摘要
目的观察强骨抗萎膏对尾吊大鼠骨细胞凋亡和内质网应激的影响,探讨强骨抗萎膏防治模拟失重状态下骨丢失的机制。方法60只SD大鼠分为对照组、模拟失重组、强骨抗萎膏组、阿仑膦酸钠组,骨疏康颗粒组,每组12只。采用尾部悬吊模拟失重状态,尾吊28 d;对照组和模拟失重组给予每日等体积生理盐水灌胃,其它3组复制模型,分别每日给予中药强骨抗萎膏2.4 g/kg,阿仑膦酸钠0.007 g/kg,骨疏康颗粒0.26 g/kg灌胃。持续给药4周后处死大鼠,TUNEL法检测骨细胞凋亡情况;Western blot法检测葡萄糖调节蛋白78(GRP78)和线粒体融合蛋白2(MFN2)表达,RT-PCR和Western blot检测骨组织中蛋白激酶R样内质网激酶(PERK),肌醇依赖性激酶1α(IRE1α)、活化转录因子6(ATF6)及C/EBP同源蛋白(CHOP)的基因和蛋白表达。结果TUNEL结果显示模拟失重组骨细胞凋亡较对照组增加,强骨抗萎膏可抑制骨细胞凋亡。与对照组大鼠比较,模拟失重组PERK基因表达量明显增高(P<0.05),强骨抗萎膏组较模拟失重组PERK基因表达量下降(P<0.05);与对照组比较,模拟失重组的GRP78、PERK、CHOP和ATF6蛋白含量明显增高(P<0.05),MFN2蛋白表达降低(P<0.05),强骨抗萎膏组较模拟失重组GRP78、PERK、CHOP和ATF6蛋白含量均明显降低(P<0.05),MFN2表达量升高。结论强骨抗萎膏可以改善模拟失重大鼠骨细胞凋亡,通过抑制骨组织内质网应激相关因子GRP78、PERK、ATF6和CHOP,上调MFN2表达,从而改善失重条件下的骨丢失。
Objective To observe the effects of the Qiang Gu Kang Wei Extraction on bone apoptosis and endoplasmic reticulum stress in tail suspension rats,and to explore the mechanism of the extraction in preventing bone loss in simulated weightlessness.Methods 60 SD rats were randomly divided into control group,simulated weightlessness group,Qiang Gu Kang Wei Extraction group,alendronate sodium group and Gushukang granule group,with 12 rats in each group.The tail suspension was used to simulate weightlessness for 28 days.The control group and the simulated weightlessness group were given equal volume of normal saline,while the other three groups were tail suspended and given 2.4 g/kg Qiang Gu Kang Wei Extraction,0.007 g/kg alendronate sodium,and 0.26 g/kg Gushukang granule by gavage,respectively.After 4 weeks of intragastric administration,rats were sacrificed,and the apoptosis of bone cells was detected by TUNEL.RT-PCR detected transcription levels of protein kinase R-like endoplasmic reticulum kinase(PERK),inositol requiring enzyme 1 alpha(IRE1α),activating transcription factor 6(ATF6),C/EBP homologous protein(CHOP)in bone tissue.Western blot was used to detect the protein expression of glucose-regulated protein 78(GRP78),PERK,IRE1α,ATF6,CHOP and mitofusin 2(MFN2).Results TUNEL results showed that Qiang Gu Kang Wei Extraction inhibited apoptosis.RT-PCR results showed that,compared with the control group,the content of PERK were significantly higher in the simulated weightlessness rgoup(P<0.05),while the PERK in the Qiang Gu Kang Wei Extraction group were significantly lower(P<0.05).Western blot results showed that,compared with the control group,the content of GRP78,PERK,CHOP and ATF6 were significantly higher and MFN2 was decreased in the simulated weightlessness rgoup(P<0.05),while the GRP78,PERK,CHOP and ATF6 were significantly lower and MFN2 was increased in the Qiang Gu Kang Wei Extraction group(P<0.05).Conclusion The Qiang Gu Kang Wei Extraction can reduce bone cells apoptosis of simulated weightlessness rats through inhibiting GRP78,PERK,ATF6 and CHOP,as well as upregulating MFN2 expression in bone tissue,thereby reducing bone loss under weightlessness condition.
作者
史长华
田雷瑜
冯婧
李姝玉
刘军莲
刘雨梦
刘嘉鹏
王谦
SHI Changhua;TIAN Leiyu;FENG Jing;LI Shuyu;LIU Junlian;LIU Yumeng;LIU Jiapeng;WANG Qian(Beijing University of Chinese Medicine,Beijing 100029,China;China Astronaut Research and Training Center,Beijing 100094,China)
出处
《云南中医药大学学报》
2023年第4期93-99,共7页
Journal of Yunnan University of Chinese Medicine
基金
载人航天工程航天医学实验领域项目(HYZHXM05002)。
关键词
失重
骨丢失
内质网应激
线粒体融合蛋白2
强骨抗萎膏
weightlessness
bone loss
endoplasmic reticulum stress
MFN2
Qiang Gu Kang Wei Extraction
