基于肠道菌群与肠道代谢探究脉络舒通丸对大鼠股骨骨折引起的后肢肿胀的治疗作用及机制  

Therapeutic effect and mechanism of Mailuo Shutong Pills on posterior limb swelling caused by femur fracture in rats based on intestinal flora and intestinal metabolism

作　　者：杨澜 刘明飞 孙成宏 项海鑫 苗宇 程国良 YANG Lan;LIU Ming-fei;SUN Chenghong;XIANG Hai-xin;MIAO Yu;CHENG Guo-liang(School of Pharmacy,Shandong University of Traditional Chinese Medicine,Ji'nan 250355,China;Key Laboratory of Marine Materia Medica,School of Medicine,Ocean University of China,Qingdao 266100,China;State Key Laboratory of Generic Technology of Chinese Traditional Medicine Pharmaceutical,Lunan Pharmaceutical Group Co.,Ltd.,Linyi 276005,China)

机构地区：[1]山东中医药大学药学院,山东济南250355 [2]中国海洋大学医学院海洋药物重点实验室,山东青岛266100 [3]鲁南制药集团股份有限公司中药制药共性技术国家重点实验室,山东临沂276005

出　　处：《中国中药杂志》2023年第17期4711-4721,共11页China Journal of Chinese Materia Medica

基　　金：山东省自然科学基金项目(ZR2022LZY021)。

摘　　要：探讨脉络舒通丸对大鼠股骨骨折引起的后肢肿胀的保护作用及其潜在机制。将大鼠随机分为假手术组、模型组、脉络舒通丸(MLST)低剂量组(1.8 g·kg^(-1)·d^(-1))、MLST高剂量组(3.6 g·kg^(-1)·d^(-1))和阳性药组(迈之灵片60 mg·kg^(-1)·d^(-1))。假手术组暴露股骨后缝合伤口,其余4组均进行机械性损伤造成股骨骨折,于造模前7 d及造模后5 d对各给药组给予灌胃给药,假手术组、模型组给予等剂量蒸馏水灌胃。采用苏木精-伊红(hematoxylin-eosin,HE)染色检测大鼠后肢肌肉组织的病理损伤,并测量其后肢肿胀度。酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)试剂盒检测各组大鼠血清中白细胞介素-6(interleukin-6,IL-6)、白细胞介素^(-1)β(interleukin^(-1)β,IL^(-1)β)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的水平,以及超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)、过氧化氢酶(catalase,CAT)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活性。蛋白免疫印记(Western blot)技术检测大鼠后肢肌肉组织中血红素氧合酶1(heme oxygenase 1,HO-1)、醌氧化还原酶NADH 1[NAD(P)H quinone oxidoreductase 1,NQO1]、核转录因子E2相关因子2(nuclear transcription factor E2-related factor 2,Nrf2)蛋白表达水平。通过16S rDNA测序和超高效液相色谱-串联质谱(UPLC-MS/MS)检测大鼠肠道菌群及肠道代谢物的变化进而挖掘出MLST治疗大鼠股骨骨折引起的后肢肿胀的潜在机制。与模型组相比,MLST可明显改善大鼠后肢肿胀程度;降低血清炎性因子水平;减轻氧化应激损伤。HE染色结果表明,MLST组大鼠后肢肌肉组织中炎性浸润情况显著改善。Western blot结果显示,与模型组相比,MLST可显著升高大鼠后肢肌肉组织中HO-1、NQO1、Nrf2蛋白表达。16S rDNA测序结果表明,MLST可以改善股骨骨折后大鼠肠道菌群的紊乱;UPLC-MS/MS结果表明,MLST显著影响股骨骨折This study aimed to investigate the protective effect and underlying mechanism of Mailuo Shutong Pills(MLST)on posterior limb swelling caused by femur fracture in rats.The rats were randomly divided into a sham operation group,a model group,a low-dose MLST group(1.8 g·kg^(-1)·d^(-1)),a high-dose MLST group(3.6 g·kg^(-1)·d^(-1)),and a positive drug group(60 mg·kg^(-1)·d^(-1)Maizhiling Tablets).The femur in the sham operation group was exposed and the wound was sutured,while the other four groups underwent mechanical damage to cause femur fracture.The rats were treated with corresponding drugs by gavage 7 days before modeling and 5 days after modeling,while those in the sham operation group and the model group were given an equivalent dose of distilled water by gavage.Hematoxylin-eosin(HE)staining was used to detect the pathological injury of the posterior limb muscle tissues in rats,and the degree of hind limb swelling was measured.The enzyme-linked immunosorbent assay(ELISA)kit was used to detect the expression levels of interleukin-6(IL-6),interleukin-1β(IL-1β),and tumor necrosis factor-α(TNF-α)in the serum of rats in each group.The activity of superoxide dismutase(SOD),malondialdehyde(MDA),catalase(CAT),and glutathione peroxidase(GSH-Px)in rat serum was also measured.Western blot was used to detect the protein expression levels of heme oxygenase 1(HO-1),NAD(P)H quinone oxidoreductase 1(NQO1),and nuclear transcription factor E2-related factor 2(Nrf2)in rat posterior limb muscle tissues.The changes in the intestinal flora and intestinal metabolites in rats were detected by 16S rDNA sequencing and ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS),respectively,to explore the underlying mechanism of MLST in treating posterior limb swelling caused by femur fracture in rats.Compared with the model group,MLST significantly improved the degree of posterior limb swelling in rats,reduced the levels of serum inflammatory factors,and alleviated oxidative stress injury.The HE staining res

关 键 词：脉络舒通丸 骨折肿胀 肠道菌群 非靶向代谢组学 

分 类 号：R285.5[医药卫生—中药学]