Abstract:Aim To explore whether 1,5-(OH)2D3 can regulate the autophagy and calcification of human umbilical vein endothelial cells (HUVEC) and the calcification of mouse atherosclerotic plaque through Ca2+/CaM signal. Methods (1) HUVEC were incubated in high fat microenvironment to stimulate autophagy. The autophagy level was detected by Western blot, fluorescence microscopy, transmission electron microscopy after HUVEC being stimulated by 1,5-(OH)2D3. Lentivious were used to overexpress or silence calmodulin (CaM), and the calcification was tested by Western blot,calcium ion detection kit, Alizarin red. (2) ApoE－/－ mice fed with high fat diet were randomly divided into 3 groups:control group (1,5-(OH)2D3 2.5 μg/(kg·d) dissolved in 0.1 mL soybean oil, Gavage)； CaM overexpressed group (control +CaM overexpressed lentivirus, Caudal vein injection) and CaM silenced group (control+CaM silenced lentivirus, Caudal vein injection). Scanning electron microscope was used to observe autophagosomes and calcification of the aorta. HE and Von Kossa staining were used to detect the As degree and calcification of the aorta, respectively. Results The autophagy was promted and the expression of Ca2+ and CaM were up-regulated after HUVEC being stimulated by 1,5-(OH)2D3. Compared with the control group, CaM overexpressed group showed enhanced autophagy and reduced calcification in HUVEC and As plaque in ApoE－/－mice. While the CaM silencing group showed autophagy dysfunction, increased calcium depositions in HUVEC and plaques. Conclusion 1,5-(OH)2D3 regulates HUVEC autophagy to inhibit cell calcification and plaque calcification through Ca2+/CaM signaling.

Abstract:Aim To study the relationship between Ca2＋/calmodulin-dependent protein kinase Ⅳ （CaMKⅣ） in Xinjiang Kazakh populations and patients with essential hypertension （EH）. Methods Based on case-control study,70 patients with EH and 65 subjects with normal blood pressure were selected from Hankazi township of Xinjiang where Kazakh people live relatively fixed. By physical examination and questionnaire,basic data and blood samples were collected. Serum CaMKⅣ level was detected by using double antibody sandwich enzyme linked immunosorbent assay. Results Serum CaMKⅣ level in EH group was significantly lower than that in normal blood pressure group （P<0.01）. There were no correlation between serum CaMKⅣ and total cholesterol,triglyceride,low density lipoprotein cholesterol,high density lipoprotein cholesterol （all P>0.05）. There were no correlation between serum CaMKⅣ and systolic blood pressure （SBP）,diastolic blood pressure （DBP） in the normal blood pressure group. There were negative correlation with SBP,DBP in EH group,more obviously correlated with the latter （r＝－0.291,－0.381,all P<0.05）. CaMKⅣ had more obvious correlation with patients whose diastolic blood pressure was no less than 100 mmHg （r＝－0.411,P<0.05）. Logistic regression analysis showed that higher serum CaMKⅣ had evidently protective effect to essential hypertension （OR was 0.930,95%CI was 0.909～0.952）. Conclusions Serum CaMKⅣ has independently negative correlation with EH of Kazakh populations in Xinjiang,and it is an independent factor of EH.

Abstract:Aim To study the effects of tetramethylpyrazine(TMP) on calmodulin(CaM) and calcinuerin(CaN) in the proliferation of vascular smooth muscle cell(VSMC) induced by angiotensinⅡ(AngⅡ). Methods A cell proliferating model of VSMC induced by angiotensinⅡ(AngⅡ) was established;the varity of CaM and CaN activities affected by tetramethylpyrazine(TMP) at different time and different concentration was observed by enzyme reaction phosphorus measurement. Results Cell proliferation activity,CaM and CaN activities were increased significantly in VSMC proliferation induced by AngⅡ(p<0.01).While treated with TMP,the index were obviously reduced compared with AngⅡ group(p<0.01). Conclusions The VSMC proliferation induced by AngⅡ can be inhibited by TMP significantly,and the inhibiting mechanism of TMP may be related to inhibiting CaM and CaN activities then restraining the proliferation of VSMC in a dose and time-dependent manner.

Abstract:Aim To investigate the effects of platelet-derived growth factor-BB (PDGF-BB)on content of calmodulin in human vascular smooth muscle cells. Methods The human vascular smooth muscle cells were cultured and effects of PDGF-BB on content of calmodulin in human vascular smooth muscle cells were assayed at different time and different concentration by its ability to stimulating calmodulin dependent cyclic nucleotide phsphodiesterase in vitro. Results Calmodulin level of quiescent human vascular smooth muscle cells was increased in its cell cycles with a maximal response 9 h, and PDGF-BB could significantly increase content of calmodulin of human vascular smooth muscle cells in a dose-dependent manner, with a maximal response at a concentration of 30 μg/L. Conclusion PDGF-BB could significantly increase calmodulin level of quiescent human vascular smooth muscle cells in a dose-dependent manner.