中国中西医结合急救杂志
中國中西醫結閤急救雜誌
중국중서의결합급구잡지
INTEGRATED TRADITIONAL CHINESE AND WESTERN MEDICINE IN PRACTICE OF CRITICAL CARE MEDICINE
2015年
1期
23-27
,共5页
刘国跃%陈淼%戢慧%覃松%梅鸿%陈涛%陈华军
劉國躍%陳淼%戢慧%覃鬆%梅鴻%陳濤%陳華軍
류국약%진묘%집혜%담송%매홍%진도%진화군
微小RNA-21-5p%肺损伤,急性,高氧性%氧合指数%呼吸指数%肺湿/干质量比值
微小RNA-21-5p%肺損傷,急性,高氧性%氧閤指數%呼吸指數%肺濕/榦質量比值
미소RNA-21-5p%폐손상,급성,고양성%양합지수%호흡지수%폐습/간질량비치
miRNA-21-5p%Hyperoxic acute lung injury%Oxygenation index%Respiratory index%Lung wet/dry weight ratio
目的：探讨微小RNA-21-5p（miR-21-5p）对大鼠高氧性急性肺损伤（HALI）的影响，为HALI的基因治疗提供理论依据。方法按随机数字表法将160只SD大鼠分为高氧对照组、磷酸盐缓冲液（PBS）组、空病毒组及miR-21-5p组4组，每组40只。高氧对照组大鼠直接置于高氧箱（氧浓度＞90%）中饲养，其余3组分别经鼻腔滴入PBS、慢病毒及miR-21-5p慢病毒各200μL，然后置于高氧箱中饲养。各组分别在高氧放置0、24、48及72 h时随机取10只大鼠进行动脉血气分析，并计算氧合指数（OI）和呼吸指数（RI）；随后颈动脉放血处死大鼠取肺组织，测量左肺湿/干质量（W/D）比值，行苏木素-伊红（HE）染色，光镜下观察右肺组织病理学改变并进行病理学评分。结果高氧损伤0 h时，各组大鼠OI、RI、肺组织病理学评分、肺W/D比值差异均无统计学意义（均P＞0.05）；随着高氧损伤时间的延长，各组OI逐渐降低，RI、肺组织病理学评分、肺W/D比值逐渐升高。与高氧对照组比较，miR-21-5p组高氧损伤24、48、72 h时OI明显增加〔mmHg（1 mmHg＝0.133 kPa）：24 h 时358.10±29.25比306.19±37.23，48 h 时336.67±29.27比269.70±29.00，72 h 时323.81±19.05比203.81±43.40，均P＜0.05〕，RI明显减少（24 h时0.23±0.05比0.31±0.06，48 h时0.28±0.07比0.38±0.06，72 h时0.30±0.04比0.46±0.07，均P＜0.05），肺组织病理学评分明显降低（分：24 h时0.60±0.52比0.90±0.74，48 h时1.30±0.95比2.90±1.20，72 h时1.90±0.88比4.70±1.57，均P＜0.05），肺W/D比值明显下降（24 h时3.77±0.38比4.14±0.46，48 h时3.83±0.31比4.56±0.34，72 h时3.89±0.31比5.32±0.27，均P＜0.05）。PBS组、空病毒组高氧损伤后各指标与高氧对照组比较差异均无统计学意义（均P＞0.05）。光镜下观察可见，随着高氧损伤时间的延长，各组大鼠肺泡结构逐渐紊乱，肺泡壁断裂破坏，肺泡间隔逐渐增宽、水肿、炎性细胞浸润，部分大鼠可见少量红细胞渗出；但miR-21-5p组肺组织病理损伤程度较其他各组明显减轻。结论持续高浓度氧暴露24 h后可成功建立HALI模型；miR-21-5p对HALI大鼠肺组织具有一定的保护作用。
目的：探討微小RNA-21-5p（miR-21-5p）對大鼠高氧性急性肺損傷（HALI）的影響，為HALI的基因治療提供理論依據。方法按隨機數字錶法將160隻SD大鼠分為高氧對照組、燐痠鹽緩遲液（PBS）組、空病毒組及miR-21-5p組4組，每組40隻。高氧對照組大鼠直接置于高氧箱（氧濃度＞90%）中飼養，其餘3組分彆經鼻腔滴入PBS、慢病毒及miR-21-5p慢病毒各200μL，然後置于高氧箱中飼養。各組分彆在高氧放置0、24、48及72 h時隨機取10隻大鼠進行動脈血氣分析，併計算氧閤指數（OI）和呼吸指數（RI）；隨後頸動脈放血處死大鼠取肺組織，測量左肺濕/榦質量（W/D）比值，行囌木素-伊紅（HE）染色，光鏡下觀察右肺組織病理學改變併進行病理學評分。結果高氧損傷0 h時，各組大鼠OI、RI、肺組織病理學評分、肺W/D比值差異均無統計學意義（均P＞0.05）；隨著高氧損傷時間的延長，各組OI逐漸降低，RI、肺組織病理學評分、肺W/D比值逐漸升高。與高氧對照組比較，miR-21-5p組高氧損傷24、48、72 h時OI明顯增加〔mmHg（1 mmHg＝0.133 kPa）：24 h 時358.10±29.25比306.19±37.23，48 h 時336.67±29.27比269.70±29.00，72 h 時323.81±19.05比203.81±43.40，均P＜0.05〕，RI明顯減少（24 h時0.23±0.05比0.31±0.06，48 h時0.28±0.07比0.38±0.06，72 h時0.30±0.04比0.46±0.07，均P＜0.05），肺組織病理學評分明顯降低（分：24 h時0.60±0.52比0.90±0.74，48 h時1.30±0.95比2.90±1.20，72 h時1.90±0.88比4.70±1.57，均P＜0.05），肺W/D比值明顯下降（24 h時3.77±0.38比4.14±0.46，48 h時3.83±0.31比4.56±0.34，72 h時3.89±0.31比5.32±0.27，均P＜0.05）。PBS組、空病毒組高氧損傷後各指標與高氧對照組比較差異均無統計學意義（均P＞0.05）。光鏡下觀察可見，隨著高氧損傷時間的延長，各組大鼠肺泡結構逐漸紊亂，肺泡壁斷裂破壞，肺泡間隔逐漸增寬、水腫、炎性細胞浸潤，部分大鼠可見少量紅細胞滲齣；但miR-21-5p組肺組織病理損傷程度較其他各組明顯減輕。結論持續高濃度氧暴露24 h後可成功建立HALI模型；miR-21-5p對HALI大鼠肺組織具有一定的保護作用。
목적：탐토미소RNA-21-5p（miR-21-5p）대대서고양성급성폐손상（HALI）적영향，위HALI적기인치료제공이론의거。방법안수궤수자표법장160지SD대서분위고양대조조、린산염완충액（PBS）조、공병독조급miR-21-5p조4조，매조40지。고양대조조대서직접치우고양상（양농도＞90%）중사양，기여3조분별경비강적입PBS、만병독급miR-21-5p만병독각200μL，연후치우고양상중사양。각조분별재고양방치0、24、48급72 h시수궤취10지대서진행동맥혈기분석，병계산양합지수（OI）화호흡지수（RI）；수후경동맥방혈처사대서취폐조직，측량좌폐습/간질량（W/D）비치，행소목소-이홍（HE）염색，광경하관찰우폐조직병이학개변병진행병이학평분。결과고양손상0 h시，각조대서OI、RI、폐조직병이학평분、폐W/D비치차이균무통계학의의（균P＞0.05）；수착고양손상시간적연장，각조OI축점강저，RI、폐조직병이학평분、폐W/D비치축점승고。여고양대조조비교，miR-21-5p조고양손상24、48、72 h시OI명현증가〔mmHg（1 mmHg＝0.133 kPa）：24 h 시358.10±29.25비306.19±37.23，48 h 시336.67±29.27비269.70±29.00，72 h 시323.81±19.05비203.81±43.40，균P＜0.05〕，RI명현감소（24 h시0.23±0.05비0.31±0.06，48 h시0.28±0.07비0.38±0.06，72 h시0.30±0.04비0.46±0.07，균P＜0.05），폐조직병이학평분명현강저（분：24 h시0.60±0.52비0.90±0.74，48 h시1.30±0.95비2.90±1.20，72 h시1.90±0.88비4.70±1.57，균P＜0.05），폐W/D비치명현하강（24 h시3.77±0.38비4.14±0.46，48 h시3.83±0.31비4.56±0.34，72 h시3.89±0.31비5.32±0.27，균P＜0.05）。PBS조、공병독조고양손상후각지표여고양대조조비교차이균무통계학의의（균P＞0.05）。광경하관찰가견，수착고양손상시간적연장，각조대서폐포결구축점문란，폐포벽단렬파배，폐포간격축점증관、수종、염성세포침윤，부분대서가견소량홍세포삼출；단miR-21-5p조폐조직병리손상정도교기타각조명현감경。결론지속고농도양폭로24 h후가성공건립HALI모형；miR-21-5p대HALI대서폐조직구유일정적보호작용。
Objective To investigate the effects of microRNA-21-5p (miR-21-5p) on hyperoxic acute lung injury (HALI) in rats and provide a theoretical basis for HALI gene therapy. Methods One hundred and sixty Sprague-Dawley (SD) rats were randomly divided into four groups with number table:hyperoxia control group, phosphate buffer saline (PBS) group, blank virus group and miRNA-21-5p group (each, n = 40). The rats in hyperoxia control group were fed directly in the hyperoxia box (oxygen concentration > 90%); in the other three groups, 200 μL PBS, 200μL slow virus and 200μL miRNA-21-5p slow virus were dropped into the nose respectively, and then they were fed in the hyperoxia box. The rats were exposed to hyperoxia in the boxes for 0, 24, 48 and 72 hours in all the groups, and at each time point, 10 rats were taken randomly from each group to perform arterial blood-gas analysis, calculate oxygenation index (OI) and respiratory index (RI). Afterwards the rats were sacrificed by blood-letting from carotid artery under intra-peritoneal anesthesia, and the lung tissues were obtained to measure the left lung wet/dry weight (W/D) ratio, hemotoxylin-eosin (HE) staining was made and the pathological changes of the right lung were observed under light microscope and the pathological score was measured. Results At 0 hour, the OI, RI, lung W/D ratio and the lung tissue pathology score in rats with hyperoxic injury had no statistically significant differences among the four groups (all P>0.05). With the extension of time, the level of OI was gradually reduced, and the levels of RI, pathologic score and W/D ratio of lung tissues were gradually increased. Compared with the hyperoxia control group, in miRNA-21-5p group, the levels of OI were increased significantly at 24, 48 and 72 hours after the exposure to hyperoxia [mmHg (1 mmHg = 0.133 kPa): 24 hours 358.10±29.25 vs. 306.19±37.23, 48 hours 336.67±29.27 vs. 269.70±29.00, 72 hours 323.81±19.05 vs. 203.81±43.40, all P < 0.05], whereas the levels of RI were decreased significantly (24 hours 0.23±0.05 vs. 0.31±0.06, 48 hours 0.28±0.07 vs. 0.38±0.06, 72 hours 0.30±0.04 vs. 0.46±0.07, all P <0.05), the pathologic scores were decreased significantly (24 hours 0.60±0.52 vs. 0.90±0.74, 48 hours 1.30±0.95 vs.2.90±1.20, 72 hours 1.90±0.88 vs. 4.70±1.57, all P < 0.05) and the levels of W/D ratio were decreased obviously (24 hours 3.77±0.38 vs. 4.14±0.46, 48 hours 3.83±0.31 vs. 4.56±0.34, 72 hours 3.89±0.31 vs. 5.32±0.27, all P<0.05). Compared with the hyperoxia control group, the index results of the PBS group and the blank virus group after staying in the box had no statistical significant differences at each time point (all P>0.05). Under the optical microscope, along with the prolongation of exposure to hyperoxia, the structure of alveoli was gradually disturbed, their walls fractured and damaged, alveolar septa widened, edematous, infiltrated with inflammatory cells and in part of the rats a small amount of red blood cell exudates could be seen, but the degree of lung pathological injury in miRNA-21-5p group was much milder than that of the other groups. Conclusion The rat persistently exposed to hyperoxia for 24 hours can establish the rat model of HALI successfully, and the miRNA-21-5p can protect the lung tissue from the damage to some degrees in HALI rats.
