摘要
目的:观察桃红四物汤对大鼠肢体缺血-再灌注损伤骨骼肌长链非编码RNA H19(LncRNA H19)、核因子κB-p65(NF-κB-p65)表达的影响。方法:选取2月龄雄性SD大鼠78只,随机分成空白组、模型组、桃红四物汤组、LncRNA H19阻滞剂组,其中空白组6只,其余每组24只。除空白组外,其他各组均阻断血流复制左后肢缺血-再灌注损伤模型,桃红四物汤组在造模前予以桃红四物汤灌胃,LncRNA H19阻滞剂组予以LncRNA H19阻断剂尾静脉注射预处理。HE染色法观察复灌后4 h腓肠肌形态及病理性改变,DAPI法观察复灌后4 h各组腓肠肌细胞凋亡变化。RT-qPCR检测复灌后(0、2、4、8 h)腓肠肌LncRNA H19、NF-κB-p65 mRNA表达,Western blot法检测0、2、4、8 h腓肠肌NF-κB-p65表达。结果:HE染色结果显示,桃红四物汤组和LncRNA H19阻断剂组肌纤维破坏及炎症浸润程度较模型组减轻。DAPI荧光染色结果显示,桃红四物汤组、LncRNA H19阻断剂组细胞凋亡率低于模型组;桃红四物汤组细胞凋亡率明显低于LncRNA H19阻断剂组,差异有统计学意义(均P<0.05)。RT-qPCR和Western blot法检测结果显示,与同一时间点空白组比较,模型组、桃红四物汤组、LncRNA H19阻断剂组LncRNA H19、NF-κB-p65表达上调,差异有统计学意义(均P<0.05)。与同一时间点模型组比较,桃红四物汤组、LncRNA H19阻断剂组、NF-κB-p65表达下调,差异有统计学意义(均P<0.05),并且不同时间点显示相同倾向性。结论:桃红四物汤可降低肢体缺血-再灌注损伤骨骼肌中LncRNA H19-NF-κB信号通路表达,减少骨骼肌细胞凋亡,减轻肢体缺血-再灌注损伤。LncRNA H19可能是大鼠肢体缺血-再灌注骨骼肌损伤病理机制中的关键上游分子。NF-κB-p65蛋白可能是大鼠肢体缺血-再灌注骨骼肌损伤病理机制中的关键蛋白。
Objective:To observe effects of Taohong Siwu decoction on the expression of long non-coding RNA H19(LncRNA H19)and nuclear factorκB-p65(NF-κB-p65)in skeletal muscle of rats with limb ischemia-reperfusion injury.Methods:78 SD rats of 2-month-old male were randomly divided into blank group,model group,Taohong Siwu decoction group and LncRNA H19 blocker group.There were 6 rats in the blank group and 24 rats in other groups.Except the blank group,the left hind limb ischemia-reperfusion injury model were made by blocking blood flow.The Taohong Siwu decoction group were given Taohong Siwu decoction intragastric administration before modeling,and LncRNA H19 blocker group given LncRNA H19 blocker tail vein injection for pretreatment.Morphological and pathological changes of gastrocnemius were observed by HE staining and apoptosis of gastrocnemius cells were observed by DAPI method at 4 h after reperfusion.The mRNA expression of LncRNA H19 and NF-κB-p65 in gastrocnemius muscle was detected by RT-qPCR(0,2,4,8 h),and the expression of NF-κB-p65 in gastrocnemius muscle were detected by Western blot(0,2,4,8 h).Results:HE staining showed that muscle fiber destruction and inflammatory infiltration were reduced in Taohong Siwu decoction group and LncRNA H19 blocker group compared with the model group.DAPI fluorescent staining results showed that the apoptosis rate of the Taohong Siwu decoction group and the LncRNA H19 blocker group were significantly lower than those in the model group,and the apoptosis rate of the Taohong Siwu decoction group was significantly lower than that of the LncRNA H19 blocker group(all P<0.05).Rt-qPCR and Western blot showed that compared with the normal group at the same time point,the expression of LncRNA H19 and NF-κB-p65 in the model group the Taohong Siwu decoction group and the LncRNA H19 blocker group were up-regulated(all P<0.05).Compared with the model group at the same time point,NF-κB-p65 expression were down-regulated in the Taohong Siwu decoction group and the LncRNA H19 blocker grou
作者
刘宗义
朱付平
李武平
LIU Zongyi;ZHU Fuping;LI Wuping(The First Hospital of Hunan University of Chinese Medicine,Changsha 410007,China)
出处
《陕西中医》
CAS
2022年第9期1163-1169,共7页
Shaanxi Journal of Traditional Chinese Medicine
基金
国家自然科学基金资助项目(81674008)
湖南省自然科学基金资助项目(2020JJ4070)
湖南省教育厅科学研究基金资助项目(20C1426)
湖南省卫生健康委员会重点课题(202204072465)。
