Study On The Immunomodulatory Effect Of Hydrolyzed Ganoderma Lucidum Powder On Mice Model Of Deficiency Of Qi And Yin

Objective:To observe the changes of superoxide dismutase(SOD)activity,reduced glutathione(GSH)and trace malondialdehyde(MDA)content in lung tissues of mice with deficiency of both Qi and Yin syndrome.The contents of interleukin-2(IL-2)and interleukin-4(IL-4)in serum of model mice were observed.The mRNA expression levels of interferon-γ(IFN-γ)and Interleukin 10(IL-10)in spleen of model mice were observed.The number of CD4+T cells and the proportion of CD4+IFN-γ+and CD4+IL-4+cells to CD4+T cells in spleen lymphocytes of model mice were observed,and the possible immunomodulatory effect of hydrolyzed ganoderma lucidum powder on model mice with Qi and Yin deficiency syndrome was investigated,which provided a theoretical basis for the clinical application of hydrolyzed ganoderma lucidum powder.Methods:①Grouping and modeling of experimental animals:After adaptive feeding for 1 week,mice were randomly divided into 5 groups:normal group,model group of Qi and Yin deficiency,Shengmaisan group,hydrolyzed ganoderma lucidum powder low-dose group,hydrolyzed ganoderma lucidum powder high-dose group,with 10 mice in each group.Modified fumigation method[5][6]was used to prepare:except for the normal group,the other 4 groups were fumigated,twice a day,once in the afternoon,30min/time,for 15 days.Mold making was carried out simultaneously with drug administration.Before model making,medication was given every day,according to Shengmai Powder 2g/mL,hydrolyzed ganoderma lucidum powder at low dose of 370mg/(kg·d),hydrolyzed ganoderma lucidum powder at high dose of 1480mg/(kg·d).Once a day,0.5ml each time.Mice in normal group and model group were given the same amount of normal saline.②Model evaluation:The model of Qi and Yin deficiency induced by smoking method was evaluated from the aspects of changes in general state and signs,body weight,food intake,spleen index,bronchial histopathological changes,and "measuring the syndrome by prescription".③The SOD activity,GSH and MDA content in lung tissues of mice were detected.④The contents of IL-2 and IL-4 in serum of mice were detected by ELISA.⑤ The mRNA expression levels of IFN-γ and IL-10 in spleen lymphocytes of mice were detected by real-time qPCR.⑥ Flow cytometry was used to detect the number of CD4+T cells and the proportion of CD4+IFN-γ+and CD4+IL-4+cells to CD4+T cells in mice spleen lymphocytes.Results:①The mouse model of Qi and Yin deficiency syndrome was successfully prepared by the improved fumigation method.Through observation and analysis,hydrolyzed ganoderma lucidum powder can significantly improve the general state,body weight,food intake,spleen index,bronchial histomathology and other observation indexes of the model mice with Qi and Yin deficiency syndrome,and the effect of high dose is more significant.②After the successful modeling of Qi and Yin deficiency model mice induced by fumigation method,SOD and GSH decreased and MDA increased.After the intervention of hydrolyzed Ganoderma lucidum powder,SOD and GSH increased and MDA decreased.The results showed that hydrolyzed ganoderma lucidum powder had strong antioxidant ability and could improve the immunity of mice.③ELISA test showed that hydrolyzing Ganoderma lucidum powder could effectively regulate the secretion of serum IL-2 and IL-4,and reverse the imbalance of Th1/Th2 cells in the model mice with deficiency of both Qi and Yin,and the effect of high dose was more significant.④RT-qPCR detection:After model building,the expression level of IFN-ymRNA in spleen lymphocytes of mice with Qi and Yin deficiency was increased,the expression level of IL-10mRNA was decreased,the ratio of IFN-γ/IL-10 mRNA expression level was increased,the balance of Thl and Th2 was shifted to Th1,and the body was in a state of chronic inflammation.After treatment with hydrolyzed ganoderma lucidum powder,IFN-γmRNA was decreased in spleen lymphocytes of model mice with high levels of Qi and Yin deficiency syndrome,IL-10 mRNA expression level was up-regulated,and Thl migration state of spleen lymphocytes of model mice was reversed,so as to relieve the chronic inflammatory state of model mice.⑤Flow cytometry test:After modeling,the number of CD4+T cells,the proportion of CD4+IFN-γ+and CD4+IL-4+cells in CD4+T cells,and the ratio of CD4+IFN-γ+/CD4+IL-4+in spleen lymphocytes of the model mice were significantly increased,suggesting the imbalance of Th1/Th2 in lymphocytes of the mice.Th cells were mainly migrated towards Thl,and inflammatory factors were dominant.After treatment with hydrolyzed Ganoderma lucidum powder,the number of CD4+T cells,the proportion of CD4+IFN-γ+,CD4+IL-4+cells to CD4+T cells and the ratio of CD4+IFN-γ+/CD4+IL-4+in spleen lymphocytes of mice were significantly decreased.The results indicated that the ratio of spleen lymphocyte indexes in mice returned to the normal immune state,the immune function of mice was enhanced,and the symptoms of Qi and Yin deficiency in mice were alleviated.Conclusion:The mouse model of Qi and Yin deficiency syndrome was successfully replicated by fumigation method.Low-dose and high-dose hydrolyzed ganoderma lucidum powder could positively regulate the imbalance of Thl and Th2 in model mice,and the effect of high dose was better than that of low dose.Inflammation always runs through the whole process of pathological evolution of model mice with deficiency of both qi and Yin.The regulation effect of hydrolyzed ganoderma lucidum powder on the imbalance of Thl and Th2 in model mice is reflected by the mechanism regulation of bronchial duct inflammation in the lung of model mice.