中国危重病急救医学
中國危重病急救醫學
중국위중병급구의학
CHINESE CRITICAL CARE MEDICINE
2012年
3期
166-169
,共4页
曾振国%龚洪翰%李勇%聂贞蕴%揭克敏%詹以安%聂成%刘芬%丁成志%邵强%卿城%朱白鹭%钱克俭
曾振國%龔洪翰%李勇%聶貞蘊%揭剋敏%詹以安%聶成%劉芬%丁成誌%邵彊%卿城%硃白鷺%錢剋儉
증진국%공홍한%리용%섭정온%게극민%첨이안%섭성%류분%정성지%소강%경성%주백로%전극검
参附注射液%微小RNA-146a%肿瘤坏死因子-α%肺泡巨噬细胞
參附註射液%微小RNA-146a%腫瘤壞死因子-α%肺泡巨噬細胞
삼부주사액%미소RNA-146a%종류배사인자-α%폐포거서세포
Shenfu injection%MicroRNA-146a%Tumor necrosis factor-α%Alveolar macrophage
目的 观察参附注射液(SF)对脂多糖(LPS)诱导肺泡巨噬细胞微小RNA-146a(miR-146a)表达的影响以及SF的可能抗炎机制.方法 将体外培养的大鼠NR8383肺泡巨噬细胞分为对照组、LPS刺激组、SF处理组.LPS刺激组培养基中LPS终浓度为1 mg/L,对照组给予LPS刺激组等体积的磷酸盐缓冲液(PBS)；SF处理组细胞分别用不同浓度的SF(1 ml/L或10 ml/L)预处理30 min后,再加入LPS(终浓度1 mg/L).给予PBS或LPS刺激6h后收集细胞和上清液,检测各组细胞中miR-146a表达水平(逆转录-聚合酶链反应法)和上清液中肿瘤坏死因子-α(TNF-α)含量(酶联免疫吸附试验).结果 与对照组相比,LPS刺激组细胞中miR-146a表达和上清液中TNF-α含量显著增高[miR-146a(表达倍数):5.92±1.57比1.04±0.38；TNF-α(ng/L):636.93±30.21比20.46±2.81;均P＜0.05].与LPS刺激组相比,1ml/L和10 ml/L的SF预处理后,细胞中miR-146a表达均显著增高,而上清液中TNF-α含量均显著下降[miR-146a(表达倍数):7.02±0.91、8.11±1.07比5.92±1.57; TN F-α(ng/L):447.24±21.29、357.83±19.73比636.93±30.21,均P＜ 0.05],且均呈剂量依赖性(均P＜0.05).结论 SF可以呈剂量依赖性地上调肺泡巨噬细胞miR-146a的表达水平,推测miR-146a参与了SF的抗炎过程.
目的 觀察參附註射液(SF)對脂多糖(LPS)誘導肺泡巨噬細胞微小RNA-146a(miR-146a)錶達的影響以及SF的可能抗炎機製.方法 將體外培養的大鼠NR8383肺泡巨噬細胞分為對照組、LPS刺激組、SF處理組.LPS刺激組培養基中LPS終濃度為1 mg/L,對照組給予LPS刺激組等體積的燐痠鹽緩遲液(PBS)；SF處理組細胞分彆用不同濃度的SF(1 ml/L或10 ml/L)預處理30 min後,再加入LPS(終濃度1 mg/L).給予PBS或LPS刺激6h後收集細胞和上清液,檢測各組細胞中miR-146a錶達水平(逆轉錄-聚閤酶鏈反應法)和上清液中腫瘤壞死因子-α(TNF-α)含量(酶聯免疫吸附試驗).結果 與對照組相比,LPS刺激組細胞中miR-146a錶達和上清液中TNF-α含量顯著增高[miR-146a(錶達倍數):5.92±1.57比1.04±0.38；TNF-α(ng/L):636.93±30.21比20.46±2.81;均P＜0.05].與LPS刺激組相比,1ml/L和10 ml/L的SF預處理後,細胞中miR-146a錶達均顯著增高,而上清液中TNF-α含量均顯著下降[miR-146a(錶達倍數):7.02±0.91、8.11±1.07比5.92±1.57; TN F-α(ng/L):447.24±21.29、357.83±19.73比636.93±30.21,均P＜ 0.05],且均呈劑量依賴性(均P＜0.05).結論 SF可以呈劑量依賴性地上調肺泡巨噬細胞miR-146a的錶達水平,推測miR-146a參與瞭SF的抗炎過程.
목적 관찰삼부주사액(SF)대지다당(LPS)유도폐포거서세포미소RNA-146a(miR-146a)표체적영향이급SF적가능항염궤제.방법 장체외배양적대서NR8383폐포거서세포분위대조조、LPS자격조、SF처리조.LPS자격조배양기중LPS종농도위1 mg/L,대조조급여LPS자격조등체적적린산염완충액(PBS)；SF처리조세포분별용불동농도적SF(1 ml/L혹10 ml/L)예처리30 min후,재가입LPS(종농도1 mg/L).급여PBS혹LPS자격6h후수집세포화상청액,검측각조세포중miR-146a표체수평(역전록-취합매련반응법)화상청액중종류배사인자-α(TNF-α)함량(매련면역흡부시험).결과 여대조조상비,LPS자격조세포중miR-146a표체화상청액중TNF-α함량현저증고[miR-146a(표체배수):5.92±1.57비1.04±0.38；TNF-α(ng/L):636.93±30.21비20.46±2.81;균P＜0.05].여LPS자격조상비,1ml/L화10 ml/L적SF예처리후,세포중miR-146a표체균현저증고,이상청액중TNF-α함량균현저하강[miR-146a(표체배수):7.02±0.91、8.11±1.07비5.92±1.57; TN F-α(ng/L):447.24±21.29、357.83±19.73비636.93±30.21,균P＜ 0.05],차균정제량의뢰성(균P＜0.05).결론 SF가이정제량의뢰성지상조폐포거서세포miR-146a적표체수평,추측miR-146a삼여료SF적항염과정.
Objective To study the effects of Shenfu injection(SF)on the expression of lipopolysaccharide (LPS)-induced microRNA-146a(miR-146a)in rat alveolar macrophages(AMs),and to extrapolate its potential anti-inflammatory mechanisms.Methods In vitro cultured rat AMs(NR8383 cells)were randomly divided into control group,LPS stimulation group,and SF stimulation group.The LPS stimulation group was challenged with a final concentration of 1 mg/L LPS,and to the control group an equal volume of phosphate buffer solution(PBS)was added instead.For SF treated group,SF in different concentrations(1 ml/L or 10 ml/L)was used during incubation of AMs for half an hour,and then LPS was added(1 mg/L final concentration).After 6 hours,the cells and culture supematants were collected.MiRNA-146a expression[reverse transcription-polymerase chain reaction(RT-PCR)]in cells and tumor necrosis factor-α(TNF-α)content[enzyme-linked immunosorbent assay(ELISA)]in culture supernatant were determined for each group.Results Both the expression of miR-146a and TNF-α content in LPS stimulation group were significantly elevated compared with control group[miR-146a(expression folds):5.92 ± 1.57 vs.1.04 ±0.38; TNF-α(ng/L):636.93 ± 30.21 vs.20.46 ± 2.81 ; both P＜0.05].Compared with LPS stimulation group,the expression of miR-146a was significantly upregulated in cells in both 1 ml/L and 10 ml/L SF stimulation groups,but TNF-α content was significantly reduced in the supernatant[miR-146a(expression folds):7.02 ± 0.91,8.11 ± 1.07vs.5.92 ±1.57; TNF-α(ng/L):447.24 ±21.29,357.83 ±19.73 vs.636.93 ±30.21,all P＜0.05]in a dose-dependent manner(both P＜0.05).Conclusion SF could up-regulate miR-146a expression in AMs in a dose-dependent manner,and it was speculated that miR-146a might be involved in the anti-inflammatory processes with SF treatment.
