| Objective: Antibody blocking method has applied to treat human endotoxemia for many years. However, because of high preparation cost of anti-endotoxin serum,which has not yet been used in veterinary clinical treatment. The aim of this study was to prepare endotoxin IgG antibody, and provide experimental and theoretical basis for the clinical treatment of endotoxemia.Methods: Canine feces were collected for isolation of E.coli, endotoxin were extracted with trichloroacetic acid and identified its activity by limulus reagent. Then injected intramuscularly to canine, 2 weeks later, the titer of serum was up to 1:64,injected again. The serum was taking from forearm venous blood by separating, then purified by caprylic acid ammonium sulfate, the supernatant of serum is IgG antibody.A total of 36 Kunming mouse were randomly divided into three groups: control group,endotoxemia group and serum treatment group. Specific IgG antibody was intraperitoneal injected 30 min after the subcutaneous injection of endotoxin at the concentration of 10 mg/kg. The endotoxemia group was administrated with same quantity of physiology brine. IgG antibody of LPS was intramuscularly administrated continuously for 7 days, once a day. After medication, mouse body temperature were measured by electronic thermometer after 1~7 d and comparing the changes of body temperature between treatment group and model group; blood routine were also determined and compared by blood cell analyzer after 1, 3, 7 d; The levels of TNF-a and IL-6 in serum of each group were measured by ELISA; Finally, pathological changes of liver and kidney were observed, what’s more, the above organs were fixed by neutral formalin and used to HE staining. The pathological changes of the liver and kidney tissue were observed under light microscope.Results:1. Changes of temperature: Temperature of positive control group was significantly increased(P<0.01), but serum treatment group were significantly decreased(P<0.01) compared with control group.2. Changes of blood routine examination: WBC and LYM of positive control group were significantly decreased(P<0.01) compared with the control group. Serum treatment group were all significantly increased(P<0.01), however, the changes of GRAN were the opposite.3. Changes of TNF-α and IL-6: Content of TNF-α and IL-6 in control group were332.7 ± 10.2 ng/L and 114.5 ± 4.06 ng/L, respectively, and positive control group were 460.6 ± 11.8 ng/L and 189.5 ± 20.9 ng/L, but serum treatment group were367.6 ± 7.6 ng/L and 128.9 ± 7.8 ng/L. The levels of TNF-α and IL-6 in serum treatment group significantly decreased compare to positive control group(P<0.01).4. Pothological examination: Compared with the positive control group, the treatment group was obviously better, liver cell cord arranged rule, part of the nucleus volume returned to normal; renal tubular epithelial cells arranged in neat and the infiltration of inflammatory cells was significantly reduced in the interstitial.Conclusion: Endotoxin serum has certain antagonistic effects on endotoximia through decreasing the number of inflammatory cells and the degree of inflammatory cell infiltration, and reducing the content of TNF-α and IL-6, which contributes to liver and kidney injury caused by endotoxin. |
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