急性肺损伤模型中连花清瘟胶囊对巨噬细胞趋化能力的药效与机制研究
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摘要
该文探讨了连花清瘟胶囊(LHQW)在细菌感染所致的急性肺损伤(ALI)模型中,对病理性炎症的保护性作用以及潜在的作用机制。在体外实验中,分别检测RAW264.7细胞和THP-1细胞单核巨噬细胞内MCP-1 mRNA的表达和细胞上清中MCP-1的含量以及LHQW对巨噬细胞趋化能力的影响;在体内实验中,将小鼠随机分为正常组,模型(LPS 5 mg·kg~(-1))组,LHQW低(300 mg·kg~(-1))、中(600 mg·kg~(-1))、高(1 200 mg·kg~(-1))剂量组,地塞米松5 mg·kg~(-1)剂量组,青链霉素组7组,分别检测2 h小鼠肛温,小鼠肺组织干重和湿重,检测肺泡灌洗液中的TNF-α和MCP-1和血清中的MCP-1;观察肺泡巨噬细胞的浸润情况;并且采用CCK-8法检测了肺泡巨噬细胞的浸润数量;HE染色观察小鼠肺组织炎症浸润情况。体外实验和体内实验都证实了LHQW能够通过降低MCP-1表达量和单核巨噬细胞在肺部感染灶的趋化和募集,进而降低了病理性炎症损伤水平,阻断了ALI模型动物的疾病进展。
This paper was mainly to discuss the potential role and mechanism of Lianhua Qingwen Capsules(LHQW) in inhibiting pathological inflammation in the model of acute lung injury caused by bacterial infection. For in vitro study, the mRNA expression of MCP-1 in RAW264.7 cells and THP-1 cells, the content of MCP-1 in cell supernatant, as well as the effect of LHQW on chemotaxis of macrophages were detected. For in vivo study, mice were randomly divided into 7 groups, including normal group, model group(LPS 5 mg·kg~(-1)), LHQW 300, 600 and 1 200 mg·kg~(-1)(low, middle and high dose) groups, dexamethasone 5 mg·kg~(-1) group and penicillin-streptomycin group. Then, the anal temperature was detected two hours later. Dry weight and wet weight of lung tissues in mice were determined; TNF-α and MCP-1 levels in alveolar lavage fluid and MCP-1 in serum were detected. In addition, the infiltration of alveolar macrophages was also observed and the infiltration count of alveolar macrophages was measured by CCK-8 method. HE staining was also used to observe the inflammatory infiltration of lung tissues in mice. Both of the in vitro and in vivo data consistently have confirmed that: by down-regulating the expression of MCP-1, LHWQ could efficiently decrease the chemotaxis of monocytes toward the pulmonary infection foci, thus blocking the disease development in ALI animal model.
This paper was mainly to discuss the potential role and mechanism of Lianhua Qingwen Capsules(LHQW) in inhibiting pathological inflammation in the model of acute lung injury caused by bacterial infection. For in vitro study, the mRNA expression of MCP-1 in RAW264.7 cells and THP-1 cells, the content of MCP-1 in cell supernatant, as well as the effect of LHQW on chemotaxis of macrophages were detected. For in vivo study, mice were randomly divided into 7 groups, including normal group, model group(LPS 5 mg·kg~(-1)), LHQW 300, 600 and 1 200 mg·kg~(-1)(low, middle and high dose) groups, dexamethasone 5 mg·kg~(-1) group and penicillin-streptomycin group. Then, the anal temperature was detected two hours later. Dry weight and wet weight of lung tissues in mice were determined; TNF-α and MCP-1 levels in alveolar lavage fluid and MCP-1 in serum were detected. In addition, the infiltration of alveolar macrophages was also observed and the infiltration count of alveolar macrophages was measured by CCK-8 method. HE staining was also used to observe the inflammatory infiltration of lung tissues in mice. Both of the in vitro and in vivo data consistently have confirmed that: by down-regulating the expression of MCP-1, LHWQ could efficiently decrease the chemotaxis of monocytes toward the pulmonary infection foci, thus blocking the disease development in ALI animal model.
引文
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[2] Hazem E Ghoneim,Paul G Thomas,Jonathan A Mc Cullers.Depletion of alveolar macrophages during influenza infection facilitates bacterial super-infections[J].J Immunol,2013,191(3):1250.
[3] Boyton R J,Reynolds C J,Quigley K J,et al.Immune mechanisms and the impact of the disrupted lung microbiome in chronic bacterial lung infection and bronchiectasis[J].Clin Exp Immunol,2012,171:117.
[4] 崔雯雯,金鑫,张彦芬,等.连花清瘟胶囊对脂多糖致急性肺损伤小鼠IKK/IκB/NF-κB信号通路的影响[J].中成药,2015,37(5):953.
[5] 何园,张硌,刘慧莹,等.巨噬细胞对急性肺损伤后炎症的消解与组织修复的研究进展[J].中国呼吸与危重监护杂志,2018,17(4):430.
[6] McCullers J A.The co-pathogenesis of influenza viruses with bacteria in the lung[J].Nat Rev Microbiol,2014,12(4):252.
[7] Anuforom O,Wallace G R,Piddock L V.The immune response and antibacterial therapy[J].Med Microbiol Immunol,2015,204(2):151.
[8] Robinson K M,Kolls J K,Alcorn J F.The immunology of influenza virus-associated bacterial pneumonia[J].Curr Opin Immunol,2015,34:59.
[9] Rubenfeld G D,Caldwell E,Peabody E,et al.Incidence and outcomes of acute lung injury[J].N Engl J Med,2005,353(16):1685.
[10] 李波,唐思嘉,刘可可,等.姜黄素治疗急性肺损伤效果和机制的研究进展[J].吉林大学学报:医学版,2018,44(6):1312.