桃红四物汤通过LncRNA H19对肢体缺血-再灌注损伤中细胞凋亡的影响研究

Effect of Taohong Siwu Decoction on Apoptosis of LIRI through LncRNA H19

目的:研究桃红四物汤通过LncRNA H19对肢体缺血-再灌注损伤中细胞凋亡的影响,探讨治疗肢体缺血再灌注损伤的新思路。方法:将78只2月龄雄性SD大鼠随机分为对照组、模型组、H19阻断剂组、桃红四物汤组;对照组正常饲养,剩余各组均造成肢体缺血-再灌注损伤模型;在造模前,H19阻断剂组予以H19阻断剂,通过尾静脉注射进行干预;在造模前,桃红四物汤组予以桃红四物汤进行灌胃处理,余下各组使用相同体积的蒸馏水灌胃处理;所有动物在再灌注损伤后的0、2、4、8 h 4个不同时间点,取大鼠骨骼肌肌肉组织,RT-PCR法检测骨骼肌中H19的mRNA的表达,HE染色法和电镜技术观察骨骼肌形态及病理性改变,DAPI法观察骨骼肌细胞凋亡情况。结果:在相同的时间点与对照组相比,模型组、H19阻断剂组、桃红四物汤组的H19 mRNA水平均升高,差异有统计学意义(P<0.05);在相同的时间与模型组相比,H19阻断剂组、桃红四物汤组的H19 mRNA水平均降低,差异有统计学意义(P<0.05);与对照组相比,模型组、H19阻断剂组和桃红四物汤组骨骼肌细胞凋亡率明显高于对照组,差异有统计学意义(P<0.05);与模型组相比,H19阻断剂组和桃红四物汤组骨骼肌细胞凋亡率明显低于模型组,差异有统计学意义(P<0.05)。结论:LncRNA H19是肢体缺血再灌注损伤中的重要分子,抑制LncRNA H19的表达可以降低骨骼肌细胞的凋亡率,从而治疗肢体缺血再灌注损伤。

Objective:To study the effect of Taohong Siwu Decoction on apoptosis of limb ischemia-reperfusion injury(LIRI)through LncRNA H19,and to explore a new idea for the treatment of LIRI.Methods:78 SD rats of 2-month-old were randomly divided into the control group,the model group,the H19 blocker group and the Taohong Siwu Decoction group.Before modeling,the H19 blocker group was intervened with injection of H19 blocker through the tail vein,and the Taohong Siwu Decoction group was intervened with Taohong Siwu Decoction.The expression of H19 mRNA in skeletal muscle was detected by RT-PCR at 0 h,2 h,4 h and 8 h after reperfusion injury.The morphological and pathological changes of skeletal muscle were observed by HE staining and electron microscopy.The apoptosis of skeletal muscle cells was observed by DAPI.Results:Compared with that in the control group at the same time point,the mRNA level of H19 was increased in the model group,the H19 blocker group and the Taohong Siwu Decoction group(P<0.05);compared with that in the model group,the mRNA level of H19 was decreased in the H19 blocker group and the Taohong Siwu Decoction group(P<0.05).Compared with that in the control group,the apoptosis rate of skeletal muscle cells was significantly increased in the model group,the H19 blocker group and the Taohong Siwu Decoction group(P<0.05);compared with that in the model group,the apoptosis rate of skeletal muscle cells was significantly decreased in the H19 blocker group and the Taohong Siwu Decoction group(P<0.05).Conclusion:LncRNA H19 is an important molecule in LIRI,Siwu Decoction can inhibit LncRNA H19 expression to reduce apoptosis rate of skeletal muscle cells,so as to treat LIRI.