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目的:探讨益气活血通络解毒方(Yiqi Huoxue Tongluo Jiedu Fang,YHTJF)对小鼠肺部缺血/再灌注损伤及caspase-12表达的影响。方法:选取雄性10~12周龄C57BL/6J小鼠70只,体重21~25g,复制在体左肺缺血/再灌注(ischemia-reperfusion,I/R)损伤模型。随机分为7组:正常对照(control)组,羧甲基纤维素钠(carboxyl methyl cellulose-Na,CMC-Na)组,假手术(sham)组,I/R模型组,YHTJF低(low,L)、中(meddle,M)、高(high,H)浓度干预组。CMC-Na、sham和I/R组术前连续7 d腹腔注射CMC-Na溶液,YHTJF-L、YHTJF-M和YHTJF-H组术前连续7 d腹腔注射低、中、高浓度的YHTJF溶液。术毕,取左肺测定肺组织湿/干比值(W/D)及总肺含水量(TLW),行肺泡损伤评估(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)测组织细胞凋亡指数(AI);RT-PCR和Western blot分别检测caspase-12和葡萄糖调节蛋白78(GRP78)的mRNA和蛋白表达量。结果:相比control组,I/R组的W/D、TLW、IQA和AI明显升高(P<0.01),肺组织形态学破坏显著,caspase-12和GRP78的mRNA和蛋白表达量增加(P<0.01),而CMC-Na组和sham组各项指标与control组无明显差异;与I/R组比较,YHTJF-L组、YHTJF-M组和YHTJF-H组各项指标(除GRP78外)数值均下降(P<0.01),组织损伤明显减轻。结论:YHTJF可有效减轻小鼠缺血/再灌注性肺损伤,其机制可能与其抑制caspase-12通路所致凋亡有关。
Objective: To investigate the effects of Yiqi Huoxue Tongluo Jiedu Fang (YHTJF) on lung injury induced by ischemia / reperfusion and the expression of caspase-12 in mice. Methods: Seventy male C57BL / 6J mice aged 10-12 weeks old were weighed 21-25g and were duplicated in a model of I / R injury. The rats were randomly divided into 7 groups: control group, CMC-Na group, sham group, I / R model group, low YHTJF group, L, Middle, M and H concentrations. The rats in CMC-Na, sham and I / R groups were given intraperitoneal injection of CMC-Na solution for 7 consecutive days and YHTJF-L, YHTJF-M and YHTJF-H groups were injected intraperitoneally with low, medium and high concentrations of YHTJF solution . At the end of surgery, the left lung was taken to determine the lung wet / dry ratio (W / D) and the total lung water content (TLW), and the alveolar damage assessment (IQA). The morphology and ultrastructure of the lung were observed under light and electron microscope. Tissue apoptosis index (AI) was measured by TUNEL method. The mRNA and protein expression of caspase-12 and GRP78 were detected by RT-PCR and Western blot, respectively. RESULTS: Compared with the control group, the W / D, TLW, IQA and AI were significantly increased (P <0.01) in the I / R group and the morphology of the lung tissue was significantly damaged. The mRNA and protein expressions of caspase-12 and GRP78 were increased P <0.01). However, there was no significant difference between CMC-Na group and sham group in the control group. Compared with the I / R group, the indexes of YHTJF-L, YHTJF-M and YHTJF- Outside) decreased (P <0.01), tissue injury significantly reduced. Conclusion: YHTJF can effectively reduce the ischemia / reperfusion lung injury in mice, and its mechanism may be related to the inhibition of apoptosis induced by caspase-12 pathway.