益气活血通络解毒方经CHOP通路对肺缺血/再灌注损伤的干预

YHTJF prevents lung ischemia-reperfusion injury via CHOP pathway

目的:研究益气活血通络解毒方(yiqi huoxue tongluo jiedu fang,YHTJF)减轻小鼠肺缺血/再灌注性损伤药效及其机制。方法:取雄性C57BL/6J小鼠70只,随机分为7组:正常对照组(control,C),羧甲基纤维素钠对照组(CMC-Na+control,CC),羧甲基纤维素钠假手术组(CMC-Na+sham,CS),羧甲基纤维素钠缺血/再灌注模型组(CMC-Na+I/R,CIR),YHTJF低、中、高浓度干预组(CMC-Na+YHTJF-low、CMC-Na+YHTJFmiddle、CMC-Na+YHTJF-high,CYL、CYM、CYH)。CC、CS、CIR组术前连续7 d腹腔注射CMC溶液,中药干预组术前连续7 d腹腔注射低、中、高浓度的YHTJF溶液。术毕,取左肺,测定肺组织干湿比(W/D)及总肺含水量(TLW),行肺组织损伤评估(IQA),光、电镜观察肺组织形态学及超微结构改变,原位末端标记法(TUNEL)测细胞凋亡指数(AI),Western blot和逆转录PCR(RT-PCR)分别检测CHOP、GRP78的蛋白和mRNA表达量。结果:较之C组,CIR组W/D、TLW、IQA、AI明显升高(P<0.01),肺组织形态学破坏显著,CHOP、GRP78蛋白和mRNA表达量增加(P<0.01),而CC、CS组各项指标与C组无明显差异;较CIR组,CYL、CYM、CYH中药保护剂组各项指标数值下降(P<0.01),组织损伤明显减轻,CYM组各项指标数值下降最明显,组织损伤最轻。结论:YHTJF可有效减轻小鼠缺血/再灌注性肺损伤,中剂量效果较好,该机制可能与其抑制CHOP通路所致凋亡有关。

AIM： To explore the principle and mechanism of yiqi huoxue tongluo jiedu fang （YHTJF） which can decrease the injury during lung ischemia-repeffusion （I/R） in mice. M^ETHODS ： 70 C57BL/6J male mice were randomly divided into 7 groups： control （C）, carboxyl methyl cellulose- Na-control （CC）, carboxyl methyl cellulose-Na- sham （ CS ） , carboxyl methyl eellulose-Na-I/R （ CIR ） , CMC-Na-YHTJF-low, CMC-Na-YHTJF- middle, CMC-Na-YHTJF-high （ CYL, CYM, CYH）. YHTJF was infused intraperitoneally with three different concentrations everyday in CYL, CYM, CYH groups, respectively, and the carboxyl methyl cellulose-Na was administrated with the same volume of CYL in CC, CS, CIR groups. After the 3 h reperfusion period finished, the left lung tissues were harvested to determine the lung wet/dry weight （W/D）, the total lung water content （TLW）, and index of quantitative evaluation for alveolar damage （IQA）. Morphological assay and Terminal-deoxynu- cleoitidyl Transferase Mediated Nick End Labeling （TUNEL） were applied to evaluate structure chan- ges and the apoptosis index （AI） of the lung tis- sues. The protein and mRNA expression of CHOP and GRP78 in lung tissues were detected by western blot and RT-PCR. RESULTS：Compared with group C, the W/D, TLW, IQA, AI, lung tissue structure changes, and the protein and mRNA expression of CHOP and grp78 in group I/R obviously raised （P 〈 0.01 ） , while there were no significant difference in group CC and CS. Compared with group I/R, the indexes of these experiments in group CYL, CYM, CYH all decreased, and the reduction in group CYM was the most remarkable among them （P 〈 0.01 ）. CONCLUSION： YHTJF may alleviate the LeR injury of lung, which may be related to inhibition of apoptosis via CHOP pathway.