Study The Effect Of Mollugo Pentaphylla On The Proliferation And Migration And Collagen Synthesis Of SD Rat Aortic Smooth Muscle Cells In Vitro

Objective: To Study the effect of Mollugo pentaphylla on the proliferation , migration and collagen synthesis of SD rat aortic vascular smooth muscle cells .Materials and Methods: About 5 passages of vascular smooth muscle cells(VSMC) were utilized in experiment by primary culture.VSMC were randomly grouped as follows:①blank control group (containing 10%FBS);②solvent control group (containing 1%DMSO & 10%FBS) ;③( 1,3,9,27μmol/L ) Mollugo pentaphylla(adding 10%FBS after a 60-min incubation of Mollugo pentaphylla ). Therefore,the method of MTT was adopted to observe the effects of different concentrations of Mollugo pentaphylla on the proliferation of VSMC; the way of Cell Scraping was to observe the effects of different concentrations of Mollugo pentaphylla on the migration of VSMC; and the effects of different concentrations of Mollugo pentaphylla on the content of hydroxyproline could be confirmed by method of Colorimetric.RESULTS: (1) VSMC identification: The majority of cells turned out to be spindle-shaped or long spindle-shaped in the inverted phase contrast microscope. After cells were stained by theα-actin immunohistochemistry,the light microscope showed that cytoplasm had brown granules and staining was positive, contrary to that in contrasting group. So by using the optical microscope and the immune cell chemical process appraisal,the cultured cells can be confirmed to VSMC, with high purity too.(2)The effects of Mollugo Pentaphylla with different dose on the proliferation of VSMC confirmed by MTT: Compared with blank control group and solvent control group, the VSMC extinction OD value of low dose(1μmol/L) Mollugo Pentaphylla groups increased by 22.7% at 72h and MTT restraining ratio with no significant difference(P > 0.05); However, Compared with blank control group, Mollugo Pentaphylla groups of different dose (3, 9, 27μmol/L) had lower OD value. Especially, high dose Mollugo Pentaphylla groups displaied significantly proliferation inhibitory effects. The inhibition rate of drug groups of VSMC at 72h was 42.8%(P﹤0.01), 62.4%(P﹤0.01),69.7%(P﹤0.01) respectively. In short, MTT indicated that Mollugo Pentaphylla has the inhibitory effect of dose dependency on the proliferation of VSMC.(3)The effects of Mollugo Pentaphylla with different dose on the migration of VSMC confirmed by method of cell scrape: Compared with solvent control group, blank control group raised the distance of migration of the VSMC with no significant difference;Mollugo Pentaphylla groups of different dose (1,3,9,27μmol/L) reduced the distance of migration of the VSMC, and inhibitory rates of migration were 4.4%(P <0.05～0.01),12.4%(P <0.01),20.5%(P<0.01) and 33.7%(P<0.01) respectively, which showed that Mollugo pentaphylla could inhibit the VSMC migration. It also indicated that Mollugo Pentaphylla has the inhibitory effect of dose dependency on the migration of VSMC.(4)The effects of Mollugo Pentaphylla with different dose on the hydroxyproline synthesis of VSMC confirmed by colorimetry method: Compared with blank control group and solvent control group, supernate hydroxyproline of 1μmol/L Mollugo Pentaphylla group changed slightly without difference in statistics; Supernate hydroxyproline of Mollugo Pentaphylla groups of different dose (3,9,27μmol/L) showed less hydroxyproline sythesis than blank control group and had significant difference too. So it indicated that Mollugo Pentaphylla has the inhibitory effect of dose dependency on the hydroxyproline synthesis of VSMC.CONCLUSIONS: It can be concluded that Mollugo pentaphylla can inhibit the proliferation and migration and collagen synthesis of rat vascular smooth muscle cells. This research may provide the experimental strategy to Mollugo pentaphylla for treating Blood vessel hyperplasia diseases.