Effects Of1,25-dihydroxy Vitamin D₃on Proliferation Of CFs And TGF-β1in Rats Induced By AVP

Objective：To investigate the effect of1,25-dihydroxy vitamin D3（1,25D）on theproliferation of cardiac fibroblasts(CFs) induced by Argipressine(AVP) and itsrelationship with transforming growth factor-1(TGF-1) of cardiac fibroblasts(CFs)。Methods： I.Cell culture and processing:In vitro culture of rat cardiacfibroblasts were passaged and thawed by generalprocedure；II.Cell interference and culture： The CFs were divided into12holes and sent toculture。Each hole cell count5×103，and were divided into four groups：AVP group，VitD3group，AVP plus VitD3group and control group。AVP group including1holewas intervened with10－7mmol/L AVP；VitD3groups including A、B、C、D、E5holeswere joined the intervention with a concentration gradient of10-10,10-9,10-8,10-7,10-6mol/L for24hours；AVP plus VitD3groups including A、B、C、D、E5holes were intervened with10－7mmol/L AVP and VD3with a concentrationgradient of10-10,10-9,10-8,10-7,10-6mol/L；Control group including1hole has nointervention。After24hours interference and culture，return the supernatant andcollect cells。III.MTT assay was used to check absorbance value of the CFs in12holes of groups。IV.S period percentage in DNA synthesis of the rat CFs was measured by flowcytometry (FCM) in AVP plus VD3group、VD3group、AVP group and control group；V.Expression of TGF-β1protein of the rat CFs was measured by ELISA in AVP plusVD3group、VD3group、AVP group and control group，and expression of TGF-β1mRNA of the rat CFs was measured by RT-PCR in AVP plus VD3group、VD3group、AVP group and control group。 Result：1. Compared with control group(1.100±0.015)，absorbance value of CFs increasedsignificantly in AVP group(1.380±0.008）（P<0.05）；2. Absorbance value of CFs of AVP plus VD3groups relatively are A（1.345±0.005）、B（1.247±0.014）、C(1.198±0.011)、D(1.159±0.010)、E（1.115±0.013）, and eachgroup are concentration dependent(P<0.05)；The E hole has the lowest absorbancevalue（P<0.05）; Absorbance value of CFs of AVP plus VD3groups were relativelylower than that of control group(1.100±0.015)，but were relatively higher than that ofAVP group(0.776±0.013）（P<0.05）；3. Absorbance value of CFs in rats of VD3groups relatively are A（0.956±0.005）、B（1.006±0.012）、C（1.052±0.004）、D（1.058±0.011）、E（1.037±0.013）,all lowerthan the control group(1.100±0.015)(P<0.05)。4. S period percentage（SPF）of CFs in rats in AVP group（35.2%）、AVP plus VD3group（29.69%）measured by flow cytometry (FCM) is significantly higher than thecontrol group（22.57%）（P<0.05）；S period percent age of CFs in rats in AVP plusVD3group（29.69%VS35.2%）decreased compared with AVP group（P<0.05）；AndS period percentage of CFs in rats in VD3plus AVP group decreased compared withthe control group（19.25%VS22.57%）（P<0.05）;5. Expression of TGF-β1mRNA of CFs in rats（2.8928±0.2050VS1.0001±0.0139）is significantly higher than the control group（P<0.05）；Expression of TGF-β1mRNAof CFs in VD3plus AVP group(the E hole)（1.895±0.122VS2.893±0.2050）decreased compared with AVP group（P<0.05）; Expression of TGF-β1mRNA of CFsin VD3group（0.493±0.027）pg/mg decreased compared with other groups(P<0.05);6. Expression of TGF-β1protein in AVP group（78.310±4.803）pg/mg and AVP plusVD3group （62.479±5.846）pg/mg is significantly higher than the control group（44.29577±7.863）pg/mg（P<0.05）；Expression of TGF-β1protein of CFs in AVPplus VD3group(the E hole)（62.479±5.846VS78.310±4.803）pg/mg decreasedcompared with AVP group（P<0.05）；And expression of TGF-β1protein of CFs in VD3group(the E hole)（20.578±3.619）pg/mg decreased compared with othergroups(P<0.05);Conclusion：1.AVP has a role in promoting proliferation of CFs。2.Within a certain concentration range，1,25-dihydroxy vitamin D3effectively inhibitsthe proliferation of CFs of CFs induced by AVP，which maybe related to thedownregulation of myocardial TGF-1mRNA expression and protein synthesis.