肾阳虚COPD合并骨质疏松大鼠肠道菌群、OPG/RANKL/RANK通路变化及益生菌的治疗作用

Changes of intestinal flora,OPG/RANKL/RANK pathway in kidney-yang deficiency COPD combined with osteoporosis and therapeutic effect of probiotics in rats

目的研究肾阳虚慢性阻塞性肺疾病(COPD)合并骨质疏松大鼠肠道菌群、骨保护素(OPG)/核因子κB受体活化因子配体(RANKL)/核因子κB受体活化因子(RANK)通路变化及益生菌的治疗作用。方法雄性SD大鼠(8~10周龄、体重180~200 g)随机分为对照组、模型组、益生菌组,每组8只。模型组和益生菌组大鼠进行肾阳虚COPD合并骨质疏松建模,建模成功后模型组给予生理盐水灌胃、益生菌组给予益生菌[0.9×10^(9)CFU/(g•kg•d)]灌胃,持续28 d。检测肺功能用力肺活量(FVC)、第0.2秒用力呼气量(Fev 0.2),股骨的骨密度(BMD),血清中干扰素-γ(IFN-γ)、白介素-4(IL-4)、碱性磷酸酶(ALP)、Ⅰ型前胶原氨基端原肽(PINP)、抗酒石酸酸性磷酸酶(TRAP)、Ⅰ型胶原羧基端肽β特殊序列(β-CTX)的水平,肺组织和骨组织中OPG、RANK、RANKL的mRNA表达水平。结果模型组的FVC、Fev 0.2、BMD、血清ALP和PINP水平以及肠道菌群丰富度的Ace指数、Chao1指数、Shannon指数、肺组织和骨组织中OPG的mRNA表达水平均低于对照组,血清IFN-γ/IL-4比例、IFN-γ水平、TRAP水平、β-CTX水平、肺组织和骨组织中RANK与RANKL的mRNA表达水平均高于对照组(均P<0.05);益生菌组的FVC、Fev 0.2、BMD、血清ALP和PINP水平以及肠道菌群丰富度的Ace指数、Chao1指数、Shannon指数、肺组织和骨组织中OPG的mRNA表达水平均高于模型组,血清IFN-γ/IL-4比例、IFN-γ水平、TRAP水平、β-CTX水平、肺组织和骨组织中RANK与RANKL的mRNA表达水平均低于模型组(均P<0.05)。结论肾阳虚COPD合并骨质疏松大鼠存在肠道菌群失调和OPG/RANKL/RANK通路异常,补充益生菌显著纠正肠道菌群失调、OPG/RANKL/RANK通路异常并改善肺功能和骨密度。

Objective To observe the changes of intestinal flora and osteoprotegerin(OPG)/receptor activator for nuclear factor-κB ligand(RANKL)/receptor activator for nuclear factor-κB(RANK)pathway,and the therapeutic effect of probiotics in rats with kidney-yang deficiency chronic obstructive pulmonary disease(COPD)complicated with osteoporosis.Methods Male SD rats were randomly divided into control group,model group and probiotic group,with 8 rats in each group.Kidney-yang deficiency COPD complicated with osteoporosis rat models were established in model group and probiotic group.The model group was given normal saline while the probiotic group was given a probiotic[0.9×10^(9) CFU/(g•kg•d)]for 28 days.Pulmonary function forced vital capacity(FVC),forced expiratory volume 0.2 second(Fev 0.2),bone mineral density(BMD)of femur,serum levels of interferon-γ(IFN-γ),interleukin-4(IL-4),alkaline phosphatase(ALP),procollagenⅠN-terminal propeptide(PINP),tartrate resistant acid phosphatase(TRAP)andβC-terminal cross-linked telopeptides of type I collagen(β-CTX),the mRNA expression levels of OPG,RANK and RANKL in lung tissue and bone tissue were measured.Results The levels of FVC,Fev 0.2,BMD,serum ALP and PINP contents,Ace index,Chaol index,Shannon index of intestinal flora richness,OPG mRNA expression in lung tissue and bone tissue in model group were lower,while the proportion of serum IFN-γ/IL-4,contents of IFN-γ,TRAP,β-CTX,mRNA expression levels of RANK and RANKL in lung and bone tissues were higher than those in control group,respectively(all P<0.05).The levels of FVC,Fev 0.2,BMD,serum ALP and PINP contents,Ace index,Chaol index,Shannon index of intestinal flora richness,OPG mRNA expression in lung tissue and bone tissue in probiotic group were higher,while the proportion of serum IFN-γ/IL-4,contents of IFN-γ,TRAP,β-CTX,mRNA expression levels of RANK and RANKL in lung and bone tissues were lower than those in the model group,respectively(all P<0.05).Conclusion Rats with Kidney-yang deficiency COPD combined with osteoporosis have intestinal flora imbalance and abnormal OPG/RANKL/RANK pathway.Supplementation of probiotics can significantly correct intestinal flora imbalance and abnormal OPG/RANKL/RANK pathway,and improve lung function and bone mineral density.