| Object i ve1.To construct a BALB/c nude mouse model infected with SIVmac251.2.To observe the changes of SIV viral load in peripheral blood of the model mice.3.Observe the pathological changes of the main organs of the model mice.4.To explore the effect of highly active anti retroviral therapy(HAART)and Bushen Jiedu compound on the viral load in peripheral blood of mice.5.To observe the effects of Bushen Jiedu compound and HAART on the body weight,spleen mass and spleen index of the model mice.6.To observe the effect of Bushen Jiedu compound and HAART on peripheral blood lymphocyte and IgG antibody of the model mice.Methods1.Using cell culture technology,the virus was infected with tzm-bl cell line,and the suspension of tzm-bl cells infected with SIV was injected into BALB/C nude mice by intraperitoneal injection.2.The detection of SIV virus in peripheral blood and the change of SIV viral load were carried out by using mandible blood sampling method and reverse transcription real-time fluorescence quantitative PCR(RT-PCR).3.Paraffin section technique and hematoxylin eosin staining(he)were used to observe the histopathological changes of mice.4.L part of Y,y part of Y,s part of G and s part of L were extracted by phytochemical organic solvent extraction method,and the four effective parts were used to form the Bushen Jiedu compound.5.Flow cytometry(FCM)and blood routine were used to detect the proportion and count of peripheral blood lymphocytes.6.IgG antibody in peripheral blood was detected by enzyme linked immunosorbent assay(ELISA).Results1.SIV was detected in the peripheral blood of BALB/C nude mice on the seventh day after inoculation with the suspension of tzm-bl cells infected with SIV.The detection rate was 100%.SIVmac251 viral load was detected for seven consecutive weeks.The viral load in plasma continued to rise in the first five weeks,reached the maximum value in the fifth week,and then decreased gradually.2.At the seventh week after modeling,the pathological changes of main organs in BALB/C nude mice were observed.The results showed that there were inflammatory cell infiltration in portal area of liver tissue,some normal lung tissue structure disappeared,lung tissue necrosis,some alveolar filled with serous,red blood cells and a small amount of neutrophils,some brain tissue necrosis,brain tissue structure became loose,tissue edema,model group In nude mice,there were a few structural changes in spleen.3.After continuous administration of HAART for one week,there was no significant difference in plasma viral load between the low-dose HAART group and the model group(P＞0.05),but there was significant difference between the high-dose HAART group and the model group(P<0.05).After two weeks of continuous administration,there was no significant difference between the low-dose group and the model group(P＞0.05),but there was significant difference between the high-dose group and the model group(P<0.05).4.One week and two weeks after stopping gavage,the viral load in plasma of HAART low-dose group and high-dose group was significantly different from that of model group(all P ＜0.01),which indicated that HAART could still reduce the viral load of the model within two weeks.5.After continuous gavage of Bushen Jiedu compound for one week,the total number of peripheral blood lymphocytes in the high-dose Bushen Jiedu compound group was significantly higher than that in the model group and blank group(P ＜0.05).After two weeks of continuous administration,the low-dose Bushen Jiedu compound group was significantly different from the model group and blank group(all P<0.01),indicating that the Bushen Jiedu compound can improve the total number of peripheral blood lymphocytes in model mice and blank mice.6.Compared with model group and blank group,the ratio of T lymphocyte in peripheral blood of high dose group was significantly higher than that of model group and blank group(all P ＜0.01);the ratio of T lymphocyte in low dose group was not significantly different from that in model group and blank group Compared with model group and blank group,HAART western medicine group could significantly increase the proportion of peripheral blood T lymphocytes(P ＜0.01).Compared with the model group,the high-dose group,the low-dose group and the model group of Bushen Jiedu compound could significantly increase the proportion of B-lymphocyte in the peripheral blood of the model mice,and the differences were statistically significant(P<0.01).Compared with the high-dose group,low-dose group and blank group,the B-lymphocyte ratio in peripheral blood of BALB/C nude mice was significantly increased Compared with model group,HAART western medicine group could improve peripheral blood B lymphocyte,the difference was statistically significant(P ＜0.05).The results showed that Bushen Jiedu compound could improve the ratio of T and B lymphocytes in peripheral blood of model mice and blank mice,and the effect of Chinese medicine was better.7.After one week of continuous gavage with Bushen Jiedu compound,the total number of B lymphocytes in peripheral blood of high-dose Bushen Jiedu compound group was significantly higher than that of blank group(P ＜0.01).Compared with blank group,the difference was statistically significant(P＜0.05).Two weeks after intragastric administration,the total number of B lymphocytes in the low-dose Bushen Jiedu compound group was significantly higher than that in the model group and the blank group(P<0.05),and the difference was statistically significant.It showed that the Bushen Jiedu compound could improve the total number of B lymphocytes in peripheral blood of model mice and blank mice.After continuous intragastric administration for one week,the total number of NK cells in the high-dose Bushen Jiedu compound group was significantly higher than that in the model group and blank group(P ＜0.05).After a week of administration,the total number of NK cells in the low-dose Bushen Jiedu compound group was significantly higher than that in the model group and blank group(P ＜0.05),indicating that the Bushen Jiedu compound can improve the total number of NK cells in peripheral blood of model mice and blank mice.8.After two weeks of intragastric administration,the levels of IgG in peripheral blood of high-dose group and low-dose group of Bushen Jiedu compound were compared with those of model group(P<0.05).One week after drug withdrawal,there were statistically significant differences between low-dose Bushen Jiedu compound group,HAART western medicine group and model group(P＜0.05).The results showed that Bushen Jiedu compound and HAART western medicine could increase the IgG concentration in peripheral blood of model mice,and the effect of Bushen Jiedu compound was better than HAART western medicine.9.Two weeks later,the body weight of mice in high-dose Bushen Jiedu compound group and HAART western medicine group was lower than that of model group(P ＜0.05);spleen mass of mice in high-dose and low-dose groups of Bushen Jiedu compound,HAART western medicine group and blank group was significantly lower than that of model group(P ＜0.01)The spleen index of mice in dose group,low-dose group and HAART western medicine group was significantly lower than that of model group(P ＜0.01);spleen index of model group was significantly higher than that of model group(P ＜0.01).The results showed that the spleen weight and spleen index of model group were higher than those of blank group,while Bushen Jiedu compound and HAART western medicine could inhibit splenomegaly and decrease spleen index.Conclusion1.After one week,SIV was detected in the peripheral blood of BALB/C nude mice,and SIV virus was detected in the peripheral blood of BALB/C nude mice for seven consecutive weeks.This method based on BALB/C nude mice to construct SIV infection animal model is successful,which has the advantages of repeatability,sustainable observation and simple operation.2.Through the efficacy test of highly effective combined antiretroviral therapy(HAART)composed of zidovudine(AZT),lamivudine(3TC)and EFA virenz(EFV)in this animal model,it is verified that the model can evaluate the effect of HAART on the inhibition of SIV viral load in mice.3.The kidney tonifying and detoxifying compound composed of L part,y part,s part of G and s part of 1 can increase the total number of lymphocytes in peripheral blood of model mice and blank mice,increase the ratio of T lymphocyte to B lymphocyte,increase the number of B lymphocyte and NK cell,increase the antibody concentration of IgG in peripheral blood,inhibit splenomegaly and decrease the spleen finger of model mice The role of numbers.Two weeks after stopping the drug,it showed the effect of inhibiting the viral load,which fully reflected the function of Bushen Jiedu compound in improving the immune function of mice and inhibiting the viral load.4.HAART could not increase the total number of lymphocytes in peripheral blood of model and blank mice,but could increase the ratio of T lymphocyte and B lymphocyte and the concentration of IgG antibody in peripheral blood,but its effect was not as good as that of Bushen jiedu compound.At the same time,it can inhibit splenomegaly and decrease spleen index.After two weeks of administration,it can increase the number of B lymphocytes,but the effect is not as good as the low dose of Bushen Jiedu compound,which fully shows that HAART western medicine has a certain effect on improving immunity. |
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