益气活血通络解毒方对肺缺血／再灌注损伤小鼠细胞凋亡及c—Jun氨基末端激酶通路的影响  

Effects of traditional Chinese medicine Yiqi Huoxue Tongluo Jiedu prescription on pneumocyte apoptosis and c-Jun N-terminal protein kinase pathway in mice after lung ischemia/reperfusion injury

作　　者：石璐[1] 宋冬[3] 贾旭广[2] 罗梓垠 项冰倩 戴雍月[3] 罗岷[1] 王万铁[3] Shi Lu Song Dong Jia Xuguang Luo Ziyin Xiang Bingqian Dai Yongyue Luo Min Wang Wantie(Department of Physiology, Yibin Health School, Yibin 644000, Sichuan, China Department of Internal Medicine, Yibin Health School, Yibin 644000, Sichuan, China Ischemia/Reperfusion Injury Research Institute, Wenzhou Medical University, Wenzhou 325035, Zhejiang, China)

机构地区：[1]四川省宜宾卫生学校生理教研室,四川宜宾644000 [2]四川省宜宾卫生学校内科教研室,四川宜宾644000 [3]温州医科大学缺血/再灌注损伤研究所,浙江温州325035

出　　处：《中国中西医结合急救杂志》2017年第1期73-77,共5页Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care

基　　金：浙江省中医药重点研究计划（2013ZZ011）;四川省宜宾市重点科技计划项目（2015SF035）

摘　　要：目的观察益气活血通络解毒方（YHTJF）对小鼠肺缺血／再灌注（I／R）损伤（LIRI）的影响，并探讨c-Jun氨基末端激酶（JNK）是否参与其凋亡机制。方法选择雄性C57BL／6J小鼠70只，按随机数字表法分为正常对照组（C组）、羧甲基纤维素钠（CMC-Na）＋正常对照组（CMC-Na＋C组）、CMC-Na＋假手术组（CMC-Na＋S组）、CMC-Na＋I/R模型组和CMC-Na＋YHTJF低、中、高浓度干预组（CMC-Na＋YL组、CMC-Na＋YM组、CMC-Na＋YH组）。C组不进行任何处理；CMC-Na＋S组只开胸，不夹闭肺门；其余各组均开胸夹闭肺门30min，再松开动脉夹使左肺再灌注3h。术毕处死小鼠留取肺组织，光镜和电镜下观察肺组织形态学及超微结构改变，并观察肺泡损伤定量评估指标（IQA）的变化；用原位末端缺刻标记试验（TUNEL）检测肺组织细胞凋亡指数（AI）；用蛋白质免疫印迹试验（Western Blot）和反转录-聚酶链反应（RT-PCR）检测JNK、葡萄糖调节蛋白78（GRP78）的mRNA和蛋白表达；并分析肺组织AI与JNK、GRP78的mRNA和蛋白表达及IQA的相关性。结果CMC-Na＋I／R组IQA、AI及JNK和GRP78 mRNA和蛋白表达均较CMC-Na＋S组明显升高[IQA：（74．00±7．31）％比（7．00±1．23）％，AI：（64．40±11．97）％比（5．60±1．14）％，JNKmRNA（灰度值）：1．143±0．284比0．152±0．128，GRP78 mRNA（灰度值）：0．897±0．129比0．284±0．044，JNK蛋白（A值）：0．428±0．074比0．073±0．052，GRP78蛋白（A值）：1．075±0．145比0．589±0．060j，CMC-Na＋YL组、CMC-Na＋YM组、CMC-Na＋YH组IQA、AI、JNK mRNA和蛋白、GRP78mRNA表达均较CMC-Na＋I／R组明显下降，以CMC-Na＋YM组较CMC-Na＋YL和CMC-Na＋YH组下降程度更显著[IQA：（26．20±3．35）％比（34．00±5．34）％、（41．20±9．18）％，AI：（29．40±3．05）％比（48．20±3．83）％、（39．20±6．14）％，JNK mRNA（灰度值）：0．681±0．Objective To observe the effects of Yiqi Huoxue Tongluo Jiedu fang （YHTJF） on pneumocyte apoptosis after lung isehemia/reperfusion （I/R） injury （LIRI） in mice and to investigate whether c-Jun N-terminal protein kinase （,INK） is involved in the mechanism of apoptosis. Methods Seventy C57BL/6J male mice were randomly divided into seven groups： normal control group （C group）, carboxyl methyl cellulose-Na＋ormal control group （CMC-Na＋ group）, CMC-Na＋ham group （CMC-Na＋ group）, CMC-Na＋/R group （CMC-Na＋/R group） and CMC-Na＋HTJF-low, -middle, -high dose groups （CMC-Na＋L, CMC-Na＋M, CMC-Na＋H groups）. C group did not undergo any processing; in CMC-Na＋ group, only was chest opened without clipping the lung hilum; in the rest of the four groups, they all underwent opening of the chest and clipping the lung hilum for 30 minutes, then the clipping of artery was relieved and left lung repeffusion was carried out for 3 hours. After operation, the mice were sacrificed, the lung tissues were harvested. Under light and electron microscopes, the lung morphological and ultra-structural changes were observed, and the changes of index of quantitative evaluation for alveolar damage （IQA） were determined. The terminal-deoxynucleoitidyl transferase mediated nick end labeling （TUNEL） was applied to evaluate the apoptosis index （AI） of the lung tissues. The protein and mRNA expressions of JNK and glucose regulating protein 78 （GRP78） in lung tissues were detected by Western Blot and reverse transcription-polymerase chain reaction （RT-PCR）; the correlations between lung AI and the expressions of mRNA and protein of JNK and GRP78, IQA were analyzed. Results Compared with CMC-Na＋ group, IQA, AI and mRNA and the protein expressions of JNK and GRP78 in CMC-Na＋/R group were obviously higher [IQA： （74.00 ± 7.31）% vs. （7.00± 1.23）%, AI： （64.40 ± 11.97）% vs. （5.60 ± 1.14）%, JNK mRNA （gray value）： 1.143 ± 0.284 vs. 0.152 ± 0.128, GRP78 mR

关 键 词：益气活血通络解毒方 c—Jun氨基末端激酶 缺血/再灌注损伤 肺 细胞凋亡 小鼠 

分 类 号：R978.1[医药卫生—药品]