| Danggui Buxue Tang is a prescription for nourishing Qi and nourishing blood created by Li Dongyuan during the Jin and Yuan Dynasties.It is composed of Astragalus and Angelica two herbs in a ratio of 5:1.It has the effect of nourishing Qi and nourishing blood.The heat syndrome of Yang.A large number of studies have confirmed that oral Danggui Buxue Tang decoction can enhance the non-specific immune response in the body’s pathological state,but the immune enhancement effect of the vaccine has not been reported,so this study immunized normal mice with foot-and-mouth disease(FMD)vaccine to explore oral Danggui Buxue Tang The effect on the immune effect of FMD vaccine and the immune function of intestinal mucosa provides a theoretical reference for further development as an oral immune adjuvant.The main tests are as follows:1.Immunoenhancing effect of oral Danggui Buxue Tang on O-type FMD vaccine.In order to evaluate the immunomodulatory effect of oral Danggui Buxue Tang on FMD vaccine,70Kunming mice were random Ly divided into 7 groups of 10 mice,namely Danggui Buxue Tang 2 g/m L,1g/m L,0.5 g/m L,0.25 g/m L,0.125 g/m L,0.0625 g/m L dose group and control group(saline);the intragastric dose was 0.25 m L/d for 5 days.Twenty-four hours after the last administration,all test mice were injected subcutaneously with 0.2 m L type-O of FMD vaccine in the groin.Separate blood was collected from the submandibular blood 1 and 3 week after the second immunizations,and the levels of serum specific Ig G and antibody subclasses Ig G1,Ig G2a,Ig G2b and Ig G3 at 1 and 3 week after the second immunizations in each test mouse were determined by ELISA;Mice were sacrificed by neck removal at the 3rd week after the second immunization,and the spleen and thymus were harvested to measure the organ index.The results showed that compared with the control group,the body weight and thymus and spleen indexes of the test mice did not change significantly after oral administration of Danggui Buxue Tang,and the difference was not statistically significant(P>0.05);Serum total Ig G content increased significantly in the 1 week after the second immunization(P<0.01);serum total antibody Ig G and antibody subclass Ig G1 in 0.5 g/m L and 2g/m L dose groups in the third week after the second immunization The content increased significantly(P<0.01),the content of antibody subclass Ig G2a in the 0.5 g/m L dose group was significantly increased(P<0.01),and the content of Ig G2b and Ig G3 had no significant changes compared with the control group(P>0.05).It is suggested that oral Danggui Buxue Tang can enhance the immune effect of O-type FMD vaccine,and 0.5 g/m L dose is the best.2.Effect of oral administration of Danggui Buxue Tang on the m RNA expression of spleen-associated lymphokines in mice immunized with FMD vaccineTo explore the effect of oral Danggui Buxue Tang on the expression levels of spleen-associated lymphokine m RNA in mice vaccinated with foot-and-mouth disease,24 Kunming mice were random Ly divided into 2 groups,12 in each group,namely Danggui Buxue Tang 0.5 g/m L dose group and In the control group(saline saline),all test mice were given 0.25 m L/d by intragastric administration for 5 days.Twenty-four hours after the last administration,the test mice were injected subcutaneously with 0.2 m L/type O of FMD vaccine in the groin,and Boost the immunization after 2 weeks.Mice were sacrificed by neck removal at the 3rd week after the second immunization,the spleen was harvested aseptically,the total RNA of the spleen was extracted by TRIZOL method,and the spleen-related lymphokines IL-4,IL-10,IFN-γand IL-21were analyzed by real-time fluorescence quantitative PCR m RNA expression level.The results showed that compared with the control group,oral Danggui Buxue Tang could significantly up-regulate the expression levels of IL-4 and IL-21 m RNA in the spleen(P<0.01),but had no significant effect on the expression levels of IL-10 and IFN-γm RNA in the spleen(P>0.05).It is suggested that oral Danggui Buxue Tang may increase the expression of IL-4 and IL-21 in the spleen of mice immunized with type O FMD vaccine to promote the production of specific antibody levels.3 Effect of oral administration of Danggui Buxue Tang on intestinal mucosal immunity in mice immunized with FMD disease vaccineTo investigate the effect of oral administration of Danggui Buxue Tang on the intestinal mucosal immunity of mice vaccinated with FMD vaccine,24 Kunming mice were random Ly divided into 2 groups,12 in each group,namely Danggui Buxue Tang 0.5 g/m L dose group and control group(Gastrointestinal saline);all mice tested were given 0.25 m L/d by intragastric administration for 5 days.Twenty-four hours after the last administration,the test mice were injected subcutaneously with 0.2 m L/type O of FMD vaccine in the groin,and boosted once every 2 weeks.Mice were sacrificed by neck removal at the 3rd week after the second immunization,the duodenum was aseptically removed,and the content of SIg A in the intestinal lavage fluid was detected by ELISA;the total RNA of the duodenum was extracted by the TRIZOL method,and real-time fluorescence quantitative PCR was used to detect twelve The m RNA expression levels of J-chain,p Ig R,BAFF,APRIL,IL-10 and IFN-γgenes in the intestinal tissue;prepare duodenal paraffin sections at the same time,count the number of epithelial lymphocytes(IEL)and immunohistochemistry by HE staining Count the area of Ig A~+cells.The results showed that compared with the control group,the SIg A content in the duodenum of mice was significantly increased after oral administration of Danggui Buxue Tang(P<0.01),the area of Ig A+cells was significantly increased(P<0.05),and the J-chain gene was significantly upregulated The expression level of m RNA(P<0.01),significantly up-regulated the m RNA expression levels of p Ig R,BAFF and IL-10 genes(P<0.05);there was no significant difference in the number of IEL(P>0.05).It is suggested that oral administration of Danggui Buxue Tan can increase the intestinal mucosal immunity of mice immunized with type O FMD vaccine.In summary,oral administration of Danggui Buxue Tang decoction can increase the levels of anti-FMD specific antibodies Ig G and its subclasses Ig G1 and Ig G2a in mouse serum,up-regulate the expression levels of splenic lymphocyte factor IL-4 and IL-21 m RNA,and promote SIg A secretion level in the small intestinal mucosa tissue increases Ig A~+cells in the intestinal mucosa,and at the same time enhances the m RNA expression levels of J-chain,p Ig R,BAFF,IL-10 genes in the duodenal tissue. |
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