摘要
目的 探讨免疫调节分子IL-33对绝经后骨质疏松症(postmenopausal osteoporosis,PMOP)模型鼠RANKL/c-Fos/NFATc1信号轴的调控作用及藤黄健骨胶囊(Tenghuang Jiangu capsule,TJC)的干预机制.方法 构建PMOP大鼠模型,设置假手术组、PMOP模型组、阳性对照组(0.09 mg/kg)和TJC高、中、低剂量组(0.36、0.18、0.09 g/kg),灌胃给药,每天1次,持续8周.从大鼠末次给药后体质量、骨密度(bone mineral density,BMD)及股骨组织微结构等方面评价TJC的疗效;通过ELISA分析TJC对大鼠血清免疫调节因子IL-33、IL-1、IL-31变化;运用qPCR和Western blotting分析TJC对大鼠股骨OPG、RANKL、RANK、c-Fos、NFATc1表达量的影响.结果 与假手术组相比,PMOP模型组大鼠的体质量、骨髓脂肪组织相对面积(relative area of bone marrow adipose tissue,BMAT),IL-1、IL-31 变化,RANKL、RANK、c-Fos、NFATc1 的表达均出现了明显升高的趋势,而BMD、IL-33变化、OPG的表达则出现了明显下降的趋势(P＜0.01);与PMOP模型组相比,TJC高剂量组大鼠体质量出现显著下降趋势、IL-33变化出现显著升高趋势(P＜0.01),TJC中、高剂量组大鼠BMAT、RANK的表达出现显著下降趋势、OPG的表达出现显著升高趋势(P＜0.05或P＜0.01),TJC各剂量组大鼠IL-1、IL-31变化,RANKL、c-Fos、NFATc1出现显著降低趋势,BMD出现显著增加趋势(P＜0.05或P＜0.01).结论 TJC能够提高PMOP大鼠免疫调控分子IL-33含量,抑制免疫调控分子IL-1、IL-31含量和破骨细胞分化标志分子NFATc1的表达,其机制可能与RANKL/c-Fos/NFATc1抑制骨代谢通路密切相关.
Abstract
Objective To explore regulation effects of immunomodulatory molecule IL-33 on RANKL/c-Fos/NFATc1 signal axis in rats with postmenopausal osteoporosis(PMOP)and intervention mechanism of Tenghuang Jiangu capsule(TJC).Methods The rat's PMOP model was performed by bilateralovariectomy method.The rats were divided into sham group,PMOP model group,positive control group(0.09 mg/kg),TJC high,middle and low dose group(0.36,0.18,0.09 g/kg).These groups were intragastric administration once a day for 8 weeks.The curative effect of TJC was evaluated including body mass after last administration,BMD,and bone micro architecture.And the levevls of immune regulatory factors IL-33,IL-1,and IL-31in rat serum was checked by ELISA,the expressions of OPG,RANKL,RANK,c-Fos,NFATc1 in rat thigh-bone was determined by qPCR and Western blotting.Results Compared with the sham group,the body mass,BMAT,the levels of IL-1,IL-31,the expression of TRANKL,RANK,c-Fos,NFATc1 in the PMOP model group have a significant increase trend,the content of BMD,the level of IL-33 and the expression of OPG appeared a significant decrease trend(P＜0.01).Compared with the PMOP model group,the body mass appeared significantly decrease trend in the TJC high dose group,and the level of IL-33 has a increase trend(P＜0.01).BMAT,the expression of RANK appeared significantly decrease trend in the TJC middle dose group and the TJC high dose group,the expression of OPG appeared significantly increase trend(P＜0.05 or P＜0.01).The levels of IL-1,IL-31,the expression of TRANKL,RANK,c-Fos,NFATc1 have a significant decrease trend in the TJC low dose group,the TJC middle dose group and the TJC high dose group,the BMD has a significant increase trend(P＜0.05 or P＜0.01).Conclusion TJC can significantly raise the concentration of IL-33 in PMOP model rats,decrease the concentration ofIL-1,IL-31,and inhibit the expression of osteoclast differentiation marker NFATc1.Its mechanism may be closely related to the inhibition of RANKL/c-Fos/NFATc1 bone metabolic pathway.
基金项目
国家自然科学基金(82060872)
甘肃省中医药局项目(GZKP-2023-39)
甘肃省中医药局项目(GZKP-2023-63)
兰州市卫生健康科技发展项目(2021004)
兰州市科技计划(2022-3-22)
甘肃省"双一流"科研重点项目(GSSYLXM-05)
维普
万方数据