| Objective:In this study, through the fluorescent quantitative PCR method, we determined the expression of quantity of warm disease Yingfen rabbit model of intercellular adhesion molecule1(ICAM-1), vascular cell adhesion molecule1(VCAM1) and induced the expression of nitric oxide synthase (iNOS). To further verify from a more microscopic point of view of Yingfen, we have the pathological changes of blood stasis syndrome. At the same time, according to the main efficacy of drugs, the splitting formula of Qingying Decoction for Yin diathermy as the main function and Cooling blood stasis as the main efficacy of the drug group. Then we proposed the decomposed recipes research to further study the mechanism of pathological changes of Qingying Decoction to improve warm disease Yingfen animal model the syndrome of blood stasis. Therefore, we could disproof the pathogenic mechanism of blood stasis lesions of Yingfen. Meanwhile, we also discussed the effect of Qingying Decoction, its splitting formula on the blood stasis lesions of Yingfen and the mechanism of action from the preliminary.Methods:50New Zealand rabbits were randomly divided into the following five groups, i. e., Normal control group (group A), Model group (group B), Qingying Decoction group (group C), Cooling blood stasis group (group D) and Nourishing Yin diathermy group (group E), with10rabbits in each group. Except the Normal control group, each animal with a dexamethasone of2.5mg/kg daily injection to hind leg muscle. We made a continuous administration for seven days. The control group uses the same physiological saline injection on hind leg muscle. After the third day, except the Normal control group, the experimental animals were used traditional corresponding Chinese medicine5times than the equivalent dose of drug lavage for two times which are in the morning and at night. Normal control group and Model group were filled by the same amount of normal saline and lavage them for five consecutive days. In the evening of the7th day, each group of animals is banned on eating and drinking water. On the8th day, except the Normal control group, each group is injected a2.5ml/kg furosemide to the hind leg muscle. Two hours later, we made an injection of endotoxin of5ug to the experimental animals by ear marginal vein. Before the injection of LPS and after1,2,3,4,5hours, we measured the rectal temperature of each animal and drew an average temperature diagram of the reaction curve. Then we count heat peak net added value (△t) of each model animal and the changes of5hours heating index (TRI5). Five hours later, blood and viscera tissue samples are used to determine the hemorrheology, serum content of ICAM1, VCAM1and iNOS of each group. Through the fluorescence quantitative PCR method, we determined the expression of vascular intima intercellular adhesion molecule1(ICAM-1), vascular cell adhesion molecule1(VCAM1) and induce nitric oxide synthase (iNOS) in each rabbits groups. At the same time, the liver and lung tissue were stained with HE. Finally through the contrast to the Model group, we observe the Qingying Decoction and its splitting formula on blood stasis lesions of Yingfen and discussed its mechanism of action from the preliminary.Results:1. In terms of temperature:Compared with Normal control group, the heating net added value and5hours heat index of Yingfen Model group were significantly elevated, the statistics had extremely significant difference (P<0.01). It can be seen that the modeling of Yingfen is very successful. After traditional Chinese medicine therapy, the heating net added value and5hours heat index of TCM group A、group B and group C were significantly lower, compared with Model group, which has extremely significant difference (P<0.01, P<0.05). TCM group A reduced the rabbit Yingfen Model to the effect of temperature, which is the most obvious one. Group C of hypothermia close to TCM group A, there was no significant statistical difference (P>0.05). We compared group B with A and C, the statistics had extremely significant difference (P<0.01). The result shows that TCM group A and group C were significantly effect of hypothermia, and the role of TCM group B in terms of hypothermia is not obvious. It shows that the Whole Qinying Decoction has a good antifebrile effect and the Yin heating as the main efficacy plays a very important role in the abatement of fever. In hypothermia, the role of TCM group A and C are better than group B.2. In the aspect of hemorrheology:The hemorrheology related indexes of whole blood viscosity, reductive viscosity, plasma viscosity, erythrocyte deposited of Yingfen of the Model group is obviously enhanced, which is compared with Normal group. Its statistics had extremely significant difference (P<0.01). From the perspective of hemorrheology, full instructions of the blood of Yingfen animal model have obvious high adhesion, high pour-point and high poly changes of blood stasis. In different extent, TCM group A、group B and C improved the animal model of Yingfen group of hemorrheology indicators, especially in TCM group A, which the improvement effect is most obvious. In terms of the whole blood reductive viscosity (high, medium and low shear rate), plasma viscosity and erythrocyte deposited, Qingying Decoction group compared with Normal group, there was no significant-statistical difference (P>0.05). The reductive viscosity, low shear rate, red blood cells deposited values of TCM group TWO are close to the Qingying Decoction group (P>0.05). In whole blood viscosity, middle and low shear rate and reducing viscosity (high, medium and low shear rate, Yin diathermy group, and cool blood stasis group quite at leisure, the rest of the indicators compared with clear soup, activating blood stasis camp had significant difference (P<0.01or P<0.05). That Show Qingying Decoction and camp party down the two groups were able to improve the camp points the pathologic changes of the syndrome of blood stasis to different extent. But Qingying Decoction, cool blood stasis improved obviously.3. In terms of serum ICAM1, VCAM1, INOS:The levels of serum ICAM1, VCAM1and INOS in Model group were significantly increased, compared with the Normal group. It shows a statistically significant difference (P<0.01). After traditional Chinese medicine therapy, TCM group A and group B were significantly lower the model animal serum content of ICAM1, VCAM1and INOS, compared with Model group, which statistics have significant difference (P <0.01,P<0.05). Compared with the Normal group, the levels of serum ICAM1and the serum VCAM1in TCM group A were no significant statistical difference (P>0.05). Compared with the Normal group, there were no significant differences (P>0.05) in terms of the serum VCAM1of TCM group B. TCM group C has certainly improved the levels of serum ICAM-1and serum VCAM1of the model animal, but there was no effect on serum levels of INOS. The indicators of Qingying Decoction and its cooling blood stasis can all obviously reduce the levels of serum ICAM1, VCAM1and INOS of the model animal. Among them, the efficacy of Qingying Decoction is the most obvious one. The Yin diathermy Decoction mildly improves the levels of serum ICAM-1and serum VCAM1in the model animal, but the serum levels of INOS did not improve a lot. Qingying Decoction, cold blood stasis, nourishing Yin diathermy party can reduce the pathological model of Yinfen syndrome of blood stasis in terms of serum ICAM1, VCAM1and INOS and The Whole Qingying Decoction effect is most obvious.4. In terms of the expression quantity of vascular endothelial ICAM1, VCAM1, INOS:The expression level of Model group was significantly increased, compared with normal group, and the statistics had extremely significant difference (P<0.01). The neointimal ICAM1, VCAM1and INOS expression of three TCM group are significantly lower, compared with Model group, which has statistically significant difference (P<0.01, P<0.05). The vascular intima ICAM1、VCAM1、INOS expression levels of TCM group one was close to Normal group and there was no significant statistical difference (P>0.05). The vascular intima ICAM1、INOS expression levels of TCM group two was close to TCM group one and there was no significant statistical difference (P>0.05). Qingying Decoction, cold blood stasis, nourishing Yin diathermy party can reduce the pathological model of Yinfen syndrome of the expression of vascular intima ICAM1, VCAM1and INOS and The Whole Qingying Decoction effect is most obvious.Conclusions:1. The body temperature, hemorrheology indicators, serum ICAM1, VCAM1and INOS content of warm disease Yingfen rabbit model were significantly higher than the Normal group and the indicators of vascular endothelial ICAM1, VCAM1and INOS expression levels were significantly increased. On one hand, it confirmed that modeling the Yingfen animal model is successfully. On the other hand, it explained the pathologic changes of the syndrome and its blood stasis in terms of the perspective of more microscopic.2. The Whole Qingying Decoction can improve the symptoms of animal model, which can significantly reduce the temperature and hemorrheology indicators. Meanwhile it could lower the blood stasis serum ICAM1, VCAM1and INOS content and changes the vascular endothelial ICAM1, VCAM1, INOS expression level of Yingfen pathological model of syndrome of blood stasis pathologic. It could eventually improve the syndrome of blood stasis pathologic of Yingfen syndrome.3. the splitting formula of Yin diathermy drug and cool blood stasis in Qingying Decoction can improve the symptom of the camp points syndrome animal model, reduce animal models of body temperature, blood flow to improve sales points in the different degree certificate model degeneration and serum ICAM1, VCAM1, INOS content and vascular endothelial ICAM1, VCAM1, INOS expression level, eventually improve the yinfeng syndrome of blood stasis pathologic changes. The result proves the the two main factor of yinfeng for the formation of syndrome of blood stasis heat flourishing and battalion Yin loss, This and recorded in ancient books "blood heat is torn into pieces" and "the dragon’s fire burning the blood line the lag" is consistent. So for the business points"? of syndrome of blood stasis pathology treatment can not pay attention to cool the blood circulation, also should pay attention to nourishing Yin diathermy drug use.4. Compare with cooling blood stasis, the splitting formula of Yin diathermy drug group in Qingying Decoction, which plays the main function of drug group, has obvious advantages in hypothermia. But in terms of the improvement of then related parameters in blood stasis, the cold blood scattered stasis have an advantage of Yin diathermy. But the splitting formula of Yin diathermy medicine and cool blood stasis drugs are not as good as the general effect of Qingying Decoction all parties, The results indicated that cooling blood stasis, the splitting formula of Yin diathermy drug group in Qingying Decoction All the studies and has different, but both the role of the drug is complementary and indivisible, the pure camp is obviously better than the soup the overall effect of their individual role of party drugs, Qingying Decoction all side effect is not a simple Yin diathermy medicine addition cool blood scattered stasis drugs function and simple drug group are difficult to achieve the best effect of treatment, suggesting that Qingying Decoction formula compatibility with rationality and scientific nature. That Yin diathermy and cool blood scattered stasis two treatment does not replace the whole party and can not replace each other, two treatment compatibility with treatment on the synergy effect, checks and optimization. Also proved that the Chinese traditional medicine prescription is targeted at specific etiology and pathogenesis of the disease, from the law, in teaching, forming principle, method, square, the organic unity of the medicine.5. The mechanism of Qingying Decoction improves the blood stasis pathology of Yingfen, which may be caused by inhibiting the expression level of vascular endothelial ICAM1, VCAM1and INOS in order to lower the content of serum ICAM1, VCAM1and INOS. They could improve abnormal changes in hemorrheology of Yingfen animal model, which showed thick, sticky, coagulation and assembling. It eventually improves the syndrome of blood stasis pathology of Yingfen animal model. To dredge collaterals, guarantee the qi and blood circulation, improve the camp points syndrome of blood stasis pathology. It could also be Qingying Decoction the first druge use, truncation attempt, eventually stop the pathogenic heat to the zang-fu organs and the development to the blood the stages one of the mechanism of action of lesion severity. |
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