| Objective: To investigate whether the traditional Chinese medicine prescription Taohongsiwu decoction alleviates the inflammatory response by inhibiting the expression of inositol 1,4,5-trisphosphate receptor(IP3R)during ischemia reperfusion injury(Limb Ischemia Reperfusion Injury,LIRI)in rat skeletal muscle based on Wnt/Ca2 signaling pathway.Methods:40 male SD rats were randomly divided into normal control group N(normal),model group M(model)10,taohong sihuye group T(taohong siwuye)10 and blocker group B(block)10,respectively.Each group was divided into 4 groups at 2nd,4th,6th and 8 hours after ischemia reperfusion.lisa assay for the detection of inflammatory factor expression,real-time quantitative pcr and western-blotting for the detection of ip3r expression.Results:1.The morphology of gastrocnemius muscle cells was observed by HE staining: the normal control group showed long cord shape,the muscle fibers were arranged neatly,the structure was clear and the muscle marrow was abundant.several nuclei in each muscle fiber tightly attached to the inner surface of the muscle membrane.The size of the nucleus was uniform,the coloring was uniform,and there was no abnormal change;the muscle fiber in the model group of ischemia-reperfusion group was disordered,thickened,edema,flake.muscle fiber integrity,continuous destruction,thinning of muscle fibers,expansion of muscle fiber interfracture,muscle cell destruction and muscle fiber inflammatory infiltration;the skeletal muscle of Taohong Siwu Tang group was still inflammatory infiltration,but it was lighter than that of the model group.cells were damaged,arranged neatly,and inflammatory infiltration was not obvious.2.DAPI fluorescence staining method was used to detect the nucleus of gastrocnemius: the apoptotic cells in the model group were significantly higher than those in the normal group,and the number of apoptotic cells in the group was reduced.compared with the model group,the number of apoptotic cells and the fluorescence intensity in the blocker group were significantly lower than those in the model group.3.ELISA method was used to detect the expression of IL-1\946;,IL-6and TNF-? : the macrophages secreted by the inflammatory factors IL-1\946;,IL-6 and TNF-? in the model group were significantly stronger than those in the normal group.Compared with the model group,the expression of the inflammatory factors IL-1\946;,IL-6 and TNF-? in the pretreatment group of Taohong Siwu decoction was significantly lower than that in the blocker group.However,there was no significant difference in IL-6 concentration in Taohong Siwu decoction group and blocker group.4.Real-time quantitative PCR and western-blotting detection of IP3R:IP3R protein were significantly up-regulated to normal levels,and the expression of IP3R was significantly down-regulated or even reduced to normal levels after pretreatment of Taohong Siwu Tang.Conclusion: In the Wnt/Ca2 signaling pathway of skeletal muscle ischemia reperfusion injury,Taohong Siwu decoction can alleviate the inflammatory response by inhibiting the expression of IP3R,thus reducing the limb ischemia reperfusion injury. |
PDF Full Text Request