β-羟丁酸对牛肺泡巨噬细胞促炎细胞因子释放的影响研究

β-羟丁酸(β-hydroxybutyrate,BHBA)除了能作为一种能源物质外,还可作为信号分子诱导肝细胞、子宫内膜细胞发生炎症反应,但其是否也能促使牛肺泡巨噬细胞(bovine alveolar macrophages, BAMs)发生炎症反应尚不清楚。利用不同浓度BHBA分别与 BAMs作用12、24 h,CCK-8法检测BAMs的存活率;用终浓度为4 mmol·L^-1 BHBA与 BAMs作用0、3、6、12、24 h,以及用0、1、2、4 mmol·L^-1 BHBA分别与 BAMs作用 12 h,qRT-PCR检测GPR109A、p38MAPK、NF-κB、PPARγ mRNA表达量,ELISA检测细胞上清中1L-1β、IL-6、TNF-α等的浓度。结果表明,1、2、4 mmol·L^-1 BHBA作用对BAMs 的存活率没有不良影响;在一定时间浓度范围内,GPR109A、p38MAPK和NF-κB mRNA表达量,1L-1β、IL-6、TNF-α等浓度随时间-剂量依赖变化(P<0.01),PPARγ mRNA表达量在3、6 h极显著上调(P<0.01),随后下调,PPARγ mRNA表达量只有2 mmol·L^-1组有显著变化(P<0.05)。综上,BHBA能促进BAMs促炎细胞因子的分泌和释放,导致其发生炎症反应。

Effect of beta-hydroxybutyrate on proinflammatory cytokine production in bovine alveolar macrophages

In addition to being an energy source, β-hydroxybutyrate (BHBA) can also be used as a signal molecule to induce inflammation in hepatocytes and endometrial cells. However, it is not clear whether BHBA can also induce inflammation in bovine alveolar macrophages (BAMs). BAMs were treated with different concentrations of BHBA for 12 and 24 h, respectively. The survival rate of BAMs was measured by CCK-8 method. BAMs were treated with 4 mmol·L ^-1 BHBA for 0, 3, 6, 12, 24 h and 0, 1, 2, 4 mmol·L^-1 BHBA for 12 h. The mRNA expressions of GPR109A, p38MAPK, NF-κB and PPARγ were detected by qRT-PCR, and the concentrations of 1L-1β, IL-6 and TNF-α in the supernatant were detected by ELISA. The 1, 2 and 4 mmol·L ^-1 BHBA had no adverse effect on the survival rate of BAMs. With in a certain time and concentration range, the mRNA expression of GPR109A, p38MAPK and NF-κB, and the concentrations of 1L-1β, IL-6 and TNF-α were time and dose-dependent (P<0.01). However, PPARγ mRNA expression was significantly increased at 3 and 6 h (P<0.01), and then down regulated. PPARγ mRNA expression was only significantly changed in 2 mmol·L ^-1 group (P<0.05). In conclusion, BHBA can promote the secretion and release of proinflammatory cytokines in BAMs, leading to the occurrence of inflammation.