肝细胞生长因子对长时间保存大鼠供体心bcl-2mRNA表达及细胞凋亡的影响

Objective To observe the effect of hepatocyte growth factor on isolated rat heart preservation. Methods Forty SD mrs were randomly divided into two groups : control group (n = 20) with isolated rat heart preserved in 4 ℃ HTK solution and experimental group(n =20) with isolated rat hearts preserved in 4 ℃ HTK + rh-HGF(100 μg/L) solution. Cell apoptosis was detected by TUNEL staining and the expression of bcl-2mRNA was examined by RT-PCR. Results Experimental group showed a lower rate of apoptosis eardiomyocytes (P < 0.01) after reperfusion. Immunohistochemieal analysis revealed that c-met receptor expression was stronger in the HGF-treated myocardium than that in the non-HGF-treated myocardium after storage and was associated with a stronger ex-pression of bcl-2 mRNA. Conclusion The administration of rh-HGF before storage improved cardiac function after prolonged myocardial preservation by preventing apoptosis and enhancing expression of bcl-2 mRNA.     

肝细胞生长因子对长时间保存大鼠供体心bcl-2mRNA表达及细胞凋亡的影响

Objective To observe the effect of hepatocyte growth factor on isolated rat heart preservation. Methods Forty SD mrs were randomly divided into two groups : control group (n = 20) with isolated rat heart preserved in 4 ℃ HTK solution and experimental group(n =20) with isolated rat hearts preserved in 4 ℃ HTK + rh-HGF(100 μg/L) solution. Cell apoptosis was detected by TUNEL staining and the expression of bcl-2mRNA was examined by RT-PCR. Results Experimental group showed a lower rate of apoptosis eardiomyocytes (P < 0.01) after reperfusion. Immunohistochemieal analysis revealed that c-met receptor expression was stronger in the HGF-treated myocardium than that in the non-HGF-treated myocardium after storage and was associated with a stronger ex-pression of bcl-2 mRNA. Conclusion The administration of rh-HGF before storage improved cardiac function after prolonged myocardial preservation by preventing apoptosis and enhancing expression of bcl-2 mRNA.     

肝细胞生长因子对长时间保存大鼠供体心bcl-2mRNA表达及细胞凋亡的影响

Objective To observe the effect of hepatocyte growth factor on isolated rat heart preservation. Methods Forty SD mrs were randomly divided into two groups : control group (n = 20) with isolated rat heart preserved in 4 ℃ HTK solution and experimental group(n =20) with isolated rat hearts preserved in 4 ℃ HTK + rh-HGF(100 μg/L) solution. Cell apoptosis was detected by TUNEL staining and the expression of bcl-2mRNA was examined by RT-PCR. Results Experimental group showed a lower rate of apoptosis eardiomyocytes (P < 0.01) after reperfusion. Immunohistochemieal analysis revealed that c-met receptor expression was stronger in the HGF-treated myocardium than that in the non-HGF-treated myocardium after storage and was associated with a stronger ex-pression of bcl-2 mRNA. Conclusion The administration of rh-HGF before storage improved cardiac function after prolonged myocardial preservation by preventing apoptosis and enhancing expression of bcl-2 mRNA.     

Effect of lipopolysaccharide on expression and characterization of cholecystokinin receptors in rat pulmonary interstitial macrophages

AIM: To investigate the effect of lipopolysaccharide (LPS) on the expression and the binding characteristics of cholecystokinin receptors (CCK-R) in rat pulmonary interstitial macrophages (PIMs). METHODS: The PIMs isolated from rat lung tissues were purified by the collagenase digestion method combined with alveolar lavage and pulmonary vessel perfusion. The expression of CCK-R mRNA was detected by RT-PCR and Southern blot analysis and the binding experiments were performed by radioligand binding assay (RBA). RESULTS: CCK-A receptor (CCK-AR) and CCK-B receptor (CCK-BR) mRNA were detected in rat PIMs and their RT-PCR amplified products had a size of approximately 1.37 kb and 480 bp, respectively. The relative expression of CCK-BR mRNA was higher than that of CCK-AR mRNA after incubation with LPS for 0.5, 2, and 6 h. The expression of CCK-R mRNA could be upregulated obviously by LPS. Southern blot analysis of RT-PCR amplified CCK-AR and CCK-BR mRNA products using [γ-32P]ATP 5'-end-labelled probe showed sp     

Targeting at SUR2B/Kir6.1 subtype of ATP-sensitive potassium channel opener natakalim improves pressure overload-induced heart failure

Objective To explore the new stratigies targeting at SUR2B/Kir6.1 subtype against pressure overload-induced heart failure.Methods Pressure overload-induced heart failure was induced in Wistar rat by abdominal aortic banding(AAB).The effects of natakalim(1,3,9 mg·kg-1·d-1,10 weeks)were assessed on myocardial hypertrophy and heart failure,cardiac histology,vasoactive compounds,and gene expression.Isolated working heart and isolated tail artery helical strips were used to examine the influence of natakalim on heart and resistant vessels.Results Ten weeks after the onset of pressure overload,natakalim therapy potently inhibited cardiac hypertrophy and prevented heart failure.Natakalim inhibited the changes of left ventricular haemodynamic parameters,reversed the increase of heart mass index,left ventricular weight index and lung weight index remarkably.Histological examination demonstrated that there were no significant hypertrophy and fibrosis in hearts of pressure overload rat treated with natakalim.Ultrastructural examination of heart revealed well-organized myofibrils with mitochondria grouped along the periphery of longitudinally oriented fibers in natakalim group rats.The content of serum NO and plasma PGI2 was increased,while that of plasma ET-1 and cardiac tissue hydroxyproline,ANP and BNP mRNA was down-regulated in natakalim-treated rats.Natakalim at concentrations ranging from 0.01-100 μM had no effects on isolated working heart derived from Wistar rats;however,natakalim had endothelium-dependent vasodilation effects on the isolated tail artery helical strips precontracted with NE.Conclusions These results indicate that natakalim improves heart failure due to pressure overload by activating KATP channel SUR2B/Kir6.1 subtype and reversing endothelial dysfunction.     

Effects of rat urotensin Ⅱ on coronary flow and myocardial eNOS protein expression in isolated rat heart

AIM: To examine the effects of urotensin Ⅱ, a recently discovered endogenous peptide, on coronary flow (CF), cardiac function, and endothelial nitric oxide synthase (eNOS) expression in isolated rat hearts. METHODS: Heart was isolated and perfused retrogradely via the aorta in Langendorff mode. Rat urotensin Ⅱ was administered in the perfusion solution. The eNOS content in myocardium was determined by Western blot. RESULTS: Rat urotensin Ⅱ had no effect on the heart rate, left ventricular systolic pressure, left ventricular end-diastolic pressure, or ±dp/dtmax. While rat urotensin Ⅱ dose-dependently increased CF. CF was increased by 11.43 %, 6.67 %, 6.62 %, 6.56 %, 6.36 %, and 5.86 % respectively in a time-dependent manner at 5, 10, 15, 20, 25, and 30 min after injection of rat urotensin II 6.66×10-2μg. The maximal effect on CF was found at 5 min following urotensin II administration. NG-nitro-L-arginine methyl ester (L-NAME) did not prevent the increased CF in response to urotensin II. Rat uroten     

Effect of γ-ray on metabolic enzyme CYP3A1 in rat liver on multiple levels北大核心CSCD

Aim To explore the effect of γ-ray on the mRNA,protein expression levels and metabolic activity level of the key drug metabolic enzyme CYP3A1 in rat liver. Methods Wistar rats were randomly divided into control group, 24 h post-radiation group and 72 h post-radiation group. The experimental group was exposed to total body irradiation of single 6 Gy γ-ray. Blood was collected from the orbital venous plexus for blood routine examination and biochemical analysis 24 h and 72 h after irradiation, and liver tissue was prepared for quantifying expression of CYP3A1 mRNA and liver-specific microRNA (miR-122-5p) through RT-PCR. The expression level of CYP3A1 protein was analyzed by Western blot, and the metabolic activity level of CYP3A1 detected by the specific substrate midazolam combined with LC-MS method. Results Com¬pared with the control group, the weights of the rats in the radiation group significantly decreased, and the number of white blood cells was markedly reduced. Simultaneously, the activities of alanine aminotrans-ferase and alkaline phosphatase continuously descended, as well as the levels of total bilirubin and bile acid significantly increased, which indicated that the liver may be damaged after radiation. The relative expression of CYP3A1 mRNA continued to increase significantly 24 h and 72 h after irradiation. CYP3A1 protein expression and metabolic activity levels showed an obvious increasing trend 24 h after irradiation, and rose significantly 72 h after irradiation compared with the control group. At the same time, the expression of miR-122-5p in liver of rats in the 24 h and 72 h post-radiation group continued to decrease rapidly compared with the control group. Conclusions γ-ray radiation may arouse damage effect on liver, which leads to the continuous up-regulation of the mRNA and protein expression levels of the capital metabolic enzyme CYP3A1 in liver tissue, as well as the elevation of the metabolic activity level. The regulatory mechanism might be related to miR-122-5p. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

Effects of dl-praeruptorin A on nucleus factor-κB and tumor necrosis factor-α expression in ischemia-reperfusion hearts

AIM: To study the effects of dl-praeruptorin A (Pd-Ia) on nucleus factor-κB (NF-κB) activativity and tumor necrosis factor-α(TNF-α)expression in ischemia-reperfusion (I/R) myocardium. METHODS: Langendorff's isolated rat heart was subjected to a 10-min ischemia followed by a 30-min reperfusion. NF-κB activity in nucleus was analyzed by Sandwich Enzyme-Linked Immunosorbent Assay (ELISA). TNF-α level in cytoplasm was measured by     

Daidzein affects proliferation and apoptosis in non-small cell lung cancer cells:role of p53 signaling pathway北大核心CSCD

Aim To investigate the effects of daidzein（DD） on the proliferation and apoptosis of non-small cell lung cancer cells,with a focus on the possible role of the p53 signaling pathway in this regard. Methods CCK-8 method and flow cytometry were used to detect the effects of soy isoflavone crude extract and DD on the viability and apoptosis of HELF and H1299 cells. Gene microarray was used to detect the changes in gene expression after treatment of H1299 cells with DD. GSEA and differential analysis were used to screen the major pathways and key genes. RT-qPCR and Western blot were performed to verify the differences in mRNA and protein expression of key genes（p53 and CASP9） in the major pathways. After p53 inhibitor Pifithrin-α inhibited the expression of p53,the effect of DD on p53 mRNA and protein expression levels was examined,and the proliferative effect on H1299 cells was observed. Results Soy isoflavone crude extract and DD promoted proliferation and inhibited apoptosis of normal lung cells and inhibited proliferation and promoted apoptosis of lung cancer cells. p53 signaling pathway was significantly enriched in the DD-treated group（NES=1.78,P=0.000）,and the expressions of p53 and CASP9 genes were found to be significantly up-regulated in the treated group. Compared with the control group,mRNA expression of CASP9 and p53 significantly increased in both HELF and H1299 cells treated with DD（P<0.05）,and p53 protein expression also increased in HELF cells（P<0.05）. After inhibition of p53 expression,DD significantly increased the mRNA expression of p53 in H1299 and HELF cells（P<0.05） and also markedly increased the expression of p53 protein in H1299 cells（P<0.05）,and it was observed that DD inhibited the proliferation of lung cancer cells. Conclusions DD inhibits the proliferation and promotes the apoptosis of lung cancer H1299 cells,and the mechanism mainly involves the p53 signaling pathway. © 2023 Publication Centre of Anhui Medical University. All rights reserved.