Effects of Shenshuaikang Enema Liquid on the Wnt/β-catenin pathway for inhibiting the hypoxia reoxygenation injury in HK-2 cells / 中成药

AIM To investigate the protective effect of serum containing Shenshuaikang Enema Liquid (Rhei Radix et Rhizoma,Salviae miltiorrhizae Radix et Rhizoma,Carthami Flos,Astragali Radix) on HK-2 cells injured by hypoxia/reoxygenation (H/R) and its effect on Wnt/β-catenin pathway.MEHTODS Six New Zealand white rabbits randomly divided into control group,Shenshuaikang high dose group and PBS group (n =2) were treated accordingly for serum collection after 3 days' enema.The HK-2 cells injured by hypoxia/reoxygenation were then administered with serum of rabbits from the control group,PBS group,Shenshuaikang groups (high dose,middle dose and low dose groups due to the differently diluted concentrations) respectively.The H/R damage was determined by ROS fluorescence probe,the cellular damage/apoptosis were analyzed by CFSE/PI method and flow cytometry combined with Annexin V-FITC/PI,and the investigation on effects of drugs on expression of Wnt4 mRNA and β-catenin mRNA were accomplished by fluorescence quantitative PCR.RESULTS The fluorescence intensity of intracellular ROS expression was significantly increased after modeling.CFSE/PI double staining showed that the variant Shenshuaikang dose groups displayed obvious proportional mortality superiority to either the control group or PBS group,and the high dose group achieved the lowest mortality.Annexin V-FITC/PI and flow cytometry showed that,at 12 h,compared with the control group (45.6 ± 2.2)％ and PBS group (41.6 ±0.7) ％,Shenshuaikang groups [low dose group (14.8 ± 0.3) ％,middle dose group (10.3 ± 0.6) ％,high dose group (12.9 ± 0.9)％] obviously inhibited apoptosis/death.Shenshuaikang Enema Liquid medicated serum demonstrated its significant effect on the increase of the Wnt4 mRNA expression and a dual-directional regulation on the expression of β-caterin mRNA by quantitative PCR (P ＜ 0.05).CONCLUSION Inhibition of the apoptosis/death of HK-2 cells with hypoxia/reoxygenation injury due to the serum containing Shenshuaikang Enema Liquid suggests the agent's influence on the activation of Wnt/β-catenin pathway.

Expression of Brain Natriuretic Peptide in Myocardial Tissue of Sudden Cardiac Death and Non-sudden Cardiac Death / 法医学杂志

Objective To study the expression pattern of brain natriuretic peptide (BNP) in myocardial tissue from forensic routine cases and to explore its application value in the forensic determination of sudden cardiac death (SCD). Methods The data of 96 autopsy cases accepted by the center of Medico-legal Investigation of China Medical University between December 2008 to May 2014 were collected. There were 62 cases in SCD group cardiac and 34 cases in non-SCD group. The myocardial tissues were taken from left and right ventricular wall, respectively. The expressions of BNP protein and BNP mRNA in myocardial tissue were detected by HE staining, immunohistochemical staining, Western blot-ting and quantitative real-time reverse transcription-polymerase chain reaction (RT-qPCR), etc. Results The immunohistochemical staining of myocardial tissue showed diffusely positive staining in SCD group, and patchily or diffusely positive staining in non-SCD group with lighter degree. The result of Western blotting showed that the expression of BNP protein elevated in left ventricular wall of SCD group. The result of RT-qPCR showed a positive correlation between the BNP mRNA expressions in bilateral ven-tricular walls and the heart weight, bilateral lung weight, and N-terminal prohormone of brain natriuretic peptide (NT-proBNP) concentration. There were large differences between the BNP mRNA concentra-tions in SCD group and non-SCD group, and the former was statistically higher (P<0.05). Conclusion The expressions of BNP protein and BNP mRNA in myocardial tissue are related to the causes of death. Combined with pathological changes, the expressions of BNP protein and BNP mRNA in myocar-dial tissue have certainly practical significance for the determination of SCD and the analysis of the death mechanism in the cases related to forensic pathology.

The Changes of HIF-1α and VEGF-A in Myocardial Tissue of Rats with Arrhythmias / 法医学杂志

OBJECTIVES@#To observe the expression changes of hypoxia inducible factor-1α（HIF-1α） and vascular endothelial growth factor-A （VEGF-A） in rats with arrhythmias, and to explore the differences of the expression pattern in the two indicators of acute myocardial ischemia caused by arrhythmias and coronary insufficiency.@*METHODS@#The arrhythmia was induced by CaCl₂, and the expression changes of HIF-1α and VEGF-A were detected by immunohistochemistry, Western blotting and real-time PCR within 6 h after the arrhythmia in rats.@*RESULTS@#The expression of HIF-1α and VEGF-A showed diffuse in the myocardial tissue of rats died from arrhythmias. Both of them increased in the early arrhythmia, then decreased. Extensive myocardial ischemia happened at the beginning of arrhythmia occurrence and its range didn't expand with time.@*CONCLUSIONS@#The expressions of HIF-1α and VEGF-A in myocardium of the rats with arrhythmia can provide evidence for the differential diagnosis of acute myocardial ischemia caused by fatal arrhythmia and coronary insufficiency.