Ameliorative Potential Of Ginseng And/or Platycodon Grandiflorum On Inflammatory Reaction Of Lung Injury Induced By Haze Exposure

AIM:In recent years,China has repeatedly happen a large area of haze events,causing widespread concern in society.Haze not only for human daily life and travel inconvenience,but also a great threat to human health.Ginseng and Platycodon grandiflorum is often mentioned in the prescription can relieve lung injury medicine.In this study,the model of cigarette smoke was instead of haze model to explore the effect of Ginseng and Platycodon grandiflorum on lung injury and the specific regulation mechanism.METHODS:Ginseng,Radix ginseng rubra,platycodon grandiflorum were extracted by water extraction and alcohol extraction.The contents of ginsenoside and platycodin in different extracts were determined by high performance liquid chromatography(HPLC).Hepatic stellate cells(HSC)were cultured and the toxicity of the extracts was determined by MTT assay.The mouse model was established by cigarette smoke exposure method.The mice were placed in a closed plastic box(50 ×40 × 30cm),and 5 cigarettes were lit into the box.The cigarette was burned in 15 minutes.Smoke completely dispersed,while the mice rest for 5 minutes.Then reignite 5 cigarettes repeat the above steps.Smoked 3 times a day,each time smoked 1 hour/15 cigarettes and continued a total of 11 weeks.Mice received Panax Ginseng(PG),Radix ginseng rubra(RGR),platycodon grandiflorum(PG)and Ginseng+ platycodon grandiflorum combined extract by oral gavage in a dose of 300mg/kg/day for 11 weeks after starting CS exposed and until the mice were sacrificed.Mice were killed and collected blood.The liver,lung,cardiac,kidney were dissected,weighed,take pictures and snap frozen in liquid nitrogen and store at-80 ?until analyzed.A small portion of each lung was fixed with 10%formalin and embedded in paraffin.Blood were separated by centrifugation,and the serum samples were subjected to biochemical investigations.Lung sample from each mice were removed for histopathological and biochemical examination.Serum levels of hydroxyproline(HYP)and interleukin(IL)were observed.The pathological changes of lung tissue were observed by H&E,Masson and immunohistochemistry.The(a-SMA),matrix metalloproteinase-13(MMP-13),tissue inhibitor of metalloproteinase(TIMP-1)and purine can be detected by Western blot.(NF-?B),nuclear factor kappa B inhibitory protein a(I?B-?),cysteine-containing aspartic proteolytic enzyme-1(Caspase-1),cytosine(NLRP3),apoptosis-related point-like protein(ASC),Toll-like receptor 4(TLR4),acetylated enzyme-1(SIRT-1),interleukin-1 receptor-related kinase(TNF-a)and glyceraldehyde-3-phosphate dehydrogenase(GAPDH)were detected by reverse transcription polymerase chain reaction(RT-PCR),and the expression of P2X7r,IL-1?,IL-18,IL-6,TNF-a and GAPDH.RESULTS:The contents of ginsenoside and platycodin were determined by high performance liquid chromatography(HPLC).The results showed that ginseng,red ginseng,platycodon grandiflorum and ginseng + herbal extract contained a certain amount of ginsenoside and platycodin.The MTT assay of t-HSC cells demonstrated that the different extracts were not cytotoxic.Cigarette smoke exposure can significantly reduce the weight of mice,H&E,Masson results can be seen in cigarette smoke exposure to mice lung function led to mouse lung tissue inflammatory cell infiltration,lung tissue changes and emphysema occurred,and can be significantly increased The levels of IL-la,IL-1? and HYP in serum of mice.Western blot and PCR results can be seen that different extracts can significantly reduce the expression of ?-SMA,?P2x7R?NF-?B?Cas-1?NALP3?ASC?TLR4?IRAK-4?IL-1??IL-1??IL-18?IL-6?TNF-?,significantly increased I?B-?,SIRT-1 and other levels,adjust the ratio of MMP-13/TIMP-1.Among them,Campanulaceae extract and garden ginseng + Campanulaceae extract the most obvious effect.CONCLUSION:The combination of ginseng,red ginseng,platycodon grandiflorum and garden ginseng + Platycodon grandiflorum showed a better effect on the pulmonary fibrosis induced by smoke exposure.The expression of P2x7R in lung tissue of smoke exposure mice was higher than that in P2x7R,in the lungs of inflammation and damage to the core target;Ginseng Campanulaceae extract substances can be regulated by P2x7R pathway to improve cigarette smoke exposure caused by lung injury.