| [objective]:chronic hypoxia is one of most important factors that cause ventricular remodeling . ventricular remodeling is important pathological molecular mechanism which lead to most heart disease running to cardiac failure and arrhythmia,It's also the sign of aging heart.BMSCs transplantation is the dependable treatment of ventricular remodeling now.BMSCs can instead of Apoptosis by differentiating to cardiom Yocytes;It can also form new blood vessel to improve Partial blood circulation by differentiating to vascular endothelial cell and secreting much of cytokines; It can improve the local condition by controlling the inflammatory response;It also have the effects of anti-apoptosis.So BMSCs transplantation can improve ventricular remodeling by above mechanism. The PNUTS (Serine/threonine protein phosphatase 1 regulation 10)expression in BMSCs has the effects of anti-apoptosis and recovering cardiac function of post-acute myocardial infarction.But now , there is not any study about BMSCs improving ventricular remodeling caused by chronic hypoxia with PNUTS and the correlation between PNUTS and ventricular remodeling caused by chronic hypoxia. This topic establish SD rat ventricular remodeling caused by chronic hypoxia model by Automatic oxygen device,and the models are treated with BMSCs to establish treatment group. We detect miR-34a's expression? pnuts mRNA's expression? PNUTS concentration in ventricle of ventricular remodeling rats caused by chronic hypoxia , and P I CP? PIIINP concentration in serum and cardiac muscle tissue. We treat ventricular remodeling rats caused by chronic hypoxia with BMSCs,then detect miR-34a's expression?pnuts's expression? PNUTS concentration in ventricle and and P I CP? PIIINP concentration in serum and cardiac muscle tissue in 4 weeks so that we study whether BMSCs improving ventricular remodeling caused by chronic hypoxia with PNUTS and the correlation between PNUTS and ventricular remodeling caused by chronic hypoxia.[Methods]45 SD rats are randomly divided into 9 groups,each group have 5 rats.Group A(control group)?Group B(hypoxia 7 days)?Group C(hypoxia 14 days)?Group D(hypoxia 21 days)? Group E(hypoxia 28 days)? Group B'(intraperitoneal injection BMSCs after hypoxia 7 days by 5×106/mL,1mL,treat 4 weeks)? Group C'(intraperitoneal injection BMSCs after hypoxia 14days by 5×106/mL,1mL,treat 4 weeks)?Group D'(intraperitoneal injection BMSCs after hypoxia 21 days by 5×106/mL,1mL,treat 4 weeks)?Group E'(intraperitoneal injection BMSCs after hypoxia 28 days by 5×106/mL,1mL,treat 4 weeks).We establish SD rat ventricular remodeling model by Automatic oxygen device , treating the rats with BMSCs for 4 weeks ,then we get rats'weight. Inject 10%chloral hydrate of 0.3mL/100g in intraperitoneal and then dissect rats.open the abdomen and get the heart to make pathological section.We observe Cell morphology, edema, hypertrophy, degeneration,necrosis and inflammatory cells infiltration in cardiac muscle tissue with HE staining.We observe Myocardial collagen distribution of every rat's cardiac muscle tissue by masson staining and calculate CVF.Fluorescent Tunel detect the hypoxia group and treatment group rats myocardial apoptosis in myocardial tissue,cell apoptosis rate calculation.Using qPCR detection the hypoxia group,the treatment group rats myocardial tissue and miR-34a express,pnuts mRNA expression in BMSCs.Using ELISA to detect the hypoxia group and treatment group rats serum P I CP, concentration of PIIINP, P I CP in myocardial tissue, concentration of PIIINP,PNUTS concentration, concentration PNUTS in BMSCs.Analysis relationship between miR-34a content,PNUTS mRNA content,PNUTS concentration and the concentration of P I CP, PIIINP, collagen volume fraction and the cell apoptosis rate.[Results]1?Compared with control group,the rats serum and myocardial tissue hypoxia group of P?CP, P?NP increased significantly (P<0.01); Oxygen to the comparative analysis between two groups, according to the P I CP, PIIINP in serum and myocardial tissue significant difference (P<0.01), and a lack of oxygen increases with time.2?Compared with control group, HE staining showed the rat myocardial cell hypoxia group were different degree of edema, vacuoles degeneration, hypertrophy,inflammatory cells infiltration and fibrosis; Masson staining showed collagen increased myocardial cell gap, CVF of hypoxia group significantly incresed (P <0.0 1).pathological section The percentage of TUNEL show Increase myocardial cell apoptosis,myocardial cell apoptosis rate increased significantly (P < 0.01). Among various hypoxia group two comparative analysis shows that: the myocardial pathological changes,the degree of fibrosis,CVF,myocardial apoptosis rate differ significantly (P < 0.01),and as time increases.3?Compared with control group, the myocardial tissue hypoxia group miR-34 a expression quantity increased significantly (P<0.01), pnuts mRNA expression quantity significantly reduced (P<0.01); Between different hypoxia group two comparison shows: miR-34 a expression of significant difference (P<0.01), and lack of oxygen increases with time, pnuts mRNA expression of significant difference(P<0.01), and reduce the oxygen increases with time.4?Correlation analysis showed that rats serum and myocardial tissue P?CP, P?NP concentration and CVF were significantly positive correlation (P<0.01); P?CP,concentration of P?NP and myocardial apoptosis was significantly positively related to cell apoptosis rate (P<0.01).5?Stem cells in the miR - 34 a express the amount was significantly lower than the control group (P<0.01),Stem cells pnuts quantity, concentration of pnuts mRNA expression is significantly higher than control group (P<0.01).6?The stem cell treatment group in serum and myocardial tissue P?CP,concentration of P?NP the hypoxia group significantly decreased (P < 0.01).7?Stem cell treatment group B' group C' group D' group E ' abnormal myocardial tissue pathological change,degree of fibrosis of the corresponding the hypoxia group improved significantly, the myocardial tissue of CVF and myocardial apoptosis rate dropped significantly in the corresponding the hypoxia group(P<0.01).8?Stem cell treatment group B' group C'group D' myocardial tissue miR-34a expression quantity significantly reduced in the corresponding the hypoxia group(P<0.01 ),pnuts mRNA expression quantity increased significantly (P<0.01),pnuts concentration increased significantly (P<0.01); Stem cell therapy group E'myocardial tissue miR-34a express, pnuts mRNA expression and pnuts concentration group E in the hypoxia group no significant change (P > 0.01).9?Correlation analysis showed that: in addition to the group E (hypoxia group 28 days), myocardial tissue hypoxia group and treatment group and miR-34a expression quantity and pnuts mRNA expression quantity has significant negative correlation(P<0.01), pnuts content and hypoxia group and treatment group P?CP, content of P?NP, CVF,cell apoptosis rate was significantly negative correlation(P<0.01).Linear regression analysis: hypoxia group and treatment group miR-34a was associated with a significant content of PNUTS (R2=0.856, R2=0.862), the regression coefficients were 0.132 (P < 0.05); 0.162 (P < 0.162); PNUTS content and oxygen P?CP, P?NP levels significantly correlated (R2=0.986, R2=0.949) regression coefficient respectively 22.518 (P<0.05), 0.05 (P<0.05); PNUTS content and treatment group P?CP, P?NP levels significantly correlated (R2=0.992, R2=0.981)regression coefficient respectively 7.673 (P<0.05), 0.05 (P<0.05).[Conclusion]1?It's successful to establish the ventricular remodeling model caused by chronic hypoxia, In 21 days of oxygen to hypoxia 28 days ventricular reconstruction is most serious.2?Chronic lack of oxygen in the process of the ventricular remodeling, myocardial fibrosis, myocardial apoptosis appeared almost the same time, both to participate in the development of ventricular remodeling process, and the lack of oxygen to 28 days of oxygen increase up to 21 days.3?BMSCs can improve the ventricular remodeling model caused by chronic hypoxia,but it can't recover ventricular remodeling.4?miR-34a can control pnuts expression.5?miR-34a is low expressions in BMSCs,pnuts is high expressions in BMSCs.6?It's obvious correlative between PNUTS and ventricular remodeling model caused by chronic hypoxia.7?BMSCs can improve the ventricular remodeling model caused by chronic hypoxia by pnuts adjusted with miR-34a.But for oxygen chronic hypoxia model for a longer time, BMSCs can not improve the ventricular remodeling model caused by chronic hypoxia by pnuts adjusted with miR-34a. |
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