摘要
目的 观察马钱子苷(Loganin)干预去卵巢(OVX)小鼠骨量、肠道菌群及神经肽的变化.方法 28只雌性C57BL/6J小鼠随机分为假手术组(SHAM)、OVX组、Loganin低剂量组(Loganin-L)和Loganin高剂量组(Loganin-H),每组7只.Loganin-L组和Loganin-H组在OVX基础上分别予Loganin 5 mg/(kg·d)和20 mg/(kg·d)灌胃,SHAM组和OVX组灌胃等量蒸馏水,每日1次,持续12周.实验结束后处死取材,Micro-CT检测左侧股骨BMD、BV/TV、Tb.N及Tb.Sp;ELISA法检测血清中CTX、TRAP、P1NP、TNF-α、IL-6、IL-10、LPS、CGRP、SP、VIP、NPY水平;16 S rRNA高通量测序技术检测粪便中肠道菌群多样性与差异性.结果 OVX 组 BMD、BV/TV、Tb.N、P1NP、IL-10、CGRP、SP、VIP、NPY 较 SHAM 组显著降低(P＜0.05,P＜0.01),Tb.Sp、CTX、TRAP、TNF-α、IL-6、LPS 显著增高(P＜0.05,P＜0.01).Loganin-L 组 BMD、BV/TV、Tb.N、P1NP、IL-10、CGRP、SP、VIP、NPY 较 OVX 组有升高趋势(P＞0.05),Tb.Sp、CTX、TRAP、TNF-α、IL-6、LPS 有降低趋势(P＞0.05).Loganin-H 组 BMD、BV/TV、Tb.N、P1NP、IL-10、CGRP、SP、VIP、NPY 较 OVX 组显著增高(P＜0.05),Tb.Sp、CTX、TRAP、TNF-α、IL-6、LPS 显著降低(P＜0.05).在门水平,与 SHAM 组相比,OVX 组 Firmicutes/Bacteroidota(F/B)率显著降低(P＜0.05),Loganin-L 组 F/B 率较OVX组有升高趋势(P＞0.05),Loganin-H组F/B率显著升高(P＜0.05);在属水平,OVX组norank_f_Muribaculaceae丰度较SHAM 组显著增加(P＜0.05),Lactobacillus 丰度显著减少(P＜0.05);较 OVX 组,Loganin-L 组 norank_f_Muribaculaceae 丰度有降低趋势(P＞0.05),Lactobacillus 丰度有升高趋势(P＞0.05),Loganin-H 组 norank_f_Muribaculaceae 丰度显著减少(P＜0.05),Lactobacillus丰度显著增加(P＜0.01).结论 Loganin呈剂量依赖性抑制OVX小鼠骨量丢失,其机制可能是通过调节肠道菌群中norank_f_Muribaculaceae和Lactobacillus丰度、增加神经肽含量及降低炎症反应实现的.
Abstract
Objective To observe the changes of bone mass,gut microbiota and neuropeptides in ovariectomized mice treated with Loganin.Methods 28 female C57BL/6J mice were randomly divided into sham operation group(SHAM),ovariectomized group(OVX),Loganin low-dose group(Loganin-L)and Loganin high-dose group(Loganin-H),with 7 mice in each group.Loganin-L and Loganin-H groups received Loganin 5 mg/(kg·d)and 20 mg/(kg·d)via oral gavage,respectively,while SHAM and OVX groups received the same dose of distilled water via oral gavage.After 12 weeks of intervention,BMD,BV/TV,Tb.N and Tb.Sp of the left femur were detected by Micro-CT.CTX,TRAP,P1NP,TNF-α,IL-6,IL-10,CGRP,SP,VIP,NPY and LPS were detected by ELISA.16 S rRNA high-throughput sequencing was used to detect the gut microbiota.Results BMD,BV/TV,Tb.N,P1NP,IL-10,CGRP,SP,VIP and NPY were significantly decreased in OVX group compared with SHAM group(P＜0.05,P＜0.01);Tb.Sp,CTX,TRAP,TNF-α,IL-6 and LPS were significantly increased in OVX group(P＜0.05,P＜0.01).BMD,BV/TV,Tb.N,P1NP,IL-10,CGRP,SP,VIP,and NPY of Loganin-L group tended to increase compared with OVX group(P＞0.05),while Tb.Sp,CTX,TRAP,TNF-α,IL-6 and LPS tended to decrease(P＞0.05).BMD,BV/TV,Tb.N,P1NP,IL-10,CGRP,SP,VIP and NPY were significantly increased in Loganin-H group compared with OVX group(P＜0.05);Tb.Sp,CTX,TRAP,TNF-α,IL-6 and LPS were significantly decreased(P＜0.05).At the phylum level,F/B ratio was significantly decreased in OVX group(P＜0.05)compared with SHAM group.However the F/B ratio in Loganin-L group has an increasing trend compared with OVX group(P＞0.05),and it was significantly increased in Loganin-H group(P＜0.05).At the genus level,norank_f_Muribaculaceae exhibited a significant increase(P＜0.05)while Lactobacillus demonstrated a significant decrease(P＜0.05)in OVX group compared with SHAM group.Compared with OVX group,norank_f_Muribaculaceae tended to decrease in Loganin-L group(P＞0.05),and Lactobacillustended to increase(P＞0.05).Norank_f_Muribaculaceae showed a significant decrease(P＜0.05)whereas Lactobacillus exhibited a significant increase(P＜0.01)in Loganin-H group.Conclusion Loganin can inhibit bone loss in OVX mice in a dose-dependent manner,which may be achieved by reducing inflammatory response,increasing the level of neuropeptides and regulating the abundance of norank_f_Muribaculaceae and Lactobacillus.
维普
万方数据