The Comparison Of The Effects Between The Lonicera Extract From Water Solution And The Probiotics On The Allergic BALB/c Mice

Part one The Effect of the Lonicera Extract from Water Solution on the Peritoneal Macrophages of the Allergic Mice in Vitro【Objective】To investigate the effect of the Lonicera extract from water solution (Lonicera ext) on the peritoneal macrophages of the ovalbulmin- induced hypersensitively mice in vitro.【Methods】The peritoneal macrophages(PM) were obtained from the ovalbumin(OVA) induced immediate hypersensitively murine model. Then the PM were incubated by 50%,25%,12.5%,6.25%,3.125%,1.0625% Lonicera ext for 24h in vitro, the capacity on the macrophages engulfing neutral red were detected and the metabolic levels of the macrophages were studied by using MTT colorimetric method.【Result】The capacity of the macrophage engulfing neutral red and the metabolic levels of the PM incubated by 50%, 25%, 12.5% and 6.25% of the Lonicera ext were significantly increased. 【Conclusions】The phagocytotic capacity and metabolic levels of the PM obtained from OVA-induced allergic mice were promoted by the higher concentration of the Lonicera ext in vitro, suggesting the PM were activated.Part twoThe Comparison of the Effects between the Lonicera Extract from Water Solution and the Probiotics on the Allergic BALB/c Mice【Objective】To investigate the therapeutic effects of the Lonicera ext compared with the probiotics in the OVA-induced allergic BALB/c Mice.【Methods】Female BALB/c mice aged at 4-5 weeks fed on the OVA-free diet, were randomly divided into 5 groups by the intragastric administration after being sensitized and challenged with OVA including group LTH treated with 100%,group LTM with 50% of the Lonicera ext, group PT with the probiotics and group Ch treated with normal saline as the positive control. Meanwhile, the mice in group NS without sensitization and challenge were set as the negative control. (1) The tissues of small intestine were histologically examined by both HE and toludine blue stain. (2) The capacity of the peritoneal macrophages engulfing chicken red blood cells were studied by Wright stain. (3) The stool samples of mice were collected, diluted and cultured on specific selective agar plates to detect Bifidobacterium and Lactobacillus. Polymerase chain reaction method was used to identify bacterium genus. (4) The levels of the OVA-specific IgE and IgG2a in the serum of the mice were measured by ELISA. (5) The levels of total IgA and OVA-specific IgA in the intestinal mucus of the mice were determined by ELISA.【Result】OVA-induced allergic mice were treated with Lonicera ext and probiotics by the intragastric administration for 10 days: (1) The significant inflammatory reactions, such as the accumulation of the lymphocytes and plasmacytes in the lamina propria (LP) and the focal necrosis and shedding of the epithelial cells in the intestinal villi were inhibited. (2) The accumulated and degranulated mast cells in the LP were statistically reduced. (3) The percentage of the peritoneal macrophage phagocytosis and phagocytotic index were markedly improved. (4) The counts of Bifidobacterium and Lactobacillus in the stool samples of the sensitive mice taken with Lonicera ext and probiotics were raised, especially in the snestive mice treated by 100% Lonicera ext, and there were no differenece between group LTM and PT. (5) Both the levels of OVA-specific IgE (SIgE) of the serum in the allergic mice treated by Lonicera ext and probiotics were depressed and the SIgG2a were boosted, so that the ratios of SIgE/SIgG2a were statistically reduced. (6) The total IgA and OVA- specific IgA in the intestinal mucus in group LTH were higher than that group PT.【Conclusions】The present study first showed that sensitive mice treated with Lonicera ext were an improvement on the probiotics.(1) The accumulated and degranulated mast cells in LP were obviously inhibited,and the phagocytotic capacity of the peritoneal macrophage were promoted;(2) The counts of Bifidobacterium and Lactobacillus rised in the intestine of the allergic mice, especially in the snestive mice treated by 100% Lonicera ext;(3) Both the levels of OVA-specific IgE (SIgE) of the serum in the allergic mice treated by Lonicera ext and probiotics were depressed, the SIgG2a were boosted, and the ratios of SIgE/SIgG2a were statistically reduced. These results suggested that Lonicera ext and probiotics may recover the Th1/Th2 balance;(4) The total IgA and OVA-specific IgA in the intestinal mucus of the allergic mice were significantly augmented in the group LTH, suggesting that Lonicera ext may regulate the intestinal mucosal immunity of the allergic mice through promoting the clearance of harmful antigen and preventing the diffusion of inflammation;(5) Though there were no differenece between the group LTM and PT in the intestinal probiotics of the allergic mice, both the ratios of the SIgE/SIgG2a were depressed and the levels of the OVA-specific IgA were raised significantly in the group LTM ccompared with that in group PT. Our results showed that Lonicera ext may directly regulate immunological functions on allergic mice, and the effects of Lonicera ext on improving the intestinal fuctions and Th1/Th2 balance were better for allergic mice than that of the probiotics.(6) Our results suggested that the recovery of the immunological disequilibrium need a long time, because the improvement of intestinal symptoms was earlier than the recovery of the specific immunoglobulin. The further study needs to investigate whether the immunological disequilibrium recoveres completely and the intestinal mucosal immunoregulatory mechanisms treated by Lonicera ext and probitics of the allergic mice.