| Objective: To observe the effects of Peiyuan Huayu Decoction-containing serum on the proliferation and migration of human microvascular endothelial cells(HEMC-1)and the expression of matrix metalloproteinases MMP-2,MMP-9,and vascular endothelial growth factor VEGF The inhibitory mechanism of Peiyuan Huayu Fang on chronic subdural hematoma(CSDH)adventitial microvascular formation,and the possible anti-inflammatory effect of the mechanism,provide a basis for clinical Chinese medicine to treat CSDH and reduce the postoperative recurrence rate.Methods: In experiment one,30 8-week-old SPF male Wista rats were divided into 5groups by random number table method.Each group consisted of 6 rats.They were blank control group,Peiyuan Huayu Recipe 0.5h,1h,and 1.5h.,2h group.One week after the adaptive feeding,in addition to the blank group,Peiyuan Huayu Decoction was administered with gavage at 12 g / kg / d for 3 days,and rats of each group were taken 0.5h,1h,1.5h,and 2h after the third day.Drug-containing serum.The sera of each sample taken from each group were identified by ligustrazine using high-performance liquid chromatography to verify whether the medicine-containing serum of Peiyuan Huayu Recipe was successfully prepared.In experiment two,after the serum containing Peiyuan Huayu Recipe was successfully prepared,it was temporarily stored in a refrigerator at-20 °C for later use.Human microvascular endothelial cells(HMEC-1)were purchased,cultured in endothelial cell-specific culture medium in a 5% CO2,37 ° C incubator,and plated in logarithmic phase.Cells were intervened with Peiyuan Huayu drug-containing serum and blank serum.To conduct cell proliferation experiments(cck-8),cell scratch experiments,and in vitro angiogenesis experiments to evaluate the effects of Peiyuan Huayu drug-containing serum on cell proliferation,migration,and angiogenesis.Western blot was used to determine VEGF,Expression of MMP-2 and MMP-9 proteins.To evaluate the expression of MMP-2 and MMP-9 in HMEC-1 before and after treatment,and to evaluate the effect of Peiyuan Huayu drug-containing serum on VEGF expression.Collect and organize experimental data for classification and summary analysis.Results: The results of experiment one suggest that the content of medicated serumligustrazine in the group 1.5h after the last gavage is higher than that in other groups,which proves that the preparation of medicated serum of Peiyuan Huayu Recipe was successfully prepared.In experiment two,the results of cell proliferation experiment found Compared with the blank group and the control group,the Peiyuan Huayu Decoction-containing serum group showed an inhibitory effect on cell growth,and the lowest OD value was found at a serum concentration of 15%.The results of cell scratch healing experiments showed that Peiyuan Huayu Decoction The 24 h migration distance of the drug-containing serum group was shorter than that of the blank serum group.In the in vitro angiogenesis experiment,after the Pei Yuan Huayu drug-containing serum intervened the cells for 3 hours,there was no significant difference in the tube formation area and number of lumens in the three groups.The number of lumens and the area of ??lumens were smaller than the blank group.In Western blot experiments,the MMP-2 and MMP-9of Peiyuan Huayu Decoction-containing serum were slightly lower than those of the blank serum group,and the expression of VEGF protein was much lower than that of the blank serum group.Conclusion: When preparing medicated serum,the content of ligustrazine in medicated serum collected from the abdominal aorta 1.5 hours after the last gavage was high;Peiyuan Huayu Fang medicated serum can inhibit the proliferation and migration of HMEC-1 cells and inhibit The formation of new blood vessels reduces the expression of VEGF,MMP-2,and MMP-9 proteins in HMEC-1 cells. |
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