Exploring The Mechanism Of Limb Ischemia-reperfusion Injury And The Intervention Effect Of Taohong Siwu Decoction Based On LncRNAH19-NF-κB Signaling Axi

Objective: To observe the effect of Taohong Siwu Decoction on the expression of LncRNA H19 and NF-KB P50 in limb ischemia-reperfusion injury and the apoptosis of skeletal muscle cells,and to explore the mechanism of LncRNA H19-NF-κB signal axis in limb ischemiareperfusion injury.Methods: Seventy-eight 2-month-old male SD rats were randomly divided into control group,model group,H19 blocker group and Taohong Siwu decoction group.The control group was fed normally,and the rest groups were treated with limb ischemia-reperfusion injury model.Before modeling,the H19 blocker group was injected with H19 blocker through the tail vein for intervention;Before modeling,the peach red Siwu decoction group was given peach red Siwu decoction by intragastric administration,and the remaining groups were given the same volume of distilled water by intragastric administration.Skeletal muscle tissue of rats was taken from all animals at 0h,2h,4h and 8h at different time points after reperfusion injury.It was used to detect the expression of NF-κB P50 protein by Western-blot,the expression of H19 and NF-κB P50 mRNA by RT-PCR,the morphological and pathological changes of skeletal muscle were observed by HE staining and electron microscopy,and the apoptosis of skeletal muscle cells was observed by DAPI.Results : HE staining showed that the control group was normal.Except the control group,muscle cells in other groups were damaged to varying degrees,with the most serious injury in the model group,and the injury degree of H19 blocker group and Taohong Siwu decoction group was roughly the same.The results of electron microscopy showed that the muscles of the control group were tightly arranged without fracture,and the mitochondrial structure was clear and complete.The muscle arrangement of H19 blocker group and Taohong Siwu decoction group was slightly disordered,and the structure of the muscle was not significantly changed,while the muscle arrangement was disordered and distorted in the model group.DAPI fluorescence staining results showed that compared with control group,apoptosis rate of skeletal muscle cells in model group,H19 blocker group and Peach red Siwu decoction group was significantly higher than that in control group,the difference was statistically significant(P <0.05).Compared with model group,the apoptosis rate of skeletal muscle cells in H19 blocker group and Peach red Siwu decoction group was significantly lower than that in model group,with statistical significance(P <0.05).RT-qPCR and Western-blot assay results showed that compared with the control group at the same time point,mRNA expressions of H19 and NF-κB P50 in model group,H19 blocker group and Peach red Siwu decoction group were increased,and the expression of NF-κB P50 protein was also increased.The difference was statistically significant(P<0.05).Compared with model group at the same time point,mRNA expressions of H19 and NF-κB P50 in H19 blocker group and Taohong Siwu decoction group were decreased,and the expression of NF-κB P50 protein was also decreased,with statistical significance(P<0.05).Different time points show the same tendencyConclusion :1 LncRNA H19 is a key molecule in LI-RI.2 NF-κB P50 is a key protein in LI-RI.3 Peach red Siwu Decoction can affect the expression of NF-κB P50 and reduce the apoptosis rate of skeletal muscle cells by regulating LncRNA H19,so as to have an effect on limb ischemia-reperfusion injury...