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基于16S核糖体RNA测序研究健脾方对绝经后骨质疏松症小鼠肠道菌群的影响 被引量:1

A study of the effectsof Jianpi Fang(健脾方)on intestinal flora of mice with postmenopausal osteoporosis based on 16S ribosomal RNA gene sequencing
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摘要 目的:探讨健脾方对绝经后骨质疏松症(osteoporosis,OP)小鼠肠道菌群的影响。方法:将30只雌性C57BL/6小鼠随机分为假手术组、模型组和健脾方组,每组10只。模型组和健脾方组小鼠通过双侧卵巢摘除术进行OP造模,假手术组小鼠打开腹腔后切除卵巢附近的一团脂肪组织,保留卵巢。造模后第7天,假手术组和模型组小鼠以生理盐水灌胃,健脾方组以健脾方药液灌胃(中药配方颗粒用量为54mg·kg^(-1)),3组小鼠均每天灌胃1次,每次0.2mL,连续灌胃28d。药物干预结束后,测定小鼠体质量,获取各组小鼠粪便样本、左侧股骨,切取子宫并称重,进行小鼠肠道菌群16S核糖体RNA测序分析及小鼠股骨病理学观察。结果:①一般情况。实验期间各组均无小鼠死亡。药物干预前,3组小鼠体质量的差异无统计学意义[(20.88±0.83)g,(20.65±0.74)g,(20.59±0.47)g,F=0.484,P=0.622]。药物干预结束后,模型组小鼠的体质量高于假手术组和健脾方组[(26.96±1.63)g,(23.42±0.84)g,P=0.000;(26.96±1.63)g,(24.62±1.55)g,P=0.001],假手术组和健脾方组小鼠体质量的差异无统计学意义(P=0.063)。假手术组小鼠的子宫质量高于模型组和健脾方组[(0.66±0.05)g,(0.24±0.05)g,P=0.000;(0.66±0.05)g,(0.28±0.06)g,P=0.000],模型组和健脾方组小鼠子宫质量的差异无统计学意义(P=0.053)。②小鼠肠道菌群丰度分析结果。假手术组与模型组含有499个共同操作分类单元(operational taxonomic units,OTUs)数据;除共同OTUs数据外,假手术组有87个特异性OTUs数据,模型组有74个特异性OTUs数据。模型组与健脾方组含有489个共同OTUs数据;除共同OTUs数据外,模型组有84个特异性OTUs数据,健脾方组有54个特异性OTUs数据。偏最小二乘法判别分析结果显示,3组小鼠肠道菌群丰度差异明显。③小鼠肠道菌群结构分析结果。物种累计曲线随样本量增加逐渐趋于平缓,说明样品中的物种已经基本被测序覆盖,测 Objective:To explore the effects of Jianpi Fang(健脾方,JPF)on intestinal flora of postmenopausal osteoporosis(PMOP)mice.Methods:Thirty female C57BL/6 mice were randomly divided into shamoperated group,model group and JPF group,10 mice in each group.The mice in model group and JPF group were subjected to bilateral ovariectomy for inducing PMOP;while the ones in shamoperated group underwent removal of periovarian adipose tissues,with ovaries retained.On the 7th day after the modeling operation,the mice in shamoperated group and model group were intragastric administrated with normal saline,while the ones in JPF group with JPF decoction(the dosage of Chinese medicinal granules was 54 mg/kg).All mice in the 3 groups were intragastric administrated once a day,0.2 mL at a time for consecutive 28 days.After the end of drug intervention,the body mass of the mice was measured and their stool samples and left femurs were harvested,moreover their uteri were dissected and weighed,and analysis on 16S ribosomal RNA(rRNA)gene sequencing of intestinal flora and pathological observation on femurs of the mice were conducted.Results:①No dead mice were found during the experiment period.There was no statistical difference in body mass among the 3 groups before the drug intervention(20.88±0.83,20.65±0.74,20.59±0.47 g,F=0.484,P=0.622).After the end of drug intervention,the body mass was higher in model group compared with that of shamoperated group and JPF group(26.96±1.63 vs 23.42±0.84 g,P=0.000;26.96±1.63 vs 24.62±1.55 g,P=0.001),while the difference in body mass between shamoperated group and JPF group was not statistically significant(P=0.063).The quality of uteri was higher in shamoperated group compared with that of model group and JPF group(0.66±0.05 vs 0.24±0.05 g,P=0.000;0.66±0.05 vs 0.28±0.06 g,P=0.000),while there was no significant difference between model group and JPF group(P=0.053).②The shamoperated group and model group shared 499 common operational taxonomic units(OTUs).In addition to the common OTUs
作者 陈斯亮 石瑛 汤辰明 马威 詹红生 CHEN Siliang;SHI Ying;TANG Chenming;MA Wei;ZHAN Hongsheng(Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)
出处 《中医正骨》 2023年第6期11-17,23,共8页 The Journal of Traditional Chinese Orthopedics and Traumatology
基金 国家自然科学基金项目(81674003) “华东片区及市级中医专科筋骨病损专科联盟建设”项目。
关键词 骨质疏松 绝经后 胃肠道微生物群 RNA 核糖体 16S 健脾方 osteoporosis,postmenopausal gastrointestinal microbiome RNA,Ribosomal,16S Jianpi fang
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