Study On Alleviation Of Allergic Reaction Of Mice With Urticaria-like Lesions By Danggui Yinzi Based On Autophagy Pathway Mechanism

Objective:To investigate the effect of Danggui Yinzi on allergic reaction in urticaria-like lesions mice model and the mechanism of autophagy intervention.Method:SPF-class BALB/c mice were used to replicate the urticaria-like lesions mice model by intraperitoneal injection of ovalbumin and aluminum hydroxide suspension.The animals were randomly allocated into six groups: a normal group(normal saline 4ml/kg/d),a model group(normal saline 4ml/kg/d),a loratadine group(1.3 mg/kg/d),a Danggui Yinzi high dose group(39.3g/kg/d),a Danggui Yinzi medium dose group(19.6g/kg/d)and a Danggui Yinzi low dose group(9.8g/kg/d).The pathological changes of skin tissues were observed by hematoxylin-eosin(HE)staining.Morphological changes of autophagy in skin tissues epithelial cells were observed by transmission electron microscope.The m RNA levels of the autophagy-related gene 5(ATG5),the microtubule-associated protein 1 light chain 3B(LC3B)and the SQSTM1/p62(p62)m RNA in skin tissues were detected by real-time quantitative polymerase chain reaction(RT-PCR).The expressions of ATG5,LC3 B and p62 protein in skin tissues were detected by immunohistochemistry(IHC).Results:Compared with the normal group,the skin tissue of the model group showed obvious urticaria-like pathological changes,the ultrastructure of the epithelial cells was destroyed,and a certain degree of autophagosomes appeared in the cytoplasm.Danggui Yinzi can significantly improve the pathological manifestations of dermal edema,collagen bundles separation,telangiectasia in urticaria-like lesions mice;it can also improve autophagosomes formation and abnormal cell ultrastructure such as nuclear chromatin condensation,mitochondrial swelling,endoplasmic reticulum expansion,etc.Compared with the normal group,the ATG5 protein expressions of the model group was increased;the LC3 B protein expressions was significantly increased(P<0.01);LC3B m RNA level was increased too,while p62 m RNA levels and its protein expressions were obviously decreased-regulated(P <0.05 or P <0.01).Compared with the model group,levels of ATG5 and LC3 B m RNA with their protein expressions of the Danggui Yinzi groups were significantly increased(P<0.01 or P<0.05),while p62 m RNA levels and its protein expressions were significantly decreased-regulated(P<0.05 or P<0.01).Conclusion:Danggui Yinzi can up-regulate the m RNA and protein expression of ATG5 and LC3 B,down-regulate p62 m RNA and protein expression,activates the autophagy mechanism of the skin tissue,and enhances the autophagy level to treat the pathological state of skin tissue in urticaria-like mice.