| Heart failure is a disease which seriously threats to human health, in addition to the damage of function and structure of the heart itself, also merges damage of other organs’ functional in body, especially the ischemia of kidney.Ramipril(Ramipril, RAM) is belong to ACEI drugs that expands blood vessel, trimetazidine(TMZ trimetazidine,) can improve myocardial ischemia by influencing the energy metabolism of myocardial.RAM and TMZ is still further researched in renal’s protection.This experiment is to examine the levels of expression of Cyt-C, Bcl-2 and Bax, compared with ramipril,trimetazidine,separated and combined on apoptosis of renal cells in rats with chronic heart failure.Objective:The model of heart failure rat in this experiment was established by abdominal aortic constriction. Then, the pathological changes of rat kidney tissue was observed by hematoxylin-eosin staining,AI was evaluated by TUNEL assay,the expression of Cyt-C protein, Bcl- 2 protein and Bax protein in the renal cells of rats were detected by SP staining and Western method,the expression’s level of all m RNA were explored by RT-PCR.To research the difference of ramipril and trimetazidine alone and combined use on apoptosis of kidney cells in rats with chronic heart failure.Methods:1 we randomly selected 10 rats in 50 clean male Wistar rats as control group, the rest of the rats were chose to implement surgery. The standard of model’s success was that left ventricular end-diastolic pressure was greater than or equal to 15 mm Hg.At the same time, for the blank control group rats were isolated from abdominal aorta ligation. After building model, rats were randomly divided into four group,they respectively are model,ramipril, trimetazidine and combination.2 When the rats of each group were observed to the end,using more conductive recor to measure LVEDP of each rat and accurately record the result in detail.Then,taking out the kidney from abdomen and weighing.3 Using HE staining to observe pathological chang of kidney tissue in various subjects.4 By using TUNEL method to detect apoptosis of renal cells and apoptosis index.5 Detecting the protein expression of Cyt-C protein,Bcl-2 protein and Bax protein in kidney tissue of each rat by the SP and Western method.6 The expression level of Cyt-C m RNA, Bcl-2 m RNA and Bax m RNA were evaluate by RT-PCR in rats.Results:1 The results of LVEDP in each group: the LVEDP of blank control group, model group, ramipril group, trimetazidine group and combination group respectively were:( 5.63±0.98) mm Hg,( 19.62±1.91) mm Hg,(12.24±1.43)mm Hg,(12.03±1.23)mm Hg,(7.48±1.41)mm Hg.The LVEDP of treatment group and model group was increased compared with blank control group(P<0.01); Compared with the model group, the LVEDP in treatment groups decreased obviously(P<0.01);the combination group had the greatest reduction(P<0.01); ramipril group and trimetazidine group did not have difference of statistical(P<0.05).2 The pathological morphological changes in Each groups’ rats was observed by HE staining: the glomerulus and renal tubule cells in blank control group were normal, having neat, tight, clear structure; but renal cells of model group rats were swelling, fall off, loosely arranged, having a disorder structure; Compared with the model group, the treatment group rats were obvious improved and the joint treatment group had the most significantly improve.3 The results of renal cells apoptosis index showed that: five groups’ results respectively were(2.62±0.84)%,(40.82±1.31)%,(20.02±1.14)%,(20.10±1.20)% and(14.27±1.40)%. Compared with blank control group, the AI in model and other treatment groups were higher significantly(P<0.01); Compared with model group, the AI of treatment groups were lower significantly(P<0.01);The most obvious reduction of AI was combination group(P < 0.01).4 The expression level of Bcl-2 protein and Bax protein in renal tubule cells in all groups:the trend of Bcl-2 protein expression level in this five groups was:the sham group showed strongly positive, having the most obviously tan particle deposition in microscope; However, the model group had no obvious positive reaction, the treatment groups had visible positive reaction, but the positive intensity weaker than the control group, thus, the combination group with a strong positive reaction in the single drug groups. The results of Bax protein expression showed that the renal tubule cells in control group had no obvious tan particlesd;The cells in model group had obvious positive reaction;Other drug groups only had weak tan particle deposition, at the same time, the combination group was the weakest.5 Cyt-C protein expression was detected by Western method: the results in blank control, model, ramipril, trimetazidine and combination group respectively were 0.32±0.12,2.53±0.10,1.50±0.11,1.58±0.12,0.75±0.06, compared with blank control group, the Cyt-C protein expression levels in after four groups were significantly increased(P<0.01),compared with model group,the medicine groups were significantly lower(P<0.01),and the combination group was the most remarkablely decreasing.6 The Cyt-C,Bcl-2 and Bax m RNA expression were observed by RT-PCR in kidney cells: Cyt-C, Bax m RNA expression trend were similar, compared with the sham group, the expression level of both in model group were increased obviously(P<0.01),but the m RNA level in the treatment groups compared with model group significantly decreased(P<0.01), Especially, the combination group were lower than other drug intervention groups. Compared the sham group, the Bcl-2 m RNA level of the rest groups decreased sinificantly(P<0.01),and the Bcl-2 m RNA expression level in the model group was the lowest(P<0.01), the Bcl-2 m RNA expression level of combination group was highest in treatment groups,the three indicators in the ramipril and trimetazidine groups had no significant difference(P>0.05).Results:1 The occurrence of apoptosis in renal cells is closely related to the damage of renal function in chronic heart failure rats.2 Ramipril and trimetazidine can protect renal function through inhibiting apoptosis,and the effect of combination is better than the utility of medication respectively,hence,they have synergy which is proposed for the new treatments of impaired renal function in chronic heart failure. |
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