CAJ | 학술논문

目的：探讨血必净注射液对急性肺血栓栓塞症（APTE）的治疗作用及可能机制。方法选择纯种新西兰大耳白兔32只，按随机数字表法分为正常对照组、模型组、低分子肝素组、血必净组，每组8只。采用自体血栓回输法建立肺血栓栓塞模型；正常对照组不予任何处理。血必净组制模成功后给予血必净注射液静脉推注（静推）1 mL/kg；低分子肝素组给予低分子肝素钠100 U/kg皮下注射；模型组和正常对照组静注等量生理盐水；均每日2次，共7 d。采用酶联免疫吸附试验（ELISA）测定制模后8 h和1、3、5、7 d的血浆D-二聚体及内皮素-1（ET-1）含量变化；7 d后处死动物取肺组织，透射电镜下观察超微结构变化。结果模型组D-二聚体随制模时间的延长逐渐升高，制模后7 d明显高于正常对照组、低分子肝素组和血必净组（g/L：3.98±1.14比0.43±0.12、1.05±0.13、1.04±0.15，均P＜0.05）；ET-1于制模后5 d达峰值，制模后7 d有所降低，但仍明显高于正常对照组、低分子肝素组和血必净组（g/L：63.8±20.9比20.0±1.4、36.5±8.3、34.2±6.3，均P＜0.05）。低分子肝素组和血必净组各时间点D-二聚体、ET-1水平均明显高于正常对照组，但均明显低于模型组（均P＜0.05），两治疗组各时间点比较差异均无统计学意义（均P＞0.05）。电镜下观察肺组织超微结构显示：模型组肺组织基底膜断裂，血管内皮细胞，Ⅰ、Ⅱ型肺泡上皮细胞器结构不清晰，细胞膜表面微绒毛消失，细胞质浓缩，细胞核染色质固缩，可见大量核裂解碎片，脱落后形成凋亡小体，线粒体结构破坏；低分子肝素组毛细血管内皮细胞肿胀，可见大量坏死血细胞渗出，堵塞管腔，Ⅱ型肺泡上皮细胞胞质疏松，明显水肿，多数板层小体空泡化；血必净组毛细血管内皮细胞肿胀，管腔内可见坏死血细胞渗出，堵塞管腔，链接基本正常，Ⅱ型肺泡上皮细胞水肿，板层小体排空减轻。结论血必净能够显著降低APTE兔血浆D-二聚体、ET-1水平，调整凝血和纤溶平衡，同时使肺血管通透性显著降低，具有一定的内皮功能保护作用，减少肺损伤，减轻了肺栓塞后肺组织的病理损害。目的：探討血必淨註射液對急性肺血栓栓塞癥（APTE）的治療作用及可能機製。方法選擇純種新西蘭大耳白兔32隻，按隨機數字錶法分為正常對照組、模型組、低分子肝素組、血必淨組，每組8隻。採用自體血栓迴輸法建立肺血栓栓塞模型；正常對照組不予任何處理。血必淨組製模成功後給予血必淨註射液靜脈推註（靜推）1 mL/kg；低分子肝素組給予低分子肝素鈉100 U/kg皮下註射；模型組和正常對照組靜註等量生理鹽水；均每日2次，共7 d。採用酶聯免疫吸附試驗（ELISA）測定製模後8 h和1、3、5、7 d的血漿D-二聚體及內皮素-1（ET-1）含量變化；7 d後處死動物取肺組織，透射電鏡下觀察超微結構變化。結果模型組D-二聚體隨製模時間的延長逐漸升高，製模後7 d明顯高于正常對照組、低分子肝素組和血必淨組（g/L：3.98±1.14比0.43±0.12、1.05±0.13、1.04±0.15，均P＜0.05）；ET-1于製模後5 d達峰值，製模後7 d有所降低，但仍明顯高于正常對照組、低分子肝素組和血必淨組（g/L：63.8±20.9比20.0±1.4、36.5±8.3、34.2±6.3，均P＜0.05）。低分子肝素組和血必淨組各時間點D-二聚體、ET-1水平均明顯高于正常對照組，但均明顯低于模型組（均P＜0.05），兩治療組各時間點比較差異均無統計學意義（均P＞0.05）。電鏡下觀察肺組織超微結構顯示：模型組肺組織基底膜斷裂，血管內皮細胞，Ⅰ、Ⅱ型肺泡上皮細胞器結構不清晰，細胞膜錶麵微絨毛消失，細胞質濃縮，細胞覈染色質固縮，可見大量覈裂解碎片，脫落後形成凋亡小體，線粒體結構破壞；低分子肝素組毛細血管內皮細胞腫脹，可見大量壞死血細胞滲齣，堵塞管腔，Ⅱ型肺泡上皮細胞胞質疏鬆，明顯水腫，多數闆層小體空泡化；血必淨組毛細血管內皮細胞腫脹，管腔內可見壞死血細胞滲齣，堵塞管腔，鏈接基本正常，Ⅱ型肺泡上皮細胞水腫，闆層小體排空減輕。結論血必淨能夠顯著降低APTE兔血漿D-二聚體、ET-1水平，調整凝血和纖溶平衡，同時使肺血管通透性顯著降低，具有一定的內皮功能保護作用，減少肺損傷，減輕瞭肺栓塞後肺組織的病理損害。
목적：탐토혈필정주사액대급성폐혈전전새증（APTE）적치료작용급가능궤제。방법선택순충신서란대이백토32지，안수궤수자표법분위정상대조조、모형조、저분자간소조、혈필정조，매조8지。채용자체혈전회수법건립폐혈전전새모형；정상대조조불여임하처리。혈필정조제모성공후급여혈필정주사액정맥추주（정추）1 mL/kg；저분자간소조급여저분자간소납100 U/kg피하주사；모형조화정상대조조정주등량생리염수；균매일2차，공7 d。채용매련면역흡부시험（ELISA）측정제모후8 h화1、3、5、7 d적혈장D-이취체급내피소-1（ET-1）함량변화；7 d후처사동물취폐조직，투사전경하관찰초미결구변화。결과모형조D-이취체수제모시간적연장축점승고，제모후7 d명현고우정상대조조、저분자간소조화혈필정조（g/L：3.98±1.14비0.43±0.12、1.05±0.13、1.04±0.15，균P＜0.05）；ET-1우제모후5 d체봉치，제모후7 d유소강저，단잉명현고우정상대조조、저분자간소조화혈필정조（g/L：63.8±20.9비20.0±1.4、36.5±8.3、34.2±6.3，균P＜0.05）。저분자간소조화혈필정조각시간점D-이취체、ET-1수평균명현고우정상대조조，단균명현저우모형조（균P＜0.05），량치료조각시간점비교차이균무통계학의의（균P＞0.05）。전경하관찰폐조직초미결구현시：모형조폐조직기저막단렬，혈관내피세포，Ⅰ、Ⅱ형폐포상피세포기결구불청석，세포막표면미융모소실，세포질농축，세포핵염색질고축，가견대량핵렬해쇄편，탈락후형성조망소체，선립체결구파배；저분자간소조모세혈관내피세포종창，가견대량배사혈세포삼출，도새관강，Ⅱ형폐포상피세포포질소송，명현수종，다수판층소체공포화；혈필정조모세혈관내피세포종창，관강내가견배사혈세포삼출，도새관강，련접기본정상，Ⅱ형폐포상피세포수종，판층소체배공감경。결론혈필정능구현저강저APTE토혈장D-이취체、ET-1수평，조정응혈화섬용평형，동시사폐혈관통투성현저강저，구유일정적내피공능보호작용，감소폐손상，감경료폐전새후폐조직적병리손해。
Objective To explore the therapeutic effect and possible mechanisms of Xuebijing injection for treatment of rabbits with acute pulmonary thromboembolism (APTE). Methods Thirty-two New Zealand rabbits were randomly divided into normal control group, model group, low molecular weight heparin (LMWH) group and Xuebijing group by random number table, 8 rabbits in each group. APTE model was reproduced by re-infusing autologous blood clots, and nothing was done for the normal control group. After the model was established, intravenous injection of Xuebijing 1 mL/kg from ear marginal vein was given to the rabbits in Xuebijing group, the LMWH 100 U/kg was subcutaneously injected in LMWH group, the same amount of normal saline was given intravenously to model group and normal control group, twice daily for 7 days in all the groups. The changes of plasma D-dimer and endothelin-1 (ET-1) levels were detected by enzyme-linked immunosorbent assay (ELISA) after modeling for 8 hours and 1, 3, 5, 7 days;after 7 days the animals were sacrificed and their lung tissues were removed for the examination of ultrastructural changes under an electron microscope. Results With the prolongation of time, the D-dimer level in the model group was gradually increased, and on the 7th day after modeling, the level was significantly higher than that of the normal control, LMWH and Xuebijing groups (g/L:3.98±1.14 vs. 0.43±0.12, 1.05±0.13, 1.04±0.15, all P<0.05). After modeling for 5 days, the ET-1 level reached its peak in the model group, and it was lowered on the 7th day after modeling, but still significantly higher than that in the normal control, LMWH and Xuebijing groups (g/L:63.8±20.9 vs. 20.0±1.4, 36.5±8.3, 34.2±6.3, all P<0.05). The D-dimer and ET-1 levels in the LMWH and Xuebijing groups were significantly higher than those of the normal control group at each time point, but still obviously lower than those of the model group (all P<0.05), and the differences between the two treatment groups were not statistically significant at each time point (all P>0.05). Electron microscopic examination of lung tissue ultra-structure showed:in the model group, there were the lung tissue basement membrane rupture, the organelle structure in vascular endothelial cells and alveolar epithelial cells typeⅠandⅡnot clear, disappearance of microvilli on cell surface, cytoplasm concentration, nuclear chromatin condensation, a lot of debris due to nuclear lysis, formation of apoptotic bodies after shedding, and mitochondrial structural damage. In LMWH group, the capillary endothelial cell swelling, a lot of necrotized blood cells oozing, blockage of lumen, the cytoplasm of typeⅡalveolar epithelial cell loose, edematous and multiple lamellar bodies vacuolized were seen. In Xuebijing group, capillary endothelial cell swelling, necrotized blood cells blocking the lumen and seeping, the link basically normal, the type Ⅱ alveolar epithelial cell edematous, mitigation of lamellar bodies emptying were found. Conclusions Xuebijing can significantly reduce the plasma D-dimer and ET-1 levels, adjust the balance of coagulation and fibrinolysis, simultaneously decrease the pulmonary vascular permeability obviously in rabbits with APTE, thus it possesses certain protective effect for endothelial function leading to amelioration of lung injury and reduction of lung pathological damage in rabbits with pulmonary embolism.