| Pidotimod is a synthetic substance with important immunomodulatory properties both in vitro and in vivo. Pidotimod is able to activate natural killer cell, enhance the phagocytic function of macrophage and neutrophilic granulocyte, promote the proliferation of lymphocyte induced by mitogens, and increase the production of cytokines such as Interleukin 2 andγ-interferon. Pidotimod is often utilized together with antibiotics and antivirotics to cure infectious diseases. Pidotimod is mostly used as a preparationsfororaluse, however, under some circumstances, it is necessary to make pidotimod into injection in order to speed up the absorption and increase its bioavailability. In this reaseach, the immunological activity of Pidotimod for injection has been studied.In chapter one, the effect of Pidotimod for injection on humoral immunity function and immune organs index of Balb/c mice was studied. In this experiment, 60 Balb/c mice were used, 36 of which were injected into Pidotimod with different doeses, 12 of which were injected into normal saline as negative control, and the rest of the mice were intragastrically perfused with Levamisole as positive control. All of the mice were doesd one time per day and were dosed for 14 times altogether. After 10 days of treatment, half of the mice were intraperitoneally injected with cyclophosphamide 80 mg·kg-1 one time per day for 4 times to establish the immunosuppressive model. On the 10th day of the treatment, all of the mice were intraperitoneally injected with SRBC. After the treatment, the serum hemolysin formations were performed by HC50 assay; Weights of thymuses and spleens of mice were measured. Result:The HC50 of each dosage of Pidotimod on normal mice was 158.044±1.669, 157.413±6.647, 157.571±3.919, respectively, there was no significant difference between each dosage of Pidotimod group and negative control group. The Thymus Index of each dosage of Pidotimod on normal mice was 2.469±0.267, 2.527±0.260, 2.590±0.277, respectively, there was significant difference between each dosage of Pidotimod group and negative control group. The spleen Index of each dosage of Pidotimod on normal mice was 5.355±0.779, 5.774±0.466, 5.690±0.676, respectively, there was significant difference between each dosage of Pidotimod group and negative control group; The HC50 of each dosage of Pidotimod on immunosuppressed mice was 8.044±1.304, 8.517±1.633, 7.939±1.045, respectively, there was no significant difference between each dosage of Pidotimod group and negative control group. The Thymus Index of each dosage of Pidotimod on immunosuppressed mice was 1.377±0.291, 1.676±0.301, 1.622±0.331, respectively, there were significant differences between medium and high does of Pidotimod groups and negative control group. The spleen Index of each dosage of Pidotimod on immunosuppressed mice was 3.139±0.544, 3.402±0.374, 3.593±0.501, respectively, there were significant differences between medium and high does of Pidotimod groups and negative control group. According to the result, Pidotimod for injection is not able to promote the formation of serum haemolysis in Balb/c mice; Pidotimod for injection can promote Thymus and Spleen Index of Balb/c mice.In chapter two, MTT essay was conducted to study the effect of Pidotimod for injection on spleen lymphocytes proliferation. In this experiment, 60 Balb/c mice were used, 36 of which were injected into Pidotimod with different doeses, 12 of which were injected into normal saline as negative control, and the rest of the mice were intragastrically perfused with Levamisole as positive control. All of the mice were doesd one time per day and were dosed for 14 times altogether. After 10 days of treatment, half of the mice were intraperitoneally injected with cyclophosphamide 80 mg·kg-1 one time per day for 4 times to establish the immunosuppressive model. After the treatment, spleen cells suspention was prepared and added into 96 wells culture plates, then the Con-A or LPS solution was added. After 48 hours of incubation in 37℃and 5% CO2, MTT essay was performed to evaluate the proliferation of spleen lymphocytes. Result: Under the inducement of Con-A,the OD value of each dosage of Pidotimod on normal mice was 0.890±0.167, 0.947±0.135, 1.120±0.097, respectively, there was significant difference between each dosage of Pidotimod group and negative control group(P<0.05, P<0.01, P<0.001). The OD value of each dosage of Pidotimod on immunosuppressed mice was 0.499±0.067, 0.524±0.043, 0.683±0.099, respectively, There was significant difference between each dosage of Pidotimod group and negative control group (P<0.001); Under the inducement of LPS, the OD value of each dosage of Pidotimod on normal mice was 0.545±0.044, 0.542±0.064, 0.714±0.106, respectively, There was significant difference between each dosage of Pidotimod group and negative control group P<0.001). The OD value of each dosage of Pidotimod on immunosuppressed mice was 0.277±0.037, 0.296±0.036, 0.298±0.025, respectively, There was significant difference between each dosage of Pidotimod group and negative control group(P<0.01, P<0.01, P<0.001). According to the result, Pidotimod for injection is able to promote the proliferation of spleen lymphocytes in Balb/c mice.In chapter three, neutral red colorimetry was used to study the effect of Pidotimod for injection on macrophage phagocytic function. In this experiment, 60 Balb/c mice were used, 36 of which were injected into Pidotimod with different doeses, 12 of which were injected into normal saline as negative control, and the rest of the mice were intragastrically perfused with Levamisole as positive control. All of the mice were doesd one time per day and were dosed for 14 times altogether. After 10 days of treatment, half of the mice were intraperitoneally injected with cyclophosphamide 80 mg·kg-1 one time per day for 4 times to establish the immunosuppressive model. After the treatment, peritoneal macrophages suspention was prepared and added into 96 wells culture plates, then neutral red solution was added. After 4 hours of incubation in 37℃and 5% CO2, colorimetry was used to evaluate the macrophage phagocytic function. Result: The OD value of each dosage of Pidotimod on normal mice was 0.516±0.047, 0.596±0.076, 0.968±0.271, respectively. There was significant difference between medium- dosage and high- dosage of Pidotimod group and negative control group (P<0.05, P<0.01). The OD value of each dosage of Pidotimod on immunosuppressed mice was 0.353±0.069, 0.421±0.054, 0.418±0.053, respectively. There was significant difference between each dosage of Pidotimod group and negative control group (P<0.05, P<0.01, P<0.05). According to the result, Pidotimod for injection is able to promote the Macrophage Phagocytic Function of Balb/c Mice. |
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