[Effects of Ginseng and Sanchi Compositions on the Ras signal transduction pathway of myocardial ischemic rats].

OBJECTIVE: To study the effects of Ginseng and Sanchi Compositions (GSC) on the protein and mRNA expressions of Ras, extracellular signal-regulated Kinase1/2 (ERK1/2), and C-Raf-1 of ischemic myocardium of rats with acute myocardial infarction (AMI). METHODS: By adopting myocardial ischemia Wistar rat model with the ligation of the left anterior descending coronary artery, the normal control group, the sham-operation group, the model group, the Betaloc group, the high and low dose GSC group were set up. The protein expressions of Ras, C-Raf-1, ERK1/2,and phosphor-ERK1/2, p-ERK1/2 (p-ERK1/2) were detected by Western blot. The mRNA expressions of Ras, C-Raf-1, ERK1, and ERK2 were detected using Real-time PCR. RESULTS: The protein expressions of Ras, c-Raf-1, and p-ERK1/2 and their mRNA expressions in the model group increased more than those in the normal control group and the sham-operation group (P < 0.05). The protein expressions of Ras, c-Raf-1, and p-ERK1/2 and their mRNA expressions in the high and low dose GSC groups and the Betaloc group were significantly higher than those in the model group (P < 0.05). The protein expressions of Ras, c-Raf-1, and p-ERK1/2 and their mRNA expressions increased more obviously in the high dose GSC group than in the low dose GSC group (P < 0.05). The ERK1/2 protein expression was not significantly different among all groups (P > 0.05). CONCLUSIONS: GSC could up-regulate the protein expressions and mRNA expressions of Ras, C-Raf-1, and p-ERK1/2. It suggested that GSC might promote the angiogenesis through Ras signal transduction pathway dose-dependently.

[Action target of ginseng and panax notoginseng extracts on angiogenesis signaling pathway].

OBJECTIVE: To investigate the action targets of ginseng (GS) and Panax notoginseng (PN), Chinese herbs for benefiting qi and activating blood circulation, on angiogenesis signaling pathway (VEGFR-2-Ras-MAPK) in human umbilical vein endothelial cells (HUVEC). METHODS: To block the signal pathway by turns, at the first, added the IC50 of VEGFR-2 inhibitor, SU5416, and detected its downstream signaling protein Ras, MAPK expression using Western Blot. Secondly, added the IC50 of the Ras inhibitor, FPP, and detected its downstream signaling protein MAPK expression. RESULTS: Compared with the control group, adding SU5416 made the Ras, MAPK expression significantly decreased (P < 0.05, P < 0.01); and adding FPP made the expression of MAPK significantly decreased (P < 0.01). Compared with the group treated by SU5416 alone, the expression of downstream signaling protein Ras, MAPK were significantly higher in the group treated by SU5416 plus GS and PN (P < 0.05, P < 0.01); same state also found in comparison MAPK expression between groups treated with FPP alone and FPP plus GS and PN (P < 0.05). CONCLUSION: The angiogenesis mechanism of GS and PN on HUVEC may be realized by increasing the protein expression of three key signals, VEGFR-2, Ras and MAPK, respectively.

[Effects of Radix Ginseng and Radix Notoginseng formula on expressions of vascular endothelial growth factor receptor-2 and hypoxia-inducible factor-1alpha in ischemic myocardium of rats with acute myocardial infarction].

OBJECTIVE: To investigate the effects of Radix Ginseng and Radix Notoginseng formula on expressions of vascular endothelial growth factor receptor-2 (VEGFR-2) and hypoxia-inducible factor-1alpha (HIF-1alpha) in ischemic myocardium of rats with acute myocardial infarction. METHODS: A total of 100 Wistar rats were randomly divided into normal control group, sham-operated group, untreated group, metoprolol (Betaloc) group, and high- and low-dose Radix Ginseng and Radix Notoginseng formula groups. Acute myocardial infarction was induced in the untreated group, Betaloc group, and high- and low-dose Radix Ginseng and Radix Notoginseng formula groups by left anterior descending coronary artery ligation. After 12-day treatment, microvessel density (MVD) in ischemic myocardium was detected by immunohistochemical method, while expressions of VEGFR-2 and HIF-1alpha proteins were detected by Western blotting, and expressions of VEGFR-2 and HIF-1alpha mRNAs were detected by real-time fluorescent quantitative polymerase chain reaction. RESULTS: MVD in the untreated group was increased significantly, higher than those in the normal control group and the sham-operated group (P<0.05) and lower than those in the high- and low-dose Radix Ginseng and Radix Notoginseng formula groups and Betaloc group (P<0.01). VEGFR-2 and HIF-1alpha protein and mRNA expressions in the untreated group were higher than those in the normal control group and the sham-operated group (P<0.05). VEGFR-2 and HIF-1alpha protein and mRNA expressions in the high- and low-dose Radix Ginseng and Radix Notoginseng formula groups and Betaloc group were higher than those in the untreated group (P<0.05). There was a significant difference between the high- and low-dose Radix Ginseng and Radix Notoginseng formula groups (P<0.05). CONCLUSION: Radix Ginseng and Radix Notoginseng extract can up-regulate the protein and mRNA expressions of VEGFR-2 and HIF-1alpha and increase MVD in ischemic myocardium to improve myocardial ischemia so as to promote the development of collateral circulation.

Triptolide inhibits the function of TNF-α in osteoblast differentiation by inhibiting the NF-κB signaling pathway.

Chronic inflammation often delays fracture healing or leads to bone nonunion. Effectively suppressing pathological inflammation is crucial for fracture healing or bone remodeling. Triptolide, which is a diterpenoid epoxide, is the major active component of the Thunder God Vine, Tripterygium wilfordii. The aim of the present study was to investigate the role of triptolide in osteoblast differentiation and explore the molecular mechanisms of triptolide in fracture healing. Alkaline phosphatase (ALP) activity was used to evaluate osteoblast differentiation. ALP activity was measured via histochemical staining and western blotting was used to determine the expression of factors associated with inflammation. C2C12 cells were initially treated with 200 ng/ml bone morphogenetic protein (BMP)-2 alone for 3 days, which caused a significant increase in ALP activity (P<0.01). However, treatment with tumor necrosis factor (TNF)-α significantly decreased the ALP activity (P<0.05). Notably, treatment with the chronic inflammatory cytokine TNF-α significantly decreased the effect of BMP-2 in C2C12 cells compared with BMP-2 treatment alone (P<0.01). C2C12 cells were treated with increasing concentrations of BMP-2 or TNF-α for 3 days. The results demonstrated that TNF-α treatment significantly inhibited BMP-2-induced osteoblast differentiation in a dose-dependent manner (P<0.01). The role of triptolide in BMP-2-induced osteoblast differentiation was also examined. Cells were treated with BMP-2, BMP-2 + TNF-α alone, or BMP2 + TNF-α with increasing concentrations of triptolide (4, 8 or 16 ng/ml). After 3 days, the results of ALP activity revealed that triptolide significantly reversed the TNF-α-associated inhibition of osteoblast differentiation (P<0.01). Western blotting analysis demonstrated that triptolide markedly inhibited the phosphorylation of nuclear factor-κB, therefore suppressing the effects of TNF-α. In summary, triptolide is able to reverse the TNF-α-associated suppression of osteoblast differentiation, suggesting that triptolide treatment may have a positive effect on bone remodeling and fracture repairing.