Effect Of Weifengning No.5 Recipe On Growth Of Human Coronary Artery Smooth Muscle Cells And Endothelial Cells In Vitro

BackgroundThe acute cardiovascular events caused by atherosclerotic cardiovascular disease(ASCVD) is the first killer seriously imperiling the health of people. Today percutaneous coronary intervention is becoming mature and perfect, and drug eluting stents(DES) has been taken as an important breakthrough in opening the occluded coronaries, however, there are still 20%-30% DES restenosis rate in placed DES and late stent thrombosis is the focus of great concern. Thus, it has significant meanings of exploring a new way to solve this problem. Professor Xian Wang thinks that the ASCVD, characterized by swift changes, rapid movement of clinical manifestation, similar to the wind syndrome of Traditional Chinese Medicine. Based on these theories, he proposed the hypothesis of "endogenous collateral wind" and created a formula called Luofengning-O(LFN-O) for the research and development of TCM eluting stents, which will provide an objective basis for the new theoiy of "endogenous collateral wind"PurposeIn order to provide a theoretical basis of the research and development of the stent LFN-0 complexes, we discussed the effect of LFN-0 complexes on the growth of human coronary artery smooth muscle cells(HCASMC) and human coronary artery endothelial cells(HCAEC) by cell culture, and then seeking out the appropriate ratio of paclitaxel and hirudin to maximize the inhibition of HCASMC and minimum the inhibition of HCAEC.Methods1. The culture of HCASMC and HCAEC:After the recoveiy, training, passages of HCASMC and HCAEC. we had established a stable cell experiment platform.2. Determined the effects of hirudin on the growth of HCASMC and HCAEC:By means of detecting the change of cells activity, we observed the state of normal cultured HCASMC and HCAEC which were treated with hirudin under different concentrations, and then determined the appropriate ratio of hirudin to maximize the inhibition of HCASMC and minimum the inhibition of HCAEC.3. Determined the effects of LFN-0 complexes on the growth of HCASMC and HCAEC:Detected the change of cells growth activity by MTT colorimetric method, and then observed the state of normal cultured HCASMC and HCAEC which were treated withLFN-0 complexes under different concentrations, and then determined the appropriate ratio of paclitaxel and hirudin to maximize the inhibition of HCASMC and minimum the inhibition of HCAECResults1. (1)The study on intervention of hirudin on HCASMC:Compared to the blank contrast group, low dose of hirudin(0.025-0.1mg/mL) was not apparently inhibit the growth of HCASMC(P>0.05); Medium dose and high dose of hirudin (0.2-6.25mg/mL) could obviously inhibited the growth of HCASMC(P<0.05).(2)The study on intervention of hirudin on HCAEC:Compared to the blank contrast group, 0.025-3.13mg/mL dose of hirudin was not apparently inhibit the growth of HCAEC(P>0.05); High dose of hirudin(6.25mg/mL)could obviously inhibited the growth of HCAEC(P<0.05), and 0.05-0.2mg/mL dose of hirudin can increase the growth of HCAEC.2. (1)The study on intervention of LFN-0 complexes on HCASMC:Compared to the blank contrast group, single paclitaxel and all dose of LFN-0 complexes could obviously inhibited the growth of HCASMC(P<0.05).(2)The study on intervention of LFN-0 complexes on HCAEC:Compared to the blank contrast group, the effect of single paclitaxel and all dose of LFN-0 complexes were apparently inhibited the growth of HCASMC(P<0.05); Besides, (1umol/L paclitaxel+0.39mg/mL hirudin)LFN-0 complexes group and (1umol/L paclitaxel+0.78mg/mL hirudin)LFN-0 complexes group could decrease the inhibition ratio of the growth of HCAEC compared with single paclitaxel group(P<0.05), all these proved the rationality of TCM compatibility to reduce the poison theory.Conclusion1. Low dose of hirudin is not apparently inhibit the growth of HCASMC, but medium dose and high dose of hirudin can obviously inhibited the growth of HCASMC.2.0.025-3.13mg/mL dose of hirudin are not apparently inhibit the growth of HCAEC, and 0.05-0.2mg/mL dose of hirudin can increase the growth of HCAEC3. For low dose of LFN-0 complexes (1 umol/L paclitaxel+0.39mg/mL hirudin) can maximize the inhibition of HCASMC and minimum the inhibition of HCAEC, we choose lumol/L paclitaxel matches 0.39mg/mL hirudin as our final ratio of LFN-0 complexes.