Pharmacodynamics And Mechanism Study Of Yinma Jiedu Granule In The Treatment Of Acute Lung Injury And Acute Pharyngitis

Part 1 Mechanism study of Yinma Jiedu Granule alleviating acute lung injury in rats via suppressing inflammationAcute lung injury(ALI)and its more severe form,acute respiratory distress syndrome(ARDS),are common critical diseases,with clinical manifestations of edema with alveolar and diffuse pulmonary interstitial as well as acute hypoxic respiratory insufficiency.Although the pathophysiology of ALI/ARDS has been extensively investigated,its mortality rate remains high,at approximately 40%,which seriously threatens the health and life of patients.Currently,there is no specific therapy for ALI/ARDS,and its treatment is mainly supportive therapy,so effective treatment and management of ALI/ARDS is needed urgently.In recent years,many natural products and Chinese patent medicine(CPM)have been reported to have multiple anti-inflammatory activities and lung protection effects.Yinma Jiedu Granule(YMJD)is a traditional CPM,with five Chinese herbal extracts,including Lonicerae Confusa DC.(Shanyinhua in Chinese),Portulaca oleracea L.(Machixian in Chinese),Plantago asiatica L.(Cheqiancao in Chinese),Rheum officinale Baill.(Dahuang in Chinese),Glycyrrhiza uralensis Fisch.(Gancao in Chinese),which has been proven to have anti-inflammatory effects and therapeutic effects on chronic obstructive pulmonary disease(COPD).The main purpose of the current investigation is to explore whether YMJD can alleviate ALI induced by lipopolysaccharide(LPS)in rats and its underlying mechanisms.Rats were treated orally with YMJD for 14 consecutive days,and 2 h after the last administration,the rat model of ALI was induced by the intratracheal instillation of LPS.The results showed that the clinical symptoms of ALI rats were significantly improved after YMJD treatment,manifested as increased activity and weight gain.YMJD also effectively attenuated pulmonary edema in ALI rats.HE staining showed that YMJD can significantly reduce the pathological damage of rat lung tissue and alleviate the lung inflammatory response.YMJD attenuated pulmonary vacuolization and brush border remodeling of bronchial epithelial cells as detected by transmission electron microscopy.Moreover,we found that YMJD effectively reduced the serum lactate(LA)levels in ALI rats,decreased the infiltration of white blood cells(WBC),lymphocytes(LYM),mononuclear cells(MON)and neutrophils(NEUT)in bronchoalveolar lavage fluid(BALF),reduced the concentration of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)and inhibited inducible nitric oxide synthase(iNOS)and cyclooxygenase-2(COX-2)expression in the lung tissue.Additionally,our study demonstrated that YMJD could significantly increase the activity of superoxide dismutase(SOD)and reduce the malondialdehyde(MDA)level in the lung tissue,suggesting that YMJD was capable of inhibiting inflammation and antioxidant stress in the lung tissue.By employing RNA-sequencing,we identified that JAK2/STAT1 was an important signaling pathway that involved in the lung protection of YMJD,and further Western blot assay verified that YMJD could effectively inhibit the activation of the JAK2/STAT1 inflammatory pathway in ALI rats and LPS-stimulated RAW264.7 cells.The above results indicated that YMJD treatment could improve the general state of ALI rats,reduce pulmonary edema,pulmonary inflammatory response and lung histopathological damage,highlighting that YMJD has a significant protective effect on the lung injury of ALI rats.Further mechanistic study showed that YMJD reduced the inflammatory response and oxidative stress by inhibiting the activation of the JAK2/STAT1 signaling pathway,thereby exerting a protective effect on the lung.These findings provided evidence for the clinical use of YMJD in the treatment of inflammatory pulmonary diseases such as ALI.Part 2 Pharmacodynamics and mechanism study of Yinma Jiedu Granule in alleviating acute pharyngitis in ratsAcute pharyngitis is inflammation that occurs in the pharynx,tonsils and surrounding tissues,and is one of the most common diseases in otolaryngology.Acute pharyngitis is usually a self-limiting disease that can recover within 1-2 weeks.However,acute pharyngitis caused by streptococcal infection is one of the most serious diseases in clinical practice due to its high incidence and mortality rate,as well as the potential for severe complications.The treatment of acute pharyngitis includes supportive treatment and antimicrobial therapy,the purpose of which is to alleviate pharyngeal symptoms,decrease the duration of infection,and prevent the occurrence of various complications.However,the use of antibiotics is limited due to their resistance and adverse reactions,making it necessary to develop alternative drugs for the treatment of acute pharyngitis.Traditional Chinese Medicine(TCM),as one of the main complementary and alternative medicines,has accumulated extensive experience in treating inflammatory diseases and many Chinese herbal medicines have been reported to have the effect of alleviating acute pharyngitis.Yinma Jiedu Granule(YMJD)is a Chinese patent medicine produced by the Yangtze River Pharmaceutical Group Sichuan Hairong Pharmaceutical Co.,Ltd.It has been proven to have obvious anti-inflammatory effects and clinical studies have reported that it has a significant effect on cough evil committed lung syndrome.The present study was designed to evaluate the pharmacodynamics of YMJD in the treatment of acute pharyngitis and explore its possible molecular mechanism by a rat model of acute pharyngitis induced by ammonia.Rat model of acute pharyngitis was developed by spraying 15%ammonia solution(V/V)onto the pharynx of rats.According to the severity of the pharyngeal symptoms,rats were divided into control group,ammonia-induced acute pharyngitis group,YMJD 500,1000 and 2000 mg/kg-treated group.After treating orally with YMJD for 5 consecutive days,samples were collected for testing.The results showed that YMJD could significantly alleviate the symptoms of acute pharyngitis in rats,including coughing,saliva secretion,oral hair loss,reduced activity,swelling and redness of the pharynx,etc.HE staining showed that the mucosal layer of the pharyngeal tissue became thinner,and the tissue hyperplasia and inflammatory cell infiltration decreased after YMJD administration,suggesting that YMJD could alleviate the pathological damage of the pharynx in rats.The detection results of inflammation-related indicators showed that YMJD treatment could effectively reduce the amount of WBC,LYM,MON,and NEUT in the blood of rats,and inhibit the production of serum TNF-α and LA,as well as decreased the mRNA expression of TNF-α and iNOS in pharyngeal tissue.Moreover,we also found that YMJD could significantly reduce the level of oxidative stress in the pharyngeal tissue of rats,manifested by increasing the activity of SOD and reducing the content of MDA,indicating that YMJD could inhibit inflammation and oxidative stress in rats.Further mechanistic studies have shown that YMJD could significantly reduce the phosphorylation of phosphatidylinositol 3-kinase(PI3K),AKT(also known as protein kinase B,PKB)and mammalian target of rapamycin(mTOR)in pharyngeal tissue,suggesting that YMJD might alleviate inflammation and oxidative stress by suppressing the PI3K/AKT/mTOR signaling pathway,thereby exerting protective effects on rats with acute pharyngitis.The above results demonstrated that YMJD has the potential to be developed as a new drug for the treatment of acute pharyngitis.