摘要
目的 基于炎性因子和肠道菌群的变化,探讨牛蒡子苷(ARC)抑制钛颗粒诱导小鼠颅骨周围骨溶解的机制.方法 20 只 7 周龄C57BL/6J雌性小鼠被随机均匀分配到以下组:空白组(SHAM)、骨溶解模型组(Vehicle)、ARC低剂量组(L-ARC)以及ARC高剂量组(H-ARC),每组 5 只.L-ARC、H-ARC组在Vehicle组基础上分别给予 5、10 mg/(kg·d)ARC腹腔注射,SHAM组和Vehicle组腹腔注射等剂量生理盐水,每日 1 次,共 14 d.通过Micro-CT测定小鼠颅骨BMD、BV/TV、Tb.N、BS/BV、Tb.Sp和Tb.Th;ELISA法检测小鼠血清中TRACP5b、CTX、OCN、IL-6、TNF-α和IL-10 水平;16S rRNA高通量测序技术分析粪便中菌群结构与丰度变化.结果 Vehicle组BMD、BV/TV、Tb.N、BS/BV、Tb.Th、OCN和IL-10 较SHAM组显著降低(P＜0.01,P＜0.05),Tb.Sp、TRACP5b、CTX、IL-6和TNF-α显著升高(P＜0.05,P＜0.01);L-ARC组BMD、BV/TV、Tb.N、BS/BV、Tb.Th和OCN较Vehicle组有升高趋势(P＞0.05),Tb.Sp、TRACP5b和CTX有降低趋势(P＞0.05),TNF-α和IL-6 显著降低(P＜0.05),IL-10 显著升高(P＜0.05);H-ARC组BMD、BV/TV、Tb.N和IL-10 较Vehicle组显著升高(P＜0.05,P＜0.01),TRACP5b、CTX、IL-6和TNF-α显著降低(P＜0.05,P＜0.01),BS/BV、Tb.Th和OCN有升高趋势(P＞0.05),Tb.Sp有降低趋势(P＞0.05).在门水平,Vehicle组Firmicutes/Bacteroidota(F/B)率较SHAM组有升高趋势(P＞0.05);L-ARC和H-ARC组F/B率较Vehicle组显著降低(P＜0.01).在属水平,Vehicle组Ileibacterium丰度较SHAM组有升高趋势(P＞0.05),Prevotellaceae_UCG-001 和Rikenellaceae_RC9_gut_group丰度有降低趋势(P＞0.05);较Vehicle组,经ARC干预后,Ileibacterium丰度显著降低(P＜0.05),Prevotellaceae_UCG-001 和Rikenellaceae_RC9_gut_group丰度显著增加(P＜0.05).结论 ARC可通过抑制炎性反应、调节肠道菌群结构抑制钛颗粒诱导的小鼠颅骨骨溶解,且呈剂量依赖性.
Abstract
Objective To investigate the mechanism of inhibiting titanium partical-induced osteolysis around the skull with arctiin(ARC)in mice based on the changes of inflammatory factors and gut microbiota.Methods Twenty 7-week-old C57BL/6J female mice were randomly and evenly assigned to the following groups:blank group(SHAM),model group(Vehicle),ARC low-dose group,(L-ARC)and ARC high-dose group(H-ARC).During the following 14 days,mice in the experimental group were injected intraperitoneally with two different concentrations of ARC[5 and 10 mg/(kg·d)],respectively.Mice in the sham group and vehicle group were injected with 0.9%NaCl every day.The following parameters of skulls,BMD,BV/TV,Tb.N,BS/BV,Tb.Sp,and Tb.Th were detected with micro-CT.The levels of TRACP5b,CTX,OCN,IL-6,TNF-α,and IL-10 in serum were detected with ELISA.16 S rRNA sequencing was used to analyze the structure and abundance of gut microbiota.Results BMD,BV/TV,Tb.N,BS/BV,Tb.Th,OCN,and IL-10 decreased significantly in Vehicle group compared to those in SHAM group(P＜0.01,P＜0.05).Tb.Sp,TRACP5b,CTX,IL-6,and TNF-α significantly increased(P＜0.05,P＜0.01).Compared to those in Vehicle group,BMD,BV/TV,Tb.N,BS/BV,Tb.Th,and OCN in L-ARC group had an increasing trend(P＞0.05),Tb.Sp,TRACP5b,and CTX had a decreasing trend(P＜0.05),TNF-α and IL-6 significantly decreased(P＜0.05),and IL-10 significantly increased(P＜0.05).Compared to those in Vehicle group,the levels of BMD,BV/TV,Tb.N,and IL-10 in H-ARC group significantly increased(P＜0.05,P＜0.01),the levels of TRACP5b,CTX,IL-6,and TNF-α significantly decreased(P＜0.05,P＜0.01),while the levels of BS/BV,Tb.Th,and OCN had an increasing trend(P＞0.05),and Tb.Sp had a decreasing trend(P＞0.05).At the phylum level,compared to that in SHAM group,firmicutes/bacteroidota(F/B)ratio had an increasing trend in Vehicle group(P＞0.05).However,compared to that in Vehicle group,the F/B ratio in L-ARC and H-ARC groups significantly decreased(P＜0.01).At the genus level,compared to that in SHAM group,the abundance of ileibacterium in Vehicle group had an increasing trend(P＞0.05),the abundance of prevotellaceae_UCG-001 and rikenellaceae_RC9 in gut in Vehicle group had a decreasing trend(P＞0.05).While compared to that in Vehicle group,the abundance of ileibacterium in ARC groups significantly decreased(P＜0.05),the abundance of prevotellaceae UCG-001 and rikenellaceae RC9 in gut in ARC groups significantly increased(P＜0.05).Conclusion ARC inhibits titanium partical-induced osteolysis in mice by suppressing inflammation and modulating the structure of gut microbiota in a dose-dependent manner.
维普
万方数据