| Objective:Discuss the regulation of Choerospondias axillaris on immune function in mice,and the effect of Choerospondias axillaris on upper urinary calculi based on inflammatory immune response.Method:1.The regulation of the immune function of the Choerospondias axillaris on the immune function of mice:（1）The effect on the immune function of normal mice:randomly divide the mice into blanks,and the water extract of the Choerospondias axillaris high,medium,low（20,10,5g/kg）dose group,the drug group was given the corresponding amount of drugs.The lymphocyte suspension was prepared and cultured for 72 hours to determine the proliferation of lymphocytes.The cells were co-cultured with K562 cells for 4 hours to measure the activity of NK cells.The mice were immunized with sheep red blood cells（SRBC）,and the lymphocyte suspension antibody-forming cells were determined by spectrophotometry.The number,serum hemolysin,peripheral blood determination of peripheral blood cell classification and counting indicators.（2）Effects on immune function of immunosuppressive mice:Randomly divided mice into blank,model,high,medium low dose groups,except for the blank group,established immunosuppression by cyclophosphamide（80mg/kg）Model,drug group given the corresponding amount of drugs.Determine the corresponding immune index,the same as（1）.（3）Effects on immunological function of mice with hyperthyroidism:The mice were randomly divided into blank,model,high,medium and low dose groups,and other groups except the blank group were established with levamisole（100mg/kg）to establish an immune hyperactivity model.Model,drug group given the corresponding amount of drugs.Determine the corresponding immune index,the same as（1）.2.Effects of water extract of Choerospondias axillaris on inflammatory immune response in mice with calculus:Randomly divided mice into blank,model,high,medium,low dose,positive group（dangshi tablets）,except for blank group The other groups were given a mouse lure agent（0.75%ethylene glycol+0.75%ammonium chloride+10%glucose）to establish a model of upper urinary calculi.The drug group was given the corresponding amount of drugs,and the corresponding immune index was determined,the same as 1.1;IL-6,TNF-α,IFN-γ,IL-4 content inflammation index;kidney tissue was taken for pathological observation;Western blot was used to determine the expression of OPN in kidney tissue.3.The effect of water extract of Choerospondias axillaris on HKC crystal damage model:HKC crystal damage model was established by COM crystal intervention,and the Choerospondias axillaris water extract containing drug serum was intervened.MTT assay was used to determine cell viability.The supernatants of LDH,IL-6,TNF-α,IFN-γand IL-4 were detected by Western Blot.The expression of OPN in HKC cells was determined by Western Blot.The effect of water extract of Choerospondias axillaris on the RAW264.7inflammatory model in.4.The RAW264.7 inflammatory model was established by LPS induction,and the serum containing the Choerospondias axillaris extract was used to intervene.The cell viability was determined by MTT assay,and the IL-6 and NO content of the cell supernatant were determined by the kit.Results:1.Immunity to normal mice:（1）Compared with the blank group,the thymus and spleen organ coefficients of the three dose groups did not change significantly（P>0.05）;there was no significant change in the proliferation and transformation ability of B and T lymphocytes.（P>0.05）;the NK cell killing ability of the three dose groups was significantly decreased（P<0.05/P<0.01）,the number of antibody-forming cells was significantly increased（P<0.01）;the low-dose group was elevated with serum hemolysin（P<0.05）,the other doses were not significant（P>0.05）;there were no significant changes in peripheral blood WBC,RBC,HGB,Lym PH%,MONO%,NEUT%,EO%in the three dose groups（P>0.05）;The auricle weight difference of the sensitivity group was significantly increased（P<0.01）.Compared with the model group,the auricle weight difference of the three dose groups did not change significantly（P>0.05）.（2）Effect on immunity of immunosuppressed mice:Compared with the model,the spleen organ coefficient of the middle dose group was significantly increased（P<0.05）;the middle and low dose thymus organ coefficients were significantly increased（P<0.05）;three doses The proliferation of group B cells was significantly increased（P<0.05）;the killing power of NK cells was increased in three dose groups（P<0.05）;the number of antibody-forming cells in three dose groups was significantly increased（P<0.05）;high and medium dose groups Serum hemolysin levels were significantly increased（P<0.05）;the number of WBCs in the three dose groups was significantly increased（P<0.05）,and other indicators in peripheral blood were not significantly changed（P>0.05）.（3）The effect of water extract of five-eye fruit on immune immunity in mice:Compared with the model group,the spleen organ coefficient of the high-dose group was significantly decreased（P<0.05）;the thymus organ coefficient of the three dose groups was not significantly changed（P<0.05）;The proliferation and transformation of T lymphocytes in the middle and low dose groups were significantly decreased（P<0.05）;the killing power of NK cells in the three dose groups was not significantly changed（P>0.05）;the number of antibody formation in the three dose groups was significantly decreased.（P<0.05）,the serum hemolysin levels in the three dose groups were significantly lower（P<0.05）;the high dose groups WBC,NEUT%,EO%（P<0.05）were significantly lower,and the low dose group NEUT%was significantly increased（P<0.05）.The effect of water extract of Guangxi five-eye fruit on inflammatory immune response in mice with stone model:Compared with the model group,the content of urinary Ca2+and Cr in the high dose group was significantly decreased（P<0.05）,and the urinary acid content in the three dose groups was significantly decreased.（P<0.05）,BUN content decreased（P>0.05）;thymus organ coefficient decreased significantly in low dose group,renal organ coefficient decreased significantly in high dose group（P<0.05）,medium and low,positive group decreased The trend of serum Ig G was significantly increased in the three dose groups（P<0.01）.The serum Ig A levels in the positive and low dose groups were significantly increased（P<0.05）;the three dose groups were positive.The number of white blood cells was significantly increased（P<0.01）,and NEUT%was significantly increased（P<0.01）.In the three dose groups,the positive group Lym PH%decreased significantly（P<0.05/P<0.01）;in the three dose groups,the positive group IL-6 content decreased（P>0.05）,high,medium dose group TNF-The content ofαdecreased（P>0.05）,the positive group significantly decreased the content of TNF-α（P<0.05）,and the dose of IFN-γwas significantly decreased in the three groups（P<0.01）.The IL-4/IFN-γlevel was significantly increased（P<0.05）,and the visceral pathology score was significantly lower（P<0.05）.In the blank group,no calcium oxalate COD crystals（four-corner biconical shape）were observed.In the model group,a large amount of COD crystals appeared in the urine of the mice,high,medium and low dose groups,and the number of urine COD crystals in the positive group.Reduced,particle size is reduced.The expression of OPN protein in the kidney tissue of each group was detected by Western Blot.The expression of OPN in the kidney tissue of the blank group was weak.The expression of OPN protein in the kidney tissue of the model group was higher than that of the blank group（P<0.05）.The expression of OPN in the positive group was significantly lower than that in the three dose groups of the model group（P<0.05）.3.The effect of drug-containing serum of Choerospondias axillaris extract on HKC crystal damage model:Compared with the model group,the survival rate of middle and low dose groups increased significantly（P<0.05）.The LDH content in the supernatant of the three dose groups was significantly decreased（P<0.01）.The IL-6 content in the supernatant of the low dose group was decreased（P>0.05）.Crystal microscopic images revealed crystal particles,and no significant changes in cell morphology were observed.The effect of drug-containing serum of five-eye fruit extract on RAW264.7inflammation model:Compared with the model group,the IL-6 content in the high and middle dose groups was significantly lower（P<0.05）.High,medium and low doses significantly reduced NO content（P<0.01）.Conclusion:1.Choerospondias axillaris water extract has two-way regulation on mouse immune function:（1）It has little effect on the overall immune function of normal mice.（2）For the immunosuppressive model,increase the thymus spleen organ coefficient;enhance the B cell transformation and proliferation ability,NK cell lethality;increase the number of antibody forming cells;serum hemolysin content;increase the number of WBC,and achieve the effect of up-regulating immunity.（3）For the hyperimmune model,reduce the spleen organ coefficient;reduce the proliferation and transforming power of T lymphocytes;reduce the number of antibody formation;reduce serum hemolysin content;increase NEUT%,decrease WBC,NEUT%,EO%;The role of immunity.2.The occurrence of kidney stones in mice is accompanied by an inflammatory immune response.The water extract of the Choerospondias axillaris can increase serum Ig G and Ig A levels;increase WBC,NEUT%;decrease Lym PH%;decrease IL-6,TNF-α,IFN-γcontent;elevated IL-4/IFNγ,reduced the inflammatory immune response in model mice,inhibited the high expression of OPN,reduced the purpose of eliminating kidney stones in mice.3.Choerospondias axillaris extract containing drug serum can reduce the LDH,IL-6 content of HKC crystal damage model cell,improve cell survival rate,reduce HKC crystal damage and its induced inflammatory response,inhibit OPN high Expression,to achieve reduction,to eliminate the purpose of HKC crystal damage.4.Choerospondias axillaris water extract containing serum can reduce the inflammatory response of the inflammatory model by reducing the IL-6 and NO levels in the supernatant of RAW264.7 inflammatory model cells. |
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