Left-handed Amlodipine On Spontaneously Hypertensive Rats Vascular Smooth Muscle Cell Proliferation And CaN Influence

Objective:Observation of spontaneously hypertensive rats (SHR) morphology andproliferation of vascular smooth muscle cells; Detection of SHR vascular smoothmuscle cells in the activity of calcineurin (CaN); Observe the calcium antagonist-left-handed amlodipine intervention of SHR inhibition of vascular smooth muscle cellproliferation, to explore the relationship with the CaN.Methods:Spontaneously hypertensive rats, male, weeks of9weeks, weight quality of190-210g, measuring the rat tail blood pressure: the roots of systolic blood pressure to170±12mmHg, diastolic blood pressure to98±7mmHg. Sterile for rat aorta,membrane preparation, subculture.Experiment is divided into the control group(group) with essential hypertension (eh), the experimental group (group A:10umol/L,group B:1.0umol/L, group C:0.1umol/L). Identification of VSMC byimmunohistochemical method, Inverted phase contrast microscope observation ofVSMC morphology, cell survival rate and growth curve, determined by MTT methodto measure VSMC proliferation and ELISA method CaN activity.Data analysis usingSPSS16.0statistical software processing.Results:1.The immunohistochemical method appraisal VSMC: Cell cytoplasm or coatedappeared pale yellow to dark brown dye, nonspecific staining and eliminate background, proved to be vascular smooth muscle cells.2.Inverted phase contrast microscope observation of VSMC morphology, draw2generation growth curve with trypan blue staining method to detect cell survival rate:Observed the original generation3days or so to see a number of organizations aroundthe block a cell,5-7days later, the proliferation of cell division into the logarithmicphase,8days later, near the tissue pieces for the first time in vascular smooth musclecells characteristic of "valley-peak" structure; Cell survival rate detection: live cellswere over96%, shows that high survival rate of cultured cells extend; Draw thegrowth curve, cell appears "incubation period-logarithmic phase-stagnant period"of growth pattern.3.Determined by MTT method to measure each VSMC proliferation and sinistralamlodipine intervention effects on its: Greenhouse-Geisser results show:Comparison between each time: F=7.387，P=0.000, With the extension of time, thecell has been on the rise; By the silhouette visible:5to6days of group C, N groupday cell proliferation peak; Comparison between group C and N group: F=3.678, P=0.000, which indicates that cell proliferation trend is different between two groups,VSMC proliferation in control group compared with model group showed a trend ofslow growth and that of model group multiplication effect is better than the controlgroup; The interaction of the treatment group and time: F=4.534，P=0.000,there isinteraction between high blood pressure and time, Interaction between high bloodpressure and time, they mutual influence cell proliferation, high doses of sinistralamlodipine abnormal VSMC proliferation inhibition of SHR significantly (P <0.05).4.The activity of ELISA method CaN change and sinistral amlodipine interventioneffect on its: N group CaN activity is significantly higher than group C (5.22±0.34vs. 2.81±0.18, P <0.05), indicating that SHR CaN activities; Group H CaN activity waslower than that in group N (3.98±0.13vs5.22±0.34, P<0.05), indicating that highdoses of sinistral amlodipine CaN reduce SHR CaN activity to a certain extent.Conclusions:1.Activities of in spontaneously hypertensive rats CaN prompt the enzymes involvedin vascular smooth muscle cell proliferation.2.Left-handed amlodipine by reducing the activity of CaN, to participate in theinhibition of vascular smooth muscle cell proliferation.