1、Adrenomedullin inhibits vascular calcification nDepartment of Physiology and Pathophysiology,Peking University Health Science CenternKey Laboratory of the Ministry of Education on Molecular CardiologynYong-Fen Qi2020/10/181Vascular calcificationHypertensionDiabetes Mellitus HyperlipemiaRenal Failure
2、 Aging MediaIntima Vascular calcification is believed to be an important risk factor of cardiovasculardiseasesAtherosclerosis2020/10/182Vasoactive peptides:Adrenomedullin?2020/10/183nAdrenomedullin(ADM)is a peptide capable of inducing a potent and long lasting hypotensive effect nADM is mainly synth
3、esized and secreted from vascular endothelial and smooth muscle cells(VSMC)nADM has been shown to have vasodilatory,hypotensive and growth regulating properties,nExogenous ADM or ADM gene delivery attenuated hypertension and protected against cardiovascular remodeling,but ADM gene knockout resulted
4、in higher blood pressure and cardiovascular abnormalities.nEndogenous ADM is an important factor in regulating cardiovascular and renal homoestasis,and is a potent cardio-reno-protective factor2020/10/184nreceptors bound with ADM were orphan L-1 and RDC-1 receptors nIn addition,another orphan recept
5、or named calcitonin receptor-like receptor(CRLR)could bind with ADM when receptor activity-modifying protein(RAMP)2 and/or RAMP3 existed on cellular membrane surface 2020/10/185Fig.1.(A)Calcification of multicellular nodules.von Kossa staining for calcification,showing positive staining as black/bro
6、wn areas within the main,large,nodular structure(original magnification 40).(B)Formation of multicellular nodules in culture phase contrast microscopy showing rat VSMCs(original magnification 20).AB2020/10/186 45Ca-uptake(103cpm/mg Pr)Ca content(nmol/mg Pr)ALP activity(U/mg Pr)Control32.25.2046.87.2
7、37411CalcifiedVSMC88.210.21*102.218.23*62484*Table:General index of calcified VSMCs.(n=6,XS.D.)ALP:alkaline phosphatase.*p0.01 vs.control.2020/10/1870510152025controlcalcificationADM content(fmol/mL)*conVSMC0.00.51.01.52.02.5conVSMC*relative amount of ADM mRNA(%)ADM content in calcified VSMCADM mRNA
8、 relative amount in calcified VSMCCRLR mRNARAMP2 mRNARAMP3 mRNA-actin mRNA446 bp416 bp371 bp291 bpC CRLR C RAMP2 C RAMP3CRLRRAMP2RAMP30.00.51.01.52.02.53.03.54.04.5conVSMC*relative amount of X mRNA(%)Alterations of CRLR,RAMP2，RAMP3 mRNA in calcified VSMCs mRNA expression were determined by semi-quan
9、titative RT-PCR analysis.CRLR:calcitonin receptor like receptor.RAMP:receptor activity modifying protein.*p0.01,compared with control.2020/10/188024681012ControlCalcifyingCa-ADM ICa-ADM IICa-ADM IIICa-ADM IVCell counts(104 cells/mL)0123456ControlCalcifyingCa-ADM ICa-ADM IICa-ADM IIICa-ADM IV3H-TdR i
10、ncorporation012345Control Calcifying Ca-ADM I Ca-ADM II Ca-ADM III Ca-ADM IV 3H-Leu incorporation*Exogenous ADM inhibit calcification of VSMC2020/10/1892020/10/1810Cell counts(104cells/ml)3H-TdR(103cpm/mg Pr)3H-Leu(103cpm/mg Pr)45Ca uptake(103cpm/mg Pr)C a l c i u m content(nmol/mg Pr)ALP activity(U
11、/mg Pr)Calcifying 10.02+1.164.86+0.52 3.66+0.40 169.4+13.1 177.4+20.0588+67 Ca-ADM IV 8.12+0.963.32+0.28 3.02+0.26 126.7+11.7 104.5+9.7 327+28Ca-ADM IV-PKAI 9.66+0.88#4.92+0.52#3.42+0.28#110.7+9.9#112.7+10.6317+26#p0.05,#p0.01,compared with the group having the same peptide but no inhibitor.Calcifyi
12、ng:calcifying alone group;Ca-:calcifying;ADM:Adrenomedullin.PKAI:protein kinase A inhibitor.Effects of protein kinase A inhibitor on vascular calcification2020/10/1811von Kossa staining for myocardium and aortaA and C:control;B and D:calcified myocardium and aorta respectivelyABCD2020/10/1812control
13、 Calcification Body Wt,g2871324712*LVW/BW,mg/g2.490.072.70 0.05*Plasma Ca content,mmol/L2.210.10 2.250.08Heart Ca content,mol/g dw 2.160.13 9.761.32*Aortic Ca content,mol/g dw11.991.98 84.7612.09*Myocardial ALP activity,U/mg Pro153.796.96 255.9983.41*Aortic ALP activity,U/mg Pro 180.4334.92329.64105
14、.8*General characteristics of calcification of myocardium and aorta in rats 2020/10/1813controlcalcification MBP,mmHg 1047 1092 LVEDP,mmHg 4.621.60 5.101.23+LVdp/dtmax,mmHg/s 4386304 3977276*HR,beats/min 41417 41415-LVdp/dtmax,mmHg/s 4033387 3227315*Blood pressure and cardiac function of control and
15、 vascular calcification of rat.2020/10/1814051015controlcalcificationPlasma ADMcontent(fmol/mL)*0.80.850.90.9511.051.1controlcalcificationmyocardial ADMcontent(pg/mg pro)*00.20.40.60.811.2controlcalcificationaortic ADMcontent(pg/mg pro)*ADM content in plasma,myocardium and aorta2020/10/1815ADMCRLRRA
16、MP2 RAMP30.00.51.01.52.02.53.03.5controlcalcification*X mRNA/B-actin mRNA amount(%)Relative amount of ADM,CRLR,RAMP2 and RAMP3 mRNA in myocardium and aorta of rats.*p0.05,*p0.01 vs controlADMCRLRRAMP2 RAMP3012controlcalcification*X mRNA/B-actin mRNA amount(%)MyocardiumAorta2020/10/1816Aorta Ca(umol/
17、g dw)Myocardium Ca(umol/g dw)Aorta ALPActivity(U/mg Pro)Control15.2 1.062.41 0.1846 5.7VDN116 8.42*7.66 0.82*627 38*VDN+ADM78.6 5.17#4.84 0.42#371 48#VDN+Ls122 8.827.22 0.68597 44*p0.05,VDN vs control;#p0.05,VDN+ADM vs VDN and VDN+LsVDN:vitamin D3 plus nicotine;Ls:liposome;ADM:adrenomedullinAdrenome
18、dullin inhibits vascular calcification02468ControlVDN VDN+ADM VDN+Ls Aortic ADM(pg/mg Pro)#*0100200300400500600ControlVDN VDN+ADM VDN+Ls Plasma ADM (pg/mL)*p0.05,VDN vs control;#p0.01,VDN+ADM vs VDN and VDN+Ls2020/10/1817FigB:The Scatchard plots from 125I-ADM binding with vascular membrane(-)ADM(+)A
19、DM(10-7mol/L)Control1.04 0.082.32 0.14VDN0.82 0.08*1.64 0.20*VDN+ADM1.58 0.21#2.86 0.38#VDN+Ls0.95 0.061.98 0.28Vascular cAMP content Vmax(fmol/mg)Kd(nmol/L)Control833 671.4 0.2VDN606 52*2.3 0.3*VDN+ADM713 48#1.5 0.2#VDN+Ls522 661.8 0.32020/10/1818Fig.1.Effect of ADM(2752)on calcific nodules in thor
20、acic aorta of rats(n=7/group,all von Kossa sections132).A.None of calcific nodules B.in control rats;B.dispersed calcific nodules in model rats;C.dense calcific nodules in model rats;D.less dispersed calcific nodules in ADM rats;E.lest calcific nodules inADM and ADM(2752)rats;F.less dispersed calcif
21、ic nodules in ADM(2752)rats.2020/10/18192020/10/18202020/10/1821nEndogenous ADM was increased,ADM and its receptor gene expressions were upregulated in calcified VSMC and aortanExogenous ADM can inhibit vascular calcificationnMechanism of ADM inhibiting calcification might be involved in cAMP/PKA pathwasy nFragments derived from ADM can also inhibit vascular calcificationSummary2020/10/1822谢谢您的聆听与观看THANK YOU FOR YOUR GUIDANCE.感谢阅读！为了方便学习和使用，本文档的内容可以在下载后随意修改，调整和打印。欢迎下载！汇报人：XXX日期：20XX年XX月XX日
