗”项目(L202024)。