| Objective:Leading by the theory"biological superficial signs are the key problem of syndrome model"and the method"the design of syndrome model should follow such a train of thought: cause - biological superficial signs-syndromes-treatment", this study aims at discovering new methods to set up and to evaluate animal model of blood stasis syndrome by observing the changing rules of biological superficial signs and biological basic of blood stasis syndrome animal model.Method:1. Wistar rats injected by HMWD as blood stasis syndrome model were used in this study. The rat models, which is marked by Evans Blue, are observed the changing rules of biological superficial signs in different times and does, detected Evans Blue content of focal tissue, interrelated indexes of blood rheology, contents of IL-1β,IL-6β,TNF-α,TXB2,6-keto-PGF1αin blood plasma.2. Wistar rats injected by lipopolysaccharide (LPS) and Carrageenan (CAR) as blood stasis syndrome model were used in this study. The rat models, which is marked by Evans Blue, are observed the changing rules of biological superficial signs in different times and does, detected Evans Blue content of focal tissue, interrelated indexes of blood rheology, contents of IL-1β,IL-6β,TNF-α,TXB2,6-keto-PGF1αin blood plasma.3. ICR mice intraperitoneal injected by Injection Ligustrazine Hydrochloride or Fufang Danshen Diwan, after 3-day successive administration, injected by HMWD as the blood stasis syndrome model, marked by Evans Blue, the mice are observed the changing rules of biological superficial signs, detected Evans Blue content of focal tissue.Result1. The rats injected HMWD show significant biological superficial signs as the blood stasis syndrome. There are blue ecchymosis in ear concha and tongue. As dosage and time multiplication, blue ecchymosis aggravates. Compared with control group, EB contents in plasma of model group decrease,(P<0.01); EB contents in ear concha tissue extract increase,(P<0.01); Blood viscosity(shear rate 150/s,38/s,10/s,5/s), plasma viscosity, Erythrocyte aggregation index raise up, (P<0.01/P<0.05), red blood cell deformation index decrease, but the difference is not significant; content of TXB2 in plasma increase, 6-keto-PGF1αin plasma decrease, TXB2/6-keto-PGF1αincrease, (P<0.01);content of IL-lβ, IL-6βin plasma is different, but the difference is not significant, TNF-αin plasma increase(P<0.01/P<0.05).2. The rats injected LPS/CAR show significant biological superficial signs as the blood stasis syndrome. There are blue ecchymosis in ear concha and tongue. As dosage and time multiplication, blue ecchymosis aggravates. Compared with control groups, EB contents in plasma of model group decrease, (P<0.01); EB contents in ear concha tissue extract increase,(P<0.01); Blood viscosity(shear rate 150/s,38/s,10/s,5/s), plasma viscosity, Erythrocyte aggregation index raise up, (P<0.01/P<0.05), red blood cell deformation index decrease, but the difference is not significant; content of TXB2 in plasma increase, 6-keto-PGF1αin plasma decrease, TXB2/6-keto-PGF1αincrease increase, (P<0.01/P<0.05) ; contents of IL-1β, IL-6β, TNF-αin plasma increase compared to control groups, (P<0.01/P<0.05).3. The extent of blue ecchymosis in group which are treated by drug for invigorating blood circulation and eliminating stasis is lighter compared to model groups, the EB contents in ear concha tissue extract is as well as less than the model groups, (P<0.01).Conclusion1. The Wistar rat, which is injected HMWD/EB as a blood stasis syndrome model, degree of superficial signs of blood stasis has does-effect relationship, time-effect relationship with HMWD.2. The Wistar Rat injected HMWD as blood stasis syndrome model, blood become thick, sticking, agglomerate and concretionary. TXB2 increase,6-Keto-PGF1αdecrease, content of IL-lβ, IL-6βin plasma is different, but the difference is not significant; TNF-αhas a dependability with blood stasis.3. The Wistar rat, which is injected LPS/CAR as a blood stasis syndrome model and colored by EB, the degree of superficial signs of blood stasis has does-effect relationship, time-effect relationship with LPS.4. The Wistar Rat injected LPS/CAR as blood stasis model, blood become thick, sticking, agglomerate and concretionary. TXB2 increase, 6-Keto-PGF1αdecrease, content of IL-lβ, IL-6β, TNF-αin plasma has a dependability with blood stasis. It indicates that inflammation is the key pathogenesy of this syndrome of blood stasis animal model.5. Drug for invigorating blood circulation and eliminating stasis such as injection Ligustrazine Hydrochloride and Fufang Danshen Diwan has a significant therapeutic action for mice injected HMWD. It lightens the superficial signs of blood stasis and indicates EB can express biological superficial signs of blood stasis syndrome in some way.Leading by the theory"the design of syndrome model should follow such a train of thought: cause - biological superficial signs-syndromes-treatment", the study adopts Evans Blue as the stain of blood stasis model, observes the changing rules of biological superficial signs, investigate evaluate mode, and differences between different blood stasis model, and arrived at a fine experimental results. |
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