| Objectives:Nowadays, the general public are most concerned about health. The goal is giving full play to all the body’s physiological function and to promote the physical and mental health. Therefore, study on the promotion of natural active substances for healthy physical activity and the toxic side effects has become a research hotspot in recent years. Because chronic non-communicable diseases, such as cardiovascular disease, cancer, diabetes are closely related to the occurrence of the body to produce excessive free radicals, reducing immune function, so the research for antioxidant, scavenging free radicals and enhance the immune function of natural active constituents has become an important part of functional foods.Rhodiola is a perennial herb, which has 90 kinds all around the world, but there are 73 species in China, Rhodiola are mainly distributed in Changbai Mountain. And Rhodiola has broad pharmacological functions, including antioxidant, antitumor, antiviral, regulate the nervous system function. For the demand is increasing, followed by excessive logging, the resources is lack. The main active components of Rhodiola are aglycone and tyrosol,free radical scavenging capacity of hydroxyl is much higher than Rhodiola. Reports show that the content of Rhodiola are able to be increased and tyrosol could be toreproduce by microbial fermentation method, cultivate the microorganism fermentation process,but it is too complicate. Another report show, not only by lactic acid bacteria fermentation liquid of Rhodiola, but with red ginseng extract and fermentation, fermentation has better effect, and improve the content of tyrosol and the ability of scavenging free radicals. In this study, with the use of active dry yeast, mixing the Rhodiola and Codonopsis lanceolata fermented raw materials together, we test the effects of two kinds of material mixed fermentation extracts on immune function in mice.Methods:1. In vitro serum pharmacology experiment:10 rats were divided into 5 groups, with 2 rats in each group, namely the blank group(Control), unfermented Rhodiola group (NF-R), non-fermented Codonopsis lanceolata group (NF-C), Rhodiola and Codonopsis lanceolata combined fermentation (F-R+C) group, Rhodiola and Codonopsis lanceolata crude drug combination group (NF-R+C). The control group was treated with physiological saline, other groups administered by different drug extract,0.1 ml/10g, continuous administration for 5 days, 1 hours after the last administration, sterile take eye blood, centrifuge at 3000r/min for 10 minutes, separation of serum, each with 56 ℃. inactivation of 30min. Using in the T, B lymphocyte proliferation and NK cell activity of experimental results.2. Experimental determination of immune function in vivo:80 Balb/c mice of clean grade, were randomly divided into 4 groups:blank control group (distilled water O0.1ml/10g), rhodiola and Codonopsis lanceolata mixed fermentation group (F-R+C, rhodiola and Codonopsis lanceolata mixed fermentation extract of 50mg/kg) and unfermented rhodiola and Codonopsis lanceolata mixed group (NF-R+C, rhodiola extracts of 50mg/kg) and PDT dispersible tablet solution group (PDT, positive control group,50mg/kg), with 20 rats in each group, one group with 10 rats in each group for the delayed type hypersensitivity (delayed type hypersensitivity DTH)experiment, the remaining 10 is only used for determination of the activity of mouse spleen lymphocyte proliferation and NK cell cytotoxicity. All the animals were orally given the corresponding test administered continuously for 30 days. Record the weight and calculate the thymus and spleen index, and determinate culture liquid of spleen interleukin-2(IL-2),interleukin-6(IL-6),interleukin-10(IL-10), interferon (IFN-γ), tumor necrosis factor(TNF-)by ELISA.Results:1. In vitro drug containing serum of experimental results show that:compared with the control group, rhodiola and Codonopsis lanceolata crude combined fermentation extract group and rhodiola and Codonopsis lanceolata crude mixed group can significantly increase ConA, LPS induced T and B lymphocyte proliferation and NK cell activity (P<0.01).2. Results of the body weight and organ index showed that PDT, rhodiola and Codonopsis lanceolata combined fermentation extract group (F-R+C) and unfermented group (NF-R+C). The body weight of mice compared with the control group, there was no significant difference between the three groups, and the thymus index were significantly higher than those in control group (P＜0.01), and the effects of Rhodiola and Codonopsis lanceolata combined fermentation extract group thymus index was significantly higher than that of unfermented group, the difference was statistically significant (P＜0.05). PDT group and rhodiola and Codonopsis lanceolata combined fermentation extract group spleen index increased significantly, compared with the control group, the difference was statistically significant (P<0.05), and rhodiola root and Codonopsis lanceolata combined fermentation extract group spleen index was significantly higher than that of unfermented group, the difference was statistically significant (P<0.05).3. In vivo effects of proliferation of spleen lymphocytes and NK cell activity in mice. The results showed that:rhodiola and Codonopsis lanceolata mixed group and T lymphocyte and B lymphocyte proliferation of unfermented group and the activity of NK cells in PDT group were significantly higher than those in the blank control group, (P<0.01), and the T lymphocyte and B lymphocyte proliferation and NK cell the activity of the rhodiola and Codonopsis lanceolata mixed group was significantly higher than that of unfermented group (P<0.05). Compared with PDT group, the activity of NK cells were significantly lower than that of PDT group, but the activity of NK cells in fermentation group and PDT group had no significant difference, indicate that after fermentation, the activity of NK cells to enhance immune function and positive drug control group.4. In vivo effects on delayed type hypersensitivity in mice showed that:group PDT, group of Rhodiola and mixed fermentation of Codonopsis lanceolata and unfermented group, ear swelling was significantly higher than that of control group (P＜0.01). Rhodiola and Codonopsis lanceolata mixed fermentation group of ear swelling was significantly higher than that of non-fermentation group, the difference was statistically significant (P＜0.01).5. Mouse spleen cell culture fluid interleukin-2 (IL-2) determination results that:compared with the control group, PDT can be significantly improved medium IL-2 of mouse spleen cells, the difference was statistically significant (P＜0.01). Compared with the control group, the content of F-R+C and NF-R+C group could significantly improve the mice spleen cell culture fluid IL-2, the difference was statistically significant (P＜0.01). Compared with NF-R+C group, the content of F-R+C group mice spleen cell culture fluid of IL-2 significantly increased, the difference was statistically significant (P＜0.01).6. Mouse spleen cell culture fluid interleukin-6 (IL-6) determination results that:compared with the control group, PDT can be significantly improved medium IL-6 of mouse spleen cells, the difference was statistically significant (P＜0.01). Compared with the control group, the content of F-R+C and NF-R+C group could significantly improve the mice spleen cell culture medium of IL-6, the difference was statistically significant (P＜0.01). Compared with NF-R+C group, the content of F-R+C group mice spleen cell culture fluid of IL-6 significantly increased, the difference was statistically significant (P<0.01).7. Mouse spleen cell culture fluid interleukin-10 (IL-10) determination results that: compared with the control group, PDT can be significantly improved medium IL-10 of mouse spleen cells, the difference was statistically significant (P<0.01). Compared with the control group, the content of F-R+C and NF-R+C group could significantly improve the mice spleen cell culture medium of IL-10, the difference was statistically significant (P＜0.01). Compared with NF-R+C group, F-R+C group, the content of mouse spleen cell culture fluid of IL-10 significantly increased, the difference was statistically significant (P<0.01).8. Mouse spleen cell culture liquid of interferon (IFN-y) determination results that: compared with the control group, PDT can be significantly improved medium IFN- gamma in mouse spleen cells, the difference was statistically significant (P<0.01). Compared with the control group, the content of F-R+C and NF-R+C group could significantly improve the mouse spleen cells by IFN-y, the difference was statistically significant (P＜0.01). Compared with NF-R+C group, F-R+C group, the content of mouse spleen cell culture fluid of IFN-y significantly increased, the difference was statistically significant (.P<0.01).9. Mouse spleen cells of tumor necrosis factor (TNF-a) determination results that: compared with the control group, the content of PDT can be significantly improved medium TNF-a mice spleen cells, the difference was statistically significant (P＜0.01). Compared with the control group, the content of F-R+C and NF-R+C group could significantly improve the mice spleen cell culture fluid of TNF-a, the difference was statistically significant (P＜0.01). Compared with NF-R+C group, the content of F-R+C group mice spleen cell culture fluid of TNF-a was significantly improved, the difference was statistically significant (P<0.01).Conclusions:1. Fermentated extract of mixed Rhodiola and Codonopsis lanceolata can significantly improve the immune function of mice.2. Rhodiola and Codonopsis lanceolata mixture by yeast fermentation, he enhancement of immunity function improved significantly. |
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