Modified Model Of Thromboangiitis Obliterans In Rat And Study Of The Therapeutical Effect With The Extract Of Salvia Multiorrhiza Bge

ObjectiveThe aim of this study was to develop a new model of thromboangiitis obliterans (TAO) in rats and new evaluation index of the model, and to test the effect of extract of Salvia multiorrhiza bge.f.alba on this model.Methods50 rats (Wistar), were randomly divided into sham-operated group (n=10), 2.0mgLA group(n=10), 1.5mgLA group(n=10), 1.0mgLA group(n=10), 0.5mgLA group(n=10). 20mg/ml, 15mg/ml, 10mg/ml, 5mg/ml of sodium laurate (LA) were used respectively in each group to create models. And 0.1ml of each dosage was injected into the right femoral artery of every rats in these groups. 0.1ml of normal saline (N.S) was [0]used in sham-operated group. was finished, The right limb of rats were carefully observed after the injection. The observations of the lesion progression that include the skin color, swelling, gangrene and mummification of the affected leg were lasted for 2 weeks. The extent of the lesions of the 1st day, the 4th day, the 7th day, the 14th day were ranked by the Grade Evaluation of the Gangrene and the Gangrene Index(GI) and were recorded in corresponding forms. In the fifteenth day, Low shear viscosity, plasma viscosity, erythrocyte sedimentation rate(ERS), hematocrit value, erythrocyte relative index, plasma fibrinogen were measured to assess the hemorheology in TAO model; H-E-staining and triad colour staining (Masson way) were used to assess the lesions of limb of the experimental rats and the number of thrombosis were counted ; We also measured microvessel density (MVD) to presume the potential of neovasculrization in ischemic tissues.To optimize the model of TAO in rats. 24 rats were randomly divided into 0.8mgLA group(n=10), 0.6mgLA group(n=10), 0.4mgLA group(n=4);The ways of observation were the same as above.The therapeutical effect of the extract of Salvia multiorrhiza bge. 40 rats were randomly divided into sham-operated group (n=8), model group (n=8), aqueous extract of Salvia miltiorrhiza bge group(SM-A group, n=8), aqueous extract of Salvia miltiorrhiza bge.f.alba group(SMA-A group, n=8), ethanolic extract of Salviae miltiorrhiza bge.f.alba group(SMA-E, n=8). The progression of the lesions was recorded by the gangrene index. The ways of observation for the rest were the same as above.Result1 The appearance time and strength of the lesion closely connected with the dose of SL in the experimental rats. The rats of 2.0mgSL group, 1.5mgSL group have more grievous lesion, which gangrene to the ankle-jiont of rats around the 4th day. The gangrene extended to knee-joint around the fourteenth day. The lesions of 1.0mgLA group and 0.8mgLA group have the stationary process. The gangrene of some rats extended the ankle-joint around the seventh day . The lesions of 1.0mgLA group and 0.8mgLA group were limited in the front of sole.In the 7th day and the 4th day after the build up of the model, The result of the Grade Evaluation of the Gangrene had no statistical difference of 0.8mgLA group, 1.0mgLA group, 1.5mgLA group, 2.0mgLA group. The GI of the 0.8 mg LA group was significantly lower than that of 2.0mgLA group, 1.5mgLA group, 1.0mgLA group in the first day, the seventh day, the fourthday(P<0.05).Low shear viscosity, plasma viscosity erythrocyte aggregation index, plasma fibrinogen of 0.8mgLA group, 1.0mgLA group, 1.5mgLA group, 2.0mgLA group were significantly higher than that of sham-operated group. The MVD of 0.6mgLA group, 0.8mgLA group had significantly high levels.2. In the 4th day, GI of SM-A group, SMA-A group, SMA-E group were lower than sham-operated group. Low shear viscosity,plasma viscosity,erythrocyte aggregation index,plasma fibrinogen of SM-A group, SMA-A group, SMA-E group were lower than that of model group(P<0.05). Compared with sham-operated group and model group, MVD of SMA-E group increased significantly(P<0.05). Conclusion1. Injecting of LA through femoral artery , which is simple and convenient, is used to make TAO model. This method has good repeatability. The most appropriate dosage is 0.8mg～1.0mg which is decided by analyze and optimization.2. New objective evaluation index—gangrene index was made, standard of determination by quantitation and count method of this index were built up.3.The ethanolic extract of Salviae miltiorrhiza bge.f.alba , and were briefly proved to have therapeutical effectiveness, among which the extract of salvia multiorrhiza bge had strongest effect.