| Purpose:The dynamic changes of immune-related factors in lung tissues,spleen tissues and serum were examined to investigate the changes of immune function in severe Mycoplasma pneumoniae pneumonia(SMPP)mice in vivo,and the TCM treatment of nourishing Yin and moistening dryness was compared with T lymphocytes CD4+%,CD8+%,CD4+/CD8+,immunoglobulin A(Ig A),immunoglobulin G(Ig G),immunoglobulin M(Ig M)and related transcription factors B Lymphocyte-inducible maturation protein-1(Blimp1),B-cell lymphoma factor 6(Bcl6),interferon regulatory factor 4(Irf4)and X-framework binding protein 1(Xbp1)expression changes were correlated to explore the target mechanism of YQDP intervention in SMPP.Material and method: One hundred and twenty BALB/c mice were randomly divided into normal group,model group,YQDP group,western medicine group and combined Chinese and Western medicine group.Except for the normal group,mice in each group were anesthetized with isoflurane and nasal drops of MP bacterial solution at a concentration of 1010 CCU/ml to construct the SMPP model,and drug intervention was started after successful establishment of the model,which was taken on days 3,7,10 and 14,respectively.The general status of mice was observed;the lung index of mice was calculated;the histopathological changes of mice lung were observed under light microscope(HE staining)and their pathological scores were calculated;the contents of T lymphocytes CD4+%,CD8+% and CD4+/CD8+ in spleen tissues were detected by flow cytometry;the contents of Blimp1,Bcl6,Irf4 and Xbp1 genes in lung tissues were determined by q PCR.Western blot method was used to detect the protein expression of Blimp1,Irf4 and Xbp1 in mouse lung tissues;ELISA method was used to detect the content of immunoglobulin Ig A,Ig G and Ig M in mouse serum.Results:1.From the observation of the general status of mice,the infection status of mice in each group was most significant on day 7 after the establishment of the model,with vertical hair,increased nasal secretion and decreased body mass,etc.On day 14,the general status of mice in the model group was improved compared with the previous one.2.Comparing the lung indices of the mice in each group,we found that the lung indices of the mice in the model group were higher than those in the normal group at all time points after MP infection,and reached the highest level on day 7(P < 0.05),and then gradually decreased.3.After the successful establishment of the model,the histopathological changes of the lung of each group were observed under light microscope and the pathological scores were compared.it was found that the alveolar structure of the mice was damaged after the infection,and the pathological scores of all groups were lower than those of the model group after the drug intervention(P < 0.05),and were highest on day 7.the pathological scores of the YQDP group were slightly lower than those of the Western medicine group at each time point,but the difference was not significant(P > 0.05).After 10 days of administration,the lung histopathological structure of each drug group showed post-injury repair,and the scores in the combined Chinese and Western medicine group were lower than those in the YQDP group except on day 3(P < 0.05),and by day 14,lower than those in the other two drug groups(P < 0.05).4.The CD4+%,CD8+% and CD4+/CD8+ contents of T lymphocytes in the spleen tissue of SMPP mice were detected by flow cytometry,and it was found that the CD4+% and CD4+/CD8+ values in the spleen of mice decreased(P < 0.05)and CD8+% increased(P < 0.05)at all time points after MP infection compared with the normal group,reaching the lowest and highest values.Compared with the model group,the CD4+% and CD4+/CD8+ values in the spleen of mice increased(P < 0.05)and CD8+% decreased to different degrees(P < 0.05)at all time points after drug administration;on day 10,the CD4+% content and CD4+/CD8+ values in the spleen of mice in the combined Chinese and Western medicine group were higher than those in the YQDP group and the Western medicine group(P < 0.05),while the CD8+% content was lower than that of the other two drug groups(P < 0.05);on day 14,the CD4+% content in the spleens of mice in the combined Chinese and Western medicine group and the YQDP group were both higher than that of the Western medicine group(P < 0.05),while the CD8+% content in the spleens of mice in the combined Chinese and Western medicine group was lower than that of the YQDP group and the Western medicine group(P < 0.05),and the CD4+/CD8+ values were higher than those of the other two drug groups(P < 0.05),but still slightly higher than the normal group(P < 0.05).5.Comparing the expression of Blimp1,Bcl6,Irf4 and Xbp1 m RNA in the lung tissues of mice in each group,we found that the relative expression of Blimp1,Irf4 and Xbp1 m RNA in the lung tissues of mice in each group after MP infection was elevated to different degrees(P < 0.05),while Bcl6 m RNA was decreased(P < 0.05).On day 7,the relative expressions of Blimp1,Irf4 and Xbp1 m RNAs in the lung tissues of mice in the combined Chinese and Western medicine group were lower than those in the other groups(P < 0.05),while the relative expressions of Bcl6 m RNA were higher than those in the Western medicine group(P < 0.05)and the Western medicine group was higher than the YQDP group(P < 0.05);on day 10,the relative expressions of Blimp1,Irf4 and Xbp1 m RNAs in the lung tissues of mice in the combined Chinese and Western medicine group were higher than those in the YQDP group(P< 0.05).On day 10,the relative expression of Blimp1,Irf4 and Xbp1 m RNA was lower than that of the other two groups(P< 0.05),and the relative expression of Bcl6 m RNA was higher than that of the other two groups(P< 0.05),and the difference between the other two groups was not significant(P > 0.05).By day 14,the relative expressions of Blimp1 and Xbp1 m RNA in the lung tissues of mice in the combined Chinese and Western medicine group were slightly higher than those in the normal group,but the difference was not significant(P < 0.05),the expression of Irf4 m RNA was lower than those in the other two groups but still higher than that in the normal group(P < 0.05),and the expression of Bcl6 m RNA was higher than those in the other two groups but still lower than that in the normal group(P < 0.05).6.After MP infection,the expression of Blimp1,Irf4,and Xbp1 proteins in the lung tissues of mice in each group was observed and found that the expression of Blimp1,Irf4,and Xbp1 proteins in the lung tissues of mice increased after infection compared with the normal group(P < 0.05),and all groups showed different degrees of decrease after drug administration(P < 0.05).By day 10 of drug administration,the relative expression of Blimp1,Irf4,and Xbp1 protein was slightly reduced in the combined Chinese and Western medicine group than in the other two drug administration groups(P < 0.05).On the 14 th day,the expression of Blimp1 and Xbp1 in the lung tissue of mice in the combined Chinese and Western medicine group was closer to that of the normal group,and the difference was not significant(P > 0.05),while the expression of Irf4 protein was still slightly higher than that of the other two groups(P < 0.05).7.Comparing the immunoglobulin contents in the serum of mice in each group,it was found that compared with the normal group,the serum Ig A,Ig G and Ig M in mice increased at all time points after MP infection(P < 0.05)and reached the highest values on day 3,day 10 and day 3,respectively;compared with the model group,the serum Ig A,Ig G and Ig M contents in mice decreased at all time points after drug administration(P < 0.05);by On day 7,the serum Ig A and Ig M contents of mice in the YQDP and combined Chinese and Western medicine groups were lower than those in the Western medicine group(P < 0.05);on day 10,the serum Ig G contents of mice in the YQDP and combined Chinese and Western medicine groups were lower than those in the Western medicine group(P < 0.05);after 14 days of administration,the serum Ig A,Ig G and Ig M contents of mice in each group were lower than those in the combined Chinese and Western medicine group(P < 0.05),but still slightly higher than the normal group.Conclusion:1.After the establishment of the SMPP model,the mice showed decreased T-lymphocyte CD4+%,CD4+/CD8+,increased CD8+% and increased immunoglobulin Ig A,Ig G and Ig M,indicating the existence of cellular and humoral immune disorders in SMPP mice,which provided a research basis to further explore the targets of TCM in regulating immune function in SMPP mice.2.YQDP can improve the in vivo immune disorder in SMPP mice by increasing the expression of T lymphocytes CD4+%,CD4+/CD8+,decreasing CD8+% and immunoglobulins Ig A,Ig G and Ig M,and also promote the expression of Blimp1,Xbp1,Irf4 and inhibit the production of Bcl6 to regulate the local immune environment,achieving the effect of nourishing Yin and moistening dryness to improve immune function. |
PDF Full Text Request