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Study of Protective Function Mechanism of Xintongshu Decoction (XTSD) in Treatment of Cardiac Muscle on JAK-STAT、SOCS Signal Pathway

Author: ChenChuTao
Tutor: HuangZhengDe
School: Hunan University of Traditional Chinese Medicine
Course: Chinese medical science
Keywords: model of blood-stasis syndrome plus myocardial ischemia-reperfusion injury (IRI) Xintongshu Decoction (XTSD) JAK-STAT signal pathway negative regulative system of SOCS hypoxia/reaeration group (H/R)
CLC: R285.5
Type: PhD thesis
Year: 2010
Downloads: 110
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Abstract


Objective:Models with blood-stasis syndrome and IRI were set. Based on pathomorphology, the expression of JAK-STAT signal pathway in models were explored; based on JAK-STAT signal pathway, pre-protective mechanism of Xintongshu Decoction in treatment of IRI in rats with blood stasis syndrome was studied.Methods:This research includes 3 parts as following:1.32 SD rats were divided into 4 groups randomly with 8 rats per group. They were the sham-operated group, blood-stasis syndrome group, IRI group, blood-stasis syndrome plus IRI group. Histomorphology of myocardium was observed by HE staining; the contents of Il-6 and IL-10 in JAK-STAT signal pathway were measured by ELISA; the protein expressions of JAK1, JAK2, STAT1 and STAT3 close related with myocardial tissue in the signal pathway were tested by Western Blot; the setting method of blood-stasis syndrome plus IRI model was studied.2.48 SD rats were divided into 6 groups randomly with 8 rats per group. They were the sham-operated group, blood-stasis syndrome plus sham-operated group, blood-stasis syndrome plus 30min IRI group, blood-stasis syndrome plus 60min IRI group, blood-stasis syndrome plus IRI group, blood-stasis syndrome plus 30min IRI with XTSD group, blood-stasis syndrome plus 60min IRI with XTSD group, histomorphology of myocardium was observed by HE staining; the contents of 11-6 and IL-10 in JAK-STAT signal pathway were measured by ELISA; the protein expressions of JAK1, JAK2, STAT1 and STAT3 in the signal pathway were tested by Western Blot法; the mRNA expressions of SOCS1, SOCS3 in myocardial tissue were measured by RT-PCR. The pre-protective mechanism of Xintongshu Decoction in treatment of IRI in rats with blood stasis syndrome based on the negative regulation of JAK-STAT and SOCS signal pathway was studied.3. By using 6-orifice plate, myocardial cells of primary newborn rats were cultured which were divided into 4 groups with 6repeated orifices per group: control group, hypoxia/reaeration group (H/R group), verapamil hydrochloride group (VH group) and XTSD group. The morphological changes of H/R group of myocardial cells were observed by Rhodamine staining. The protein expressions of JAK1, JAK2, STAT1 and STAT3 were tested by Western Blot. The protection effect of XTSD on IRI was proved by cells in vitro and the mechanism was related with the JAK-STAT signal pathway.Results:1. Compared with those in sham-operated group, QRS complex of ECG in blood-stasis syndrome group became higher and wider, J Point ascended, while pretreatment of XTSD could decrease the J point; By using HE staining, it could be seen that the edema of myocardial cells, fracture of muscle fiber, karyolysis or disappearance of a few myocardial cells and a small mount neutrophil infiltrations. The pretreatment of XTSD could reduce the symptoms mentioned above.2. Compared with those in sham-operated group, the level of hemorheology indexes and thromboxane B-2 (TXB2) increased obviously and the difference was remarkably significant (P<0.01). The level of 6-Keto-PGF1αand NO decreased obviously, and the difference was remarkably significant (P<0.01). Compared with those in blood-stasis syndrome group, the indices of hemorheology indexes and TXB2 in blood-stasis syndrome plus IRI group increased obviously and the difference was remarkably significant (P<0.01). The level of 6-keto-PGF1αand NO decreased obviously and the difference was remarkably significant (P<0.01).3. Compared with those in sham-operated group, the level of IL-6 and IL-10 in blood-stasis syndrome group increased obviously, and the difference was remarkably significant (P<0.01). Compared with those in blood-stasis syndrome group, the level of IL-6 and IL-10 in blood-stasis syndrome plus IRI group increased obviously and the difference was remarkably significant (P<0.01).4. Compared with those in sham-operated group, the level of JAK1, JAK2, STAT1 and STAT3 protein of myocardial tissue in blood-stasis syndrome group increased obviously, and the difference was remarkably significant (P<0.01). Compared with those in blood-stasis syndrome group, the level of JAK1, JAK2, STAT1 and STAT3 protein in blood-stasis syndrome plus IRI group increased obviously, and the difference was remarkably significant (P<0.01).5. Compared with those in blood-stasis syndrome plus 30 or 60 min IRI group, the expression of JAK1 and STATA protein in blood-stasis syndrome plus 30 or 60 min IRI with XTSD group decreased obviously, and the difference was remarkably significant (P<0.01). The expression of JAK2 and STAT3 protein increased obviously, and the difference was remarkably significant (P<0.01).6. Compared with those in control group, the morphology of the myocardial cells in H/R group was extremely irregular and with a large amount of fragments; the morphology of myocardial cells in VH group was regular with more fragments; the morphology of myocardial cells in XTSD group was regular with few fragments.7. Compared with those in H/R group, the expression of JAK1 and STATA protein in XTSD group decreased obviously, and the difference was remarkably significant (P<0.01). The expression of JAK2 and STAT3 protein increased obviously, and the difference was remarkably significant (P<0.01).Conclusion:The model of blood-stasis plus IRI of rats is more suitable to the pathogenesis of TCM and can supply the experimental objects more closed to clinic for the researches of the treatment on IRI with TCM. The protective mechanism of XTSD on the rats with blood-stasis syndrome plus IRI is related with JAK-STAT and SOCS signal transduction pathway. Experiment in vitro further proves that XTSD can do the protective effect on IRI by activating JAK-STAT signal transduction pathway.

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