广东医学
廣東醫學
엄동의학
GUNAGDONG MEDICAL JOURNAL
2014年
11期
1642-1646
,共5页
Toll 样受体 4%肿瘤坏死因子受体相关因子 6%白细胞介素 1 受体相关激酶 4%核转录因子 - κB
Toll 樣受體 4%腫瘤壞死因子受體相關因子 6%白細胞介素 1 受體相關激酶 4%覈轉錄因子 - κB
Toll 양수체 4%종류배사인자수체상관인자 6%백세포개소 1 수체상관격매 4%핵전록인자 - κB
Toll - like receptor 4%tumor necrosis factor receptor - associated factor 6%interleukin - 1 receptor -associated kinase 4%nuclear factor - κB
目的：探讨 Toll 样受体4（TLR4）及其相关信号链酶在不同潮气量机械通气大鼠肺组织中的表达变化。方法按随机数字表法将40只 SD 大鼠分为5组，每组8只。A 组仅切开气管保留自主呼吸，B1组和 B2组以潮气量6 mL/ kg 分别通气1 h 和2 h，C1组和 C2组以潮气量30 mL/ kg 分别通气1 h 和2 h。A 组在气管切开即刻，其余各组在机械通气结束时处死大鼠。光镜下观察各组大鼠肺组织病理改变，免疫组化法检测肺组织中TLR4、肿瘤坏死因子受体相关因子6（TRAF6）、白细胞介素1受体相关激酶4（IRAK4）及核转录因子-κB（NF -κB）p65蛋白的表达，RT - PCR 检测 TLR4 mRNA 的表达，测定肺组织髓过氧化物酶（MPO）的活性，计算肺湿干重比值（W/ D 比值），ELISA 法测定支气管肺泡灌洗液中肿瘤坏死因子-α（TNF -α）的浓度。结果 A 组、B1组、B2组之间肺组织 TLR4 mRNA 表达，TLR4、TRAF6、IRAK4、NF -κBp65蛋白的表达，W/ D 比值，MPO 活性和 TNF -α浓度差异均无统计学意义（P >0.05）。与相应 B1、B2组比较，C1、C2组 TLR4 mRNA 和 TLR4、TRAF6、IRAK4、NF -κBp65蛋白表达明显上调，W/ D 比值、MPO 活性和 TNF -α浓度明显升高，差异均有统计学意义（P <0.05）。随通气时间延长，C2组各指标较 C1组改变更为明显（P <0.05）。结论 TLR4及其相关信号链酶在肺组织中的表达参与了机械通气相关性肺损伤的发生与发展。
目的：探討 Toll 樣受體4（TLR4）及其相關信號鏈酶在不同潮氣量機械通氣大鼠肺組織中的錶達變化。方法按隨機數字錶法將40隻 SD 大鼠分為5組，每組8隻。A 組僅切開氣管保留自主呼吸，B1組和 B2組以潮氣量6 mL/ kg 分彆通氣1 h 和2 h，C1組和 C2組以潮氣量30 mL/ kg 分彆通氣1 h 和2 h。A 組在氣管切開即刻，其餘各組在機械通氣結束時處死大鼠。光鏡下觀察各組大鼠肺組織病理改變，免疫組化法檢測肺組織中TLR4、腫瘤壞死因子受體相關因子6（TRAF6）、白細胞介素1受體相關激酶4（IRAK4）及覈轉錄因子-κB（NF -κB）p65蛋白的錶達，RT - PCR 檢測 TLR4 mRNA 的錶達，測定肺組織髓過氧化物酶（MPO）的活性，計算肺濕榦重比值（W/ D 比值），ELISA 法測定支氣管肺泡灌洗液中腫瘤壞死因子-α（TNF -α）的濃度。結果 A 組、B1組、B2組之間肺組織 TLR4 mRNA 錶達，TLR4、TRAF6、IRAK4、NF -κBp65蛋白的錶達，W/ D 比值，MPO 活性和 TNF -α濃度差異均無統計學意義（P >0.05）。與相應 B1、B2組比較，C1、C2組 TLR4 mRNA 和 TLR4、TRAF6、IRAK4、NF -κBp65蛋白錶達明顯上調，W/ D 比值、MPO 活性和 TNF -α濃度明顯升高，差異均有統計學意義（P <0.05）。隨通氣時間延長，C2組各指標較 C1組改變更為明顯（P <0.05）。結論 TLR4及其相關信號鏈酶在肺組織中的錶達參與瞭機械通氣相關性肺損傷的髮生與髮展。
목적：탐토 Toll 양수체4（TLR4）급기상관신호련매재불동조기량궤계통기대서폐조직중적표체변화。방법안수궤수자표법장40지 SD 대서분위5조，매조8지。A 조부절개기관보류자주호흡，B1조화 B2조이조기량6 mL/ kg 분별통기1 h 화2 h，C1조화 C2조이조기량30 mL/ kg 분별통기1 h 화2 h。A 조재기관절개즉각，기여각조재궤계통기결속시처사대서。광경하관찰각조대서폐조직병리개변，면역조화법검측폐조직중TLR4、종류배사인자수체상관인자6（TRAF6）、백세포개소1수체상관격매4（IRAK4）급핵전록인자-κB（NF -κB）p65단백적표체，RT - PCR 검측 TLR4 mRNA 적표체，측정폐조직수과양화물매（MPO）적활성，계산폐습간중비치（W/ D 비치），ELISA 법측정지기관폐포관세액중종류배사인자-α（TNF -α）적농도。결과 A 조、B1조、B2조지간폐조직 TLR4 mRNA 표체，TLR4、TRAF6、IRAK4、NF -κBp65단백적표체，W/ D 비치，MPO 활성화 TNF -α농도차이균무통계학의의（P >0.05）。여상응 B1、B2조비교，C1、C2조 TLR4 mRNA 화 TLR4、TRAF6、IRAK4、NF -κBp65단백표체명현상조，W/ D 비치、MPO 활성화 TNF -α농도명현승고，차이균유통계학의의（P <0.05）。수통기시간연장，C2조각지표교 C1조개변경위명현（P <0.05）。결론 TLR4급기상관신호련매재폐조직중적표체삼여료궤계통기상관성폐손상적발생여발전。
Objective To investigate the expression of Toll - like receptor 4(TLR4)under different ventilator conditions and to explain the relationship between the expressions of TLR4 and the activation of downstream signals in lung tissue in stress injury model. Methods Forty healthy male SD rats were randomly assigned into 5 groups( n = 8):Group A(spontaneous breathing),Group B1 and B2(VT at 6 mL/ kg,durations of 1 h and 2 h,respectively),and Group C1 and C2(VT at 30 mL/ kg,durations of 1 h or 2 h,respectively). After maintaining ventilation for 1 h or 2 h, all rats were sacrificed and specimens of lung tissues and bronchoalveolar lavage fluid(BALF)were harvested. Lung path-ological changes were observed by microscopy. The protein levels of TLR4,tumor necrosis factor receptor - associated fac-tor 6(TRAF6),interleukin - 1 receptor - associated kinase 4(IRAK4)and nuclear factor - kappaB p65(NF - κBp65) in lung tissues were assayed with immunohistochemistry staining. The expression levels of TLR4 mRNA in lung tissue were measured by RT - PCR. Lung myeloperoxidase(MPO)activity was measured by colorimetric analysis,and wet - dry weight ratio(W/ D ratio)was also measured. The levels of tumor necrosis factor - α(TNF - α)in BALF were measured by ELISA. Results There was no significant difference in the expression of TLR4 mRNA and TLR4,TRAF6,IRAK4 and NF - κBp65 proteins,as well as W/ D ratio,MPO and TNF - α in BALF among Group A,Group B1 and Group B2 (all P > 0. 05). The expression of TLR4 mRNA and TLR4,TRAF6,IRAK4,NF - κBp65 proteins were significantly up- regulated,while W/ D ratio,MPO and TNF - α in BALF were also significantly increased in Group C1 and Group C2 than in Group B1 and Group B2 accordingly(all P < 0. 05). Each index changes were significantly more prominent in Group C2 than Group C1(all P < 0. 05). Conclusion TLR4 and the activation of the downstream signals in lung tissue participate in the development of ventilator - induced lung injury.
