| In currently veterinary clinic, gram-negative bacterial infectious disease of livestock occupies considerable proportion, such as E.coli and Salmonella. Endotoxin is released from the gram-negative bacteria and enters the blood circulation, causing the endotoxin blood disease (endotoxemia), and then inducing the animal body to appear in endotoxic shock, systemic inflammatory response syndrome (SIS), disseminated intravascular coagulation (DIC), which can affect the growth performance of livestock, thus influence the economic value of domestic animals. The inflammatory reaction is closely related to the coagulation/anticoagulant reaction, it may promote the coagulation reaction; the coagulation reaction may aggravate the inflammation reaction:the anticoagulant reaction also can resist the inflammation reaction. Xiang-Qi-Tang (XQT) is a traditional Chinese herbal compound which is composed of Rhizoma Cyperi. Astragalus membranaceus. Andrographis paniculatawith 4:3:3 as ratio. Previous studies have shown that it can reduce the expression of TNF-α and soluble endothelial protein C receptor (sEPCR) in chickens serum challenged by pathogenic Escherichia coli and has the function of anti-inflammation and anticoagulation. This study take the endotoxemia mice as a model to further research the effect of XQT, Rhizoma Cyperi. Astragalus membranaceus and Andrographis paniculata on expression of coagulation factors in mice with endotoxemia. The main methods and results are as follow:1. To establish the model of mice with endotoxemia:The sterile saline solution was given to the mice in blank group by intrapeitoneal injection at 0.1 ml/10g body weight, and LPS (0.25 mg/ml), LPS (0.5 mg/ml), LPS (1.0 mg/ml) were given to the mice in low dose group, middle dose group and high dose group by intrapeitoneal injection at 0.1 ml/lOg body weight. The results showed that the mice in each model group were curled up, flocked together, closed eyes. The temperature of mice was declined, the eyes turned red and moist. The mice had stool thin and other symptoms by injecting after 1 h. With the increasing of LPS concentration, the degree of feces formation and loose fur were increasing. The mice in low dose group and in middle dose group returned to normal after 24 h, the mice in high dose group returned to normal after 36 h. The expression of IL-6, ICAM-1, VCAM-1 were increased and the expression of EPCR was significantly decrease in each model group. The higher concentration of LPS was, the less expression of EPCR was. By comparing the expression of IL-6, ICAM-1, VCAM-1 and EPCR, the concentration of LPS solution in the mice model of endotoxemia was determined to be 0.5 mg/ml.2. Observation of XQT and its Chinese herbal medicine on the mental state of mice with endotoxemia:The sterile saline solution was used to treat the mice by intragastric administration in blank group and endotoxin model group at 0.1 ml/lOg body weight,1 time/d, used for 7 days. The Rhizoma Cyperi(0.1 g/ml), Astragalus membranaceus (0.75 g/ml), Andrographis paniculata (0.75 g/ml) and XQT (0.25 g/ml) were administered to the mice by intragastric administration in Xiang-Fu group, Huang-Qi group, Chuan-Xinlian group and Xiang-Qi-Tang group at 0.1 ml/lOg body weight,1 time/d, used for 7 days. The sterile saline solution was treated to the mice in dexamethasone group at 0.1 ml/10g body weight,1 time/d, used for 4 days and changed with the dexamethasone solution (0.2 mg/ml) on the fifth day, used for 3 days. The eighth day, the sterile saline was given to the mice in blank group by intrapeitoneal injection at 0.1 ml/lOg body weight, and LPS (0.5 mg/ml) was given to the mice in the other else groups by intrapeitoneal injection at 0.1 ml/10g body weight. The results showed that the mice in each model group were curled up, flocked together, closed eyes. The temperature of mice was declined, the eyes turned red and moist. The mice had stool thin and other symptoms by injecting after 1 h. They returned to normal after 24 h. The mice in treatment groups appeared curled up. flocked together, the feces moist but visible by injecting LPS within 12h, they returned to normal after 12 h. Rhizoma Cyperi, Astragalus membranaceus, Andrographis paniculataand and XQT can treat the mice with endotoxemia’s temperature in normal.3. Study on the effect of XQT and its Chinese herbal medicine on blood coagulation factors in mice with endotoxemia:Detected the blood coagulation factors in mice with endotoxemia by Western Blot, we found that the expression of sAPP. TF and sEPCR were significantly increased and the expression of tPA was significantly decreased. Rhizoma Cyperi, Astragalus membranaceus, Andrographis paniculataand and XQT could reduce the expression of sAPP. TF and sEPCR in different degree, while there was no decline in the expression of tPA, which pay a role in protecting the vascular endothelium. protein C anticoagulant pathway and fibrinolytic system.4. Study on the action mechanism of XQT and its Chinese herbal medicine on the mice with endotoxemia:The protein expression of AD AM17, MAPKs and PKC delta were significantly increased in mice with endotoxemia detected by Western Blot. Rhizoma Cyperi, Astragalus membranaceus, Andrographis paniculataand and XQT could reduce the expression of ADAM17. MAPKs and PKC delta in different degree. The results showed that XQT and its Chinese herbal medicine could reduce the expression of MAPKs and PKC delta on the blood vessels, to inhibit the activation of ADAM 17. and then made the shedding of EPCR, it is also pay a role in protecting the vascular endothelium and enhancing the function of anticoagulation. |
