Intervention Effect Of Water Extract Of Black Ginseng On Immunocompromised Mice

BackgroundBlack ginseng is prepared from white ginseng by steaming and drying 9 times repeatedly.During this process the color of white ginseng changes from white to red and finally to black,and the ginsenosides of white ginseng also changes to some rare ginsenosides,such as Rg3,Rh2,Rg2.The content of these rare ginsenosides increased greatly.Therefore,it is generally believed that black ginseng has more efficacy and stronger biological activity than white ginseng.In recent years,studies have found that the immune capacity of the body is closely related to the intestinal microecology.The intestinal microbiota can stimulate the immune function of the body,and the immune capacity of the body can affect the intestinal tract and its microbiota in turn.Previous studies in our laboratory have found that the intestinal tract of immunocompromised mice is prone to inflammatory infiltration,and mechanical barrier,biological barrier,immune barrier and chemical barrier of intestinal tract are all damaged.In this experiment,the immunocompromised mice model was used to verify whether the black ginseng processed in our laboratory could interfere the immunocompromise by regulating the immune function and repair the intestinal barrier by regulating the balance of intestinal microbiota.This experiment is of great significance for elaborating the pharmacodynamic mechanism of black ginseng and promoting the development and utilization of black ginseng products.ObjectivesDexamethasone was used to establish immunocompromised mice model.Black ginseng was processed and water extract of black ginseng was prepared to see the regulation effect of the water extract on immune function and the repair effect on intestinal barrier in the disease mice model.Methods1.Preparation of black ginseng and detection of ginsenoside content in black ginseng.Black ginseng was prepared by the method of "nine steaming and nine drying" with five years old wild ginseng as raw material.The contents of some rare ginsenosides were determined by HPLC and analyzed by external standard method.2.Establishment of immunocompromised mice model.BALB/c mice were selected as experimental animals and randomly divided into 4 groups(n=8): normal control group(NC),immunocompromised group(DEX),high concentration of black ginseng group(HB)and low concentration of black ginseng group(LB).The immunocompromised mice model was established by intraperitoneal injection of80mg/1000 g dexamethasone.Then,the black ginseng recovery groups were given black ginseng water extract by gavage every day,and the immunocompromised group was given the same volume of normal saline by gavage.The mice were sacrificed on the 30 th day,and the serum,spleen,thymus,intestine and other related tissues were collected and preserved for pathological,biochemical and immunological analysis.3.Detection of immunocompromise related indicators.The changes of thymus index and spleen index in mice were calculated.The levels of IL-6,IL-10 and TNF-α in serum of mice were determined by ELISA.The transformation ability of spleen cells was detected by MTT assay.4.Detection of intestinal related indicators.The expression of intestinal tight junction protein was detected by Western blotting.The expression of intestinal mucin(MUC2)was detected by immunohistochemistry.The changes of intestinal microbiota of mice were detected by DGGE and 16 Sr RNA gene sequencing.Results1.Compared with the normal control group,the body weight of the mice in the immunocompromised group was significantly reduced and the condition of the mice became worse;Thymus index and spleen index decreased.The expression of IL-6,IL-10 and TNF-α in serum decreased.The transformation ability of spleen cells was reduced.These results indicated that the immunocompromised mice model was established successfully.2.Compared with the immunocompromised mice,the body weight of the black ginseng recovery group increased;Thymus index and spleen index increased;The expression of IL-6,IL-10 and TNF-α in serum increased.The spleen cells transformation ability was enhanced.These results suggested that black ginseng can enhance the immune function.3.Compared with the immunocompromised mice,the expression of intestinal tight junction protein and mucin(MUC2)in the intestinal tract of mice in the black ginseng recovery group were increased,and the intestinal microbiota was more similar to that in the normal control group.This indicated that the water extract of black ginseng can alleviate the intestinal damage caused by immunocompromise.Conclusions1.The content of rare ginsenosides in black ginseng is higher than that in white ginseng.2.The water extract of black ginseng can improve the immune function of immunocompromised mice.3.Black ginseng water extract can repair the damage of intestinal mucosal epithelial barrier caused by immunocompromise,promote the balance of intestinal microbiota,and repair intestinal immunological barrier.