Effect of Celastrus orbiculatus extract on proliferation and apoptosis of multiple myeloma cells and its mechanism / 中国药理学通报

Aim To explore the effeet of the extract of Celastrus orbiculatus extract on the proliferation anrl ap- optosis of multiple myeloma eells (U-1996) and its molecular mechanism.Methods The U-1996 eells were divided into normal control ( NC ) group, the southern snake vine extraet group, si-NC group, si- LINC00472 group, pcDNA-NC group, pcDNA- LINC00472 group, southern snake vine extraet + si-NC group,and southern snake vine extraet + si-LINC00472 group.Heal-time quantitative PCR was used to deteet LINC00472 expression, CCK-8 and flow cytometry to deteet eell proliferation and apoptosis, Western blot to deteet the expression levels of phosphorvlated phos- phatidvlinositol-3 -hydroxykinase (p-PI3K) and phos- phorylated protein kinase B ( p-Akt).Results After treated with Celastrus orbieulatus extract, the viability of U-1996 eells and the expression of p-PI3K and p- Akt were significantly reduced, the apoptotie rate and the expression of LINC0047 were significantly raised ( P < 0.05 ).After inhibiting the expression of LINC00472, the viability of U-1996 eells and the ex¬pression of p-PBK and p-Akt significantly inereased, the apoptotie rate and the expression of LINC0047 sig¬nifieantly deereased ( P <0.05 ).After overexpression of LINC00472, the viability of U-1996 eells and the ex¬pression of p-PI3K and p-Akt were signifieantly re- dueed, the apoptotie rate and the expression of LINC0047 significantly inereased ( P <0.05 ).Inhibi¬ting the expression of LINC0047 can reverse the effects of Celastrus orbiculatus extract on proliferation , apopto- sis and PI3K/AKT signaling pathway of U-1996 cells (P < 0.05 ).Conclusions Celastrus orbiculatus ex¬tract can inhibit the proliferation of multiple myeloma cells and induce apoptosis through up-regulating LINC00472 to inhibit PI3K/Akt signaling pathway.

Effect of Zuoguiwan on Expression of Ca2+ Transport Pathway-related Protein in Ovariectomized Rats / 中国实验方剂学杂志

Objective::To explore the effect of Zuoguiwan on the bone mineral density (BMD) and the expressions of Ca2+ transport-associated protein in ovariectomized rats. Method::The 48 female SD rats were randomly divided into six groups: normal group, model group, sham operation group, estrogen group(0.167 mg·kg-1) and low and high-dose Zuoguiwan groups(9.6, 38.4 g·kg-1), with 10 rats in each group. Except for the sham-operated group, the ovariectomized rats in the other groups received the bilateral ovariectomy. Therapeutic intervention was given in each group for 3 months after the establishment of the model. After 12 weeks, BMD was measured using dualenergy X-ray absorptiometry. Tartrated presistant acid phosphatse(TRACP) and serum calcium were detected by biochemical kits.Protein expression in Ca2+ transport (Bone tissue) was detected by Western blot. Result::Compared with the normal group, the serum calcium of the model group was decreased(P<0.01). Compared with the normal group, BMD of the model group was decreased (P<0.01). The serum calcium of rats in high-dose group and western medicine group was higher than that of model group(P<0.01). BMD in model group was lower than that of Zuoguiwan groups and estrogen group(P<0.05). There was no significant difference in TRACP among the groups. Nilestriol and Zuoguiwan can down-regulate the expressions of TRPV5, NCX1, CaBP-D28K and PMCA1b in bone tissue of castrated rats(P<0.05, P<0.01). Conclusion::Zuoguiwan can down-regulate the expressions of Ca2+ transport-associated proteins (Bone tissues) in rat osteoclasts, with an efficacy on osteoporosis.

The effects of low intensity pulsed ultrasound on the expression of calcium transportation associated proteins during dentin-pulp complex injury and repair in rats / 实用口腔医学杂志

Objective: To investigate low intensity pulsed ultrasound (LIPUS) on the expression of L-type calcium ion channels(cav1. 2) and Na +-Ca2 + exchangers(NCX1) during dentin-pulp complex injury and repair in rats. Methods: Cavity preparation was made on the upper right first molar of 40 male adult SD rats,20 of them and the upper left first molar of the other 20 were randomly chosen for LIPUS irradiation(frequency: 1. 5 MHz,200 μs pulses,pulse repetition frequency: 1 KHz,ISATA 30 mW/cm2,20 min /d),so the animals were randomly allocated into 4 groups(n = 10): Control group,LIPUS group,cavity preparation group and cavity preparation + LIPUS group. At 1,3,7,14 d post-irradiation the rats were sacrificed respectively for HE stain and immunohistochemical analysis. Results: Reparative dentin formation was observed at 14 days after cavity preparation and LIPUS irradiation,the expression of Cav1. 2(L-type) and NCX1 in this group were increased significantly at day 1 and day 3. Compared with the control group, the expression of Cav1. 2 in LIPUS group increased at day 1 post-irradiation. Conclusion: LIPUS may enhance tertiary dentin formation and up-regulate the expression of Cav1. 2 and NCX1 at the early period of dentin injury.

Expression of Wnt and NCX1 and its correlation with cardiomyocyte apoptosis in mouse with myocardial hypertrophy

Objective: To study the correlation between expression of Wnt and NCX1 and cardiomyocyte apoptosis in mouse with myocardial hypertrophy. Methods: C57B/16 male mice were given the subcutaneous injection of 1 mg/kg isoprenaline to build the myocardial hypertrophy model. After 14 d of model building, mice were executed by cervical vertebra luxation. The ratio of heart weight/body weight (HW/BW) and heart weight/tibia length (HW/TL) was observed and proved using HE staining that detected the size of cardiomyocytes. 40 male C57B/16 mice were randomly divided into the sham group (normal saline) and model group (isoprenaline), with 20 mice in each group. The terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling was applied to detect the cardiomyocyte apoptosis; while Western blot and immunohistochemistry were employed to detect the expression of Wnt and NCX1. Meanwhile, the correlation between these two proteins and cardiomyocyte apoptosis was explored. Results: Compared with the sham group, the ratio of HW/BW and HW/TL was increased in the model group, as well as the bigger and hypertrophied cardiomyocytes, decreased number and increased apoptosis of cardiomyocytes, and increased positive expression of Wnt3a, Wnt5a and NCX1 in the cardiac muscle tissue. Besides, there was positive correlation between the expression of Wnt and NCX1 and the cardiomyocyte apoptosis. Conclusion: The expression of Wnt3a, Wnt5a and NCX1 in mouse with myocardial hypertrophy is increased and positively correlated with the cardiomyocyte apoptosis.

Expression of Wnt and NCX1 and its correlation with cardiomyocyte apoptosis in mouse with myocardial hypertrophy

OBJECTIVE@#To study the correlation between expression of Wnt and NCX1 and cardiomyocyte apoptosis in mouse with myocardial hypertrophy.@*METHODS@#C57B/16 male mice were given the subcutaneous injection of 1 mg/kg isoprenaline to build the myocardial hypertrophy model. After 14 d of model building, mice were executed by cervical vertebra luxation. The ratio of heart weight/body weight (HW/BW) and heart weight/tibia length (HW/TL) was observed and proved using HE staining that detected the size of cardiomyocytes. 40 male C57B/16 mice were randomly divided into the sham group (normal saline) and model group (isoprenaline), with 20 mice in each group. The terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling was applied to detect the cardiomyocyte apoptosis; while Western blot and immunohistochemistry were employed to detect the expression of Wnt and NCX1. Meanwhile, the correlation between these two proteins and cardiomyocyte apoptosis was explored.@*RESULTS@#Compared with the sham group, the ratio of HW/BW and HW/TL was increased in the model group, as well as the bigger and hypertrophied cardiomyocytes, decreased number and increased apoptosis of cardiomyocytes, and increased positive expression of Wnt3a, Wnt5a and NCX1 in the cardiac muscle tissue. Besides, there was positive correlation between the expression of Wnt and NCX1 and the cardiomyocyte apoptosis.@*CONCLUSION@#The expression of Wnt3a, Wnt5a and NCX1 in mouse with myocardial hypertrophy is increased and positively correlated with the cardiomyocyte apoptosis.

Combined actions of Na+/K+-ATPase, NCX1 and glutamate dependent NMDA receptors in ischemic rat brain penumbra / 대한해부학회지

Instrumental role of Na+ and Ca2+ influx via Na+/K+ adenosine triphosphatase (Na+/K+-ATPase) and Na+/Ca2+ exchanger 1 (NCX1) is examined in the N-Methyl-D-aspartate (NMDA) receptor-mediated pathogenesis of penumbra after focal cerebral ischemia. An experimental model of 3, 6, and 24 h focal cerebral ischemia by permanent occlusion of middle cerebral artery was developed in rats. The changes in protein expression of Na+/K+-ATPase and NCX1 as well as functional subunits of NMDA receptor 2A and 2B (NR2A and NR2B) in the penumbra were assessed using by quantitative immunoblottings. The most prominent changes of Na+/K+-ATPase (78+/-6%, n=4, *P<0.05) and NCX1 (144+/-2%, n=4, *P<0.05) in the penumbra were developed 24 h after focal cerebral ischemia. The expression of NR2A in the penumbra was significantly increased (153+/-9%, n=4, *P<0.05) whereas the expression of NR2B was significantly decreased (37+/-2%, n=4, *P<0.05) as compared with sham-operated controls 3 h after focal cerebral ischemia. However, the expression of NR2A and NR2B in the penumbra was reversed 24 h after focal cerebral ischemia (NR2A: 40+/-7%; NR2B: 120+/-16%, n=4, *P<0.05). Moreover, the decreased expression of neuronal nuclei (NeuN) in the penumbra was most prominent than that of glial fibrillary acidic protein (GFAP) 24 h after focal cerebral ischemia. These findings imply that intracellular Na+ accumulation via decreased Na+/K+-ATPase exacerbate the Ca2+ overload cooperated by the increased NCX1 and NR2B-containing NMDA receptor which may play an important role in the pathogenesis of the penumbra.

The Expression and Implication ofTRPV5, Calbindin-D28k and NCX1 in Idiopathic Hypercalciuria / 华中科技大学学报（医学）（英德文版）

Summary: The expression of calcium epithelium TRPV5, alcium binding protein Calbindin-D28k and Na+/Ca2+ exchanger NCX1 was detected in renal distal convoluted tubule, and their effects on urine calcium reabsorption and the possible pathogenic mechanism in idiopathic hypercalciuria (IH) were investigated. Genetic hypercalciuric stone-forming (GHS) rats were chosen as animal models to study urine calcium reabsorption and IH. The cognate female and male rats that had maximal urine calcium were matched to breed next generation. Twelve GHS rats and 12 normal control (NC) SD rats were selected. Western blot and real time quantitative PCR were used to detect the protein and gene expression of TRPV5, Calbindin-D28k and NCX1 respectively. The expression levels of TRPV5 protein and mRNA in GHS rats were significantly lower than in NC rats (P<0.05). Western blot revealed that the expression levels of Caibindin-D28k in GHS rats and NC rats were 0.49±0.02 and 0.20±0.01 respectively, with the difference being significant between them (P<0.05). By using real time quantitative PCR, it was found that there was no significant difference in Calbindin-28k mRNA expression levels between GHS rats and NC rats (P0.05). There was no significant difference in the NCX1 expression between GHS rats and NC rats (P0.05). It was suggested that TRPV5 and Caibindin-D28k might play an important role in urine calcium reabsorption and IH, but they differently contributed to the pathogenesis: The down-regulation of TRPV5 decreases urine calcium reabsorption, directly leading to loss of the urine calcium and resulting in hypercalciuria, and the increased Calbindin-D28k expression could relieve, neutralize and decrease intracellular Ca2+ concentration to maintain calcium balance. NCX1 is not the key protein in urine calcium reabsorption.

NCX and SERCA2a expression in myocardial tissues in rats with pressure overload-induced left ventricular hypertrophy / 基础医学与临床

Objective To observe the abnormity of heart function in rats with pressure overload-induced left ventricular hypertrophy and the changes of NCX,SERCA2a expression in myocardial tissues. Methods Cardiac hypertrophy was induced by clipping the abdominal aorta in rats. The cardiac hypertrophy was evaluated by Left ventricular weight index(LVWI,left ventricular weight/body weight). NCX, SERCA2a mRNA and protein expressions in left ventricular tissues were determined by half-quantitative RT-PCR and Western blot normalized to abundance of GAPDH mRNA and protein,respectively. Results LVSP and LVEDP were obviously enhanced(P

Expression of NCX and NCKX Isoforms mRNAs in Rat Pituitary Gland / 대한해부학회지

Na(+)-Ca(2+) exchanger plays a fundamental role in controlling the changes in intracellular concentration of Na(+) and Ca(2+) ions. Two different families of Na(+)-Ca(2+) exchanger, NCX (K(+)-independent Na(+)-Ca(2+) exchanger) and NCKX (K+-dependent Na(+)-Ca(2+) exchanger), are known and each family includes several isoforms. But little is known about their expression in pituitary gland. In this study, in situ hybridization with digoxigeninlabeled riboprobe and double-labeled experiments with immunohistochemistry were applied to investigate the expression of NCX and NCKX mRNAs and their distribution in normal rat pituitary gland. NCX2 mRNA hybridization signals were expressed in pars distalis, while both NCX2 and NCKX2 mRNAs expression were strongly observed in pars nervosa. NCX2 and NCKX2 mRNA were also expressed in supraoptic nucleus of hypothalamus. In pars distalis, 68.2% of growth hormone secreting cells was colocalized with NCX2 mRNA, whereas NCX2 mRNA was not found in S100 positive folliculostellate cells. These results suggest that NCX2 in pars distalis and NCX2 and NCKX2 in pars nervosa appear to be involved in endocrine function of pituitary gland.

Studies on anti-early hepatic fibrosis of no donating-oleanlic acid derivatives in rats / 中国临床药理学与治疗学

AIM: To investigate protect effect of the NO donating-oleanlic acid derivatives on hepatic fibrosis in rats. METHODS: The rat model with early hepatic fibrosis was induced by carbon tetrachloride (CCl_4). ZCⅡ_2 at the dose of 128 and 64 mg?kg -1 had been given orally for 30 days. Serum level of the total protein (TP), albumin (ALB), the albumin/globulin (A/G) and ALT, AST. The hyaluronic acid (HA), laminin (LN), the procollagen type Ⅲ (PCⅢ) and the level of MDA, GSH-Px in liver tissues were determined. Pathological examination to reveal the extent of liver damages was observed. [WTHZ]RESULTS: ZCⅡ_2 at the dose of 128 mg?kg -1 increased the serum TP, ALB, and A/G and decreased HA, LN, PCⅢ, ALT, and AST more significantly than the model group, and hepatic pathological injury was abated to some degree. CONCLUSIONS: ZCⅡ_2 at the dose of 128 mg?kg -1 can attenuate liver damages, protect against lipoperoxidation and attenuate hepatic fibrosis induced by CCl_4.

Positive inotropic action of dimethylamiloride in normal rat isolated hearts and papillary muscle and its primary mechanism / 中国药理学通报

Aim To study the inotropic action of DMA in normal rat isolated hearts and papillary muscle and search for its primary mechanism.Methods With Lansendorff-perfusion and isolated papillary muscle perfusion,cardiac performance and the systolic function of papillary muscle were measured to estimate the inotropic action of DMA;L-calcium channel blocker and sodium calcium exchanger(NCX)inhibitor were used to search for its primary mechanism.Results ①(1~20)?mol?L-1 DMA exerted a positive inotropic action in normal rat isolated hearts,that is in Langendorff-perfused hearts,DMA enhanced cardiac performance,i.e.LVSP-LVDP,+dp/dtmax,-dp/dtmax significantly(P

The effects and mechanisms of dimethylamiloride on cardiac function in cardiomegaly rat isolated hearts / 中国药理学通报

Aim To investigate the effects and mechanisms of DMA on cardiac function in cardiomegaly rat isolated hearts.Methods With Langendorff perfusion,the changes in the indicators reflecting cardiac function were observed in cardiomegaly rat isolated hearts.Its probable mechanism was explored with calcium channel blocker and sodium calcium exchange(NCX) blockers.Results DMA(0.5～2 ?mol?L~(-1))enchanced cardiac function and exerted positive effects on LVSP-LVDP,+dp/dt_(max)and-dp/dt_(max) in Langendorff-perfused cardiomegaly rat isolated hearts.The effects of DMA on cardiac function in cardiomegaly rat isolated hearts weren′t blocked by nicardipine-one of calcium channel blockers and were blocked by NiCl_2——one of sodium calcium exchange(NCX) blockers.Conclusion DMA can enhance systolic and diastolic function in cardiomegaly rat isolated hearts.By agitating NCX,DMA enhances cardiac function in cardiomegaly rat isolated hearts,independent of L-calcium channel.