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目的:研究喘可治注射液对脂多糖(LPS)诱导的小鼠急性肺损伤(ALI)的防治效果及作用机制。方法:SPF级雌性C57BL/6J小鼠48只,随机分为正常组、模型组、地塞米松组、喘可治组,每组12只。喘可治组每只小鼠腹腔注射0.2 m L·d-1,1次/d,连续7 d;地塞米松组造模前3 d按1 mg·kg~(-1)腹腔注射,1次/d,连续3 d;模型组注射等剂量的生理盐水,正常组不作处理。除正常组外,第8天腹腔注射LPS 0.1 m L(0.5 g·L~(-1)),乙醚吸入麻醉后,各小鼠滴鼻LPS 25μL(0.5 g·L~(-1)),制作ALI模型。24 h后取材检测。酶联免疫吸附法(ELISA)检测小鼠血清肿瘤坏死因子-α(TNF-α),白细胞介素-1β(IL~(-1)β),IL~(-1)8的含量;免疫组织化学染色检测核转录因子-κB(NF-κB)p65的蛋白表达;蛋白免疫印迹法(Western blot)检测肺组织p65,Toll样受体4(TLR4),Nod样受体蛋白3(NLRP3),半胱氨酸天冬氨酸蛋白酶-1(Caspase)-1蛋白表达;实时荧光定量PCR(Real-time PCR)检测肺组织p65和TLR4的mRNA表达。结果:与正常组比较,模型组小鼠血清TNF-α,IL~(-1)β,IL~(-1)8含量升高(P<0.01),肺组织p65,TLR4,NLRP3,Caspase-1蛋白表达升高,p65和TLR4 mRNA的表达升高(P<0.05);与模型组比较,喘可治组小鼠血清TNF-α,IL~(-1)β,IL~(-1)8含量降低(P<0.01),肺组织p65,TLR4,NLRP3,Caspase-1蛋白表达降低,p65和TLR4 mRNA的表达降低(P<0.05)。结论:喘可治注射液能有效改善LPS诱导的急性肺损伤小鼠肺组织的损伤程度,其机制可能通过抑制TLR4/NF-κB/NLRP3信号通路进而对肺组织起到保护作用。
Objective: To study the preventive and therapeutic effects and the mechanism of Chuankezhi injection on lipopolysaccharide (LPS) -induced acute lung injury (ALI) in mice. Methods: Forty-eight SPF female C57BL / 6J mice were randomly divided into normal group, model group, dexamethasone group and chuankezhi group, with 12 rats in each group. The mice in the asthma group were intraperitoneally injected with 0.2 m L · d-1, once per day for 7 days. The dexamethasone group was injected intraperitoneally with 1 mg · kg -1 Times / d, for 3 consecutive days. The model group was injected with normal saline without any treatment in the normal group. In addition to the normal group, LPS 0.1 μL (0.5 g · L -1) was intraperitoneally injected on the 8th day, and 25 μg LPS (0.5 g · L -1) Make an ALI model. After 24 h material testing. The levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and IL-1 8 in serum were detected by enzyme-linked immunosorbent assay The expression of nuclear factor-κB (NF-κB) p65 was detected by chemical staining. The expressions of p65, TLR4 and NLRP3 in lung tissue were detected by Western blot. Caspase-1 protein expression was detected by real-time PCR. The mRNA expression of p65 and TLR4 in lung tissue was detected by real-time PCR. Results: Compared with the normal group, the levels of TNF-α, IL-1β and IL-8 in the model group were significantly increased (P <0.01) and the expressions of p65, TLR4, NLRP3 and Caspase- (P <0.05). Compared with the model group, the levels of serum TNF-α, IL-1β, IL-1, 8 decreased (P <0.01). The expression of p65, TLR4, NLRP3 and Caspase-1 in lung tissue decreased and the expressions of p65 and TLR4 mRNA decreased (P <0.05). Conclusion: Chuankezhi Injection can effectively improve the lung injury induced by LPS in mice with acute lung injury. The mechanism may be through the inhibition of TLR4 / NF-κB / NLRP3 signaling pathway in lung tissue protection.