Study On The Acute Pharyngitis Therapeutic Effect And Mechanism Of Hydrogen

H₂ is a diatomic gas molecule with flammability and certain reducibility.In recent years,many studies have shown that H₂ has anti-oxidation,anti-inflammatory and antiapoptotic effects.It has been reported that H₂ can regulate the abnormal secretion of inflammatory factors in macrophages induced by LPS,suggesting that its preventive and therapeutic effects on many diseases may be related to its anti-inflammatory effects.However,the current research on the molecular mechanism and target of H₂’s antiinflammatory effect is not clear.Based on this,this subject will study the potential mechanism of H₂’s anti-inflammatory effect and demonstrate its therapeutic effect on acute pharyngitis through animal experiments and provide a scientific basis for the clinical promotion of H₂.In this project,the molecular target of H₂’s anti-inflammatory effect is firstly determined through cell experiments.The experimental results show that in the LPSinduced RAW264.7 cell inflammation model,H₂（at a concentration of 1.2 ppm）can regulate inflammatory factors（NO,TNF-α,IL-6,IL-10）related mRNA expression,thereby inhibiting the abnormal secretion of inflammatory factors.Further RNA sequencing analysis found that H₂ can inhibit the activation of NF-κB and MAPK signaling pathways and exert anti-inflammatory effects.Specifically,in the verification-related signal transduction pathway that LPS stimulates the activation of TLR4 receptors on the surface of macrophages,H₂ can inhibit the activation of IκBαand NF-κB,while in the MAPK pathway,H₂ can inhibit the cascade reaction and phosphorylation level of key proteins.In addition,in the NF-κB and MAPK pathways,we have newly discovered that H₂ also acts on the signaling molecule PKC8 and the heterodimer AP-1,which has not been clearly reported in previous H₂ medical studies.In order to further confirm the feedback information of RNA sequencing analysis,we use Western Blot to quantitatively analyze the signal target.The results show that H₂ can inhibit phosphorylation of IκBα and the translocation of NF-κB p65 from the cytoplasm to the nucleus,thereby inhibiting the activation of the NF-κB pathway;while in the MAPK signaling pathway,H₂ can significantly inhibit the phosphorylation level of P38,ERK1/2,JNK,and finally inhibit the activation of the heterodimer AP-1 composed of c-Jun and c-Fos.After clarifying the mechanism of H₂ at the cellular level,we used animal experiments to demonstrate the anti-inflammatory effect of H₂ in vivo.The rat pharynx was stimulated with ammonia water to construct an acute pharyngitis model,and then 2%and 4%H₂ treatment were given respectively.The results showed that the stimulation of ammonia water caused the rats to show the symptoms and signs of acute pharyngitis,and both 2%and 4%H₂ improved the symptoms of the rats.H₂ can reduce rat pharyngeal mucosal thickening,inflammatory cell infiltration and glandular hypertrophy,and can significantly regulate the abnormal secretion of inflammatory factors PGE2,TNF-α,IL-6,IL-10 in the serum.It has a significant inhibitory effect on the transcription level of NF-κB and MAPK signaling pathways and shows good anti-inflammatory activity in vivo.In summary,the mechanism by which H₂ exerts an anti-inflammatory effect is to inhibit the activation of NF-κB and MAPK signals,and then regulate the transcription and translation of downstream related inflammatory factors,thereby alleviating the clinical symptoms caused by inflammation.This research is based on previous studies and demonstrates the reliability of previous H₂ medical research results through cell experiments.At the same time,it also puts forward our latest research viewpoints and molecular targets for its role,thereby further improving the H₂ anti-inflammatory molecule way.NF-κB and MAPK are two classic signaling pathways.These two signaling pathways play an important role in biological processes such as inflammation,oxidation,and apoptosis.H₂ has strong diffusivity and acts on these two signaling pathways.It also suggests that H₂,as a new type of medical gas molecule,has potential application value in other related diseases involving NF-κB and MAPK signaling pathways.