肾上腺髓质素对血管钙化的抑制作用优选课件

1、Adrenomedullin inhibits vascular calcification Department of Physiology and Pathophysiology, Peking University Health Science CenterKey Laboratory of the Ministry of Education on Molecular CardiologyYong-Fen Qi2020/10/181Adrenomedullin inhibits vascuVascular calcificationHypertensionDiabetes Mellitu

2、s HyperlipemiaRenal Failure Aging MediaIntima Vascular calcification is believed to be an important risk factor of cardiovasculardiseasesAtherosclerosis2020/10/182VascularHypertensionDiabetes Vasoactive peptides:Adrenomedullin?2020/10/183Vasoactive peptides:?2020/10/1Adrenomedullin (ADM) is a peptid

3、e capable of inducing a potent and long lasting hypotensive effect ADM is mainly synthesized and secreted from vascular endothelial and smooth muscle cells (VSMC) ADM has been shown to have vasodilatory, hypotensive and growth regulating properties, Exogenous ADM or ADM gene delivery attenuated hype

4、rtension and protected against cardiovascular remodeling, but ADM gene knockout resulted in higher blood pressure and cardiovascular abnormalities. Endogenous ADM is an important factor in regulating cardiovascular and renal homoestasis, and is a potent cardio-reno-protective factor2020/10/184Adreno

5、medullin (ADM) is a peptreceptors bound with ADM were orphan L-1 and RDC-1 receptors In addition, another orphan receptor 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 2020/10/185r

6、eceptors bound with ADM were Fig. 1. (A) Calcification of multicellular nodules. von Kossa staining for calcification, showing positive staining as black/brown areas within the main, large, nodular structure (original magnification 40). (B) Formation of multicellular nodules in culture phase contras

7、t microscopy showing rat VSMCs (original magnification 20).AB2020/10/186Fig. 1. (A) Calcification of m45Ca-uptake(103cpm/mg Pr)Ca content (nmol/mg Pr)ALP activity(U/mg Pr)Control32.25.2046.87.237411CalcifiedVSMC88.210.21*102.218.23*62484*Table :General index of calcified VSMCs.( n=6, XS.D.) ALP: alk

8、aline phosphatase. *p0.01 vs.control. 2020/10/18745Ca-uptakeCa content ALP act* ADM content in calcified VSMCADM mRNA relative amount in calcified VSMCCRLR mRNARAMP2 mRNARAMP3 mRNA-actin mRNA446 bp416 bp371 bp291 bpC CRLR C RAMP2 C RAMP3 Alterations of CRLR, RAMP2，RAMP3 mRNA in calcified VSMCs mRNA

9、expression were determined by semi-quantitative RT-PCR analysis. CRLR: calcitonin receptor like receptor. RAMP: receptor activity modifying protein. *p0.01,compared with control.2020/10/188* ADM content in ADM mRNA re*Exogenous ADM inhibit calcification of VSMC2020/10/189*Exogenous2020/10/18102020/1

10、0/1810Cell counts (104cells/ml) 3H-TdR(103cpm/mg Pr) 3H-Leu(103cpm/mg Pr) 45Ca uptake(103cpm/mg Pr) Calcium content(nmol/mg Pr) ALP activity(U/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

11、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. Calcifying: calcifying alone group; Ca-: calcifying; ADM: Adrenomedullin.PKAI: protein kinase A inhibitor.Effects of protein kinase A inhibitor on vascular calc

12、ification2020/10/1811Cell counts (104cells/ml) 3H-Tvon Kossa staining for myocardium and aortaA and C: control; B and D: calcified myocardium and aorta respectivelyABCD2020/10/1812von Kossa staining for myocardcontrol Calcification Body Wt, g2871324712*LVW/BW, mg/g2.490.072.70 0.05*Plasma Ca content

13、, 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 180.4334.92329.64105.8* General characteristics of calcification of myocardium and aorta in

14、rats 2020/10/1813control Calcification Body Wt,controlcalcification 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 vascular calcification of rat. 2

15、020/10/1814controlcalcification MBP, mmHg*ADM content in plasma, myocardium and aorta2020/10/1815*ADM content in plasma, myRelative amount of ADM, CRLR, RAMP2 and RAMP3 mRNA in myocardium and aorta of rats. *p0.05,*p0.01 vs controlMyocardiumAorta2020/10/1816Relative amount of ADM, CRLR, Aorta Ca(umo

16、l/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: adrenomedull

17、inAdrenomedullin inhibits vascular calcification#*p0.05, VDN vs control; # p0.01, VDN+ADM vs VDN and VDN+Ls2020/10/1817Aorta Ca(umol/g dw)MyocardiumFigB: The Scatchard plots from 125I-ADM binding with vascular membrane(-)ADM(+)ADM(10-7mol/L)Control1.04 0.082.32 0.14VDN0.82 0.08*1.64 0.20*VDN+ADM1.58

18、 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 0.32020/10/1818FigB: The Scatchard plots fromFig. 1. Effect of ADM(2752) on calcific nodules in thoracic aorta of rats (n =7/ group, all von Kossa sections132). None of calcific nodules 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.