| Chronic congestive heart failure was the final stage of various heart disease. The morbility and mortality of CHF increased year by year accopanied by the aging of the world population. Although there were many drugs and treatments which could relieve the symptoms of CHF, but drugs or treatments which could really block or reverse the course of CHF were still in need. Therefore study on the pathogenesis and therapeusis of CHF will go on.Statins were hypolipemia drugs applied widely in clinical practice. Some recent studies have indicated statin may be benificial to CHF patients. The present study investigated the effects of aldosterone on proliferation, DNA synthsis, cell cycle change and cllagen synthsis in cultured neonatal rat cardiac fibroblasts with chromatometry, flow cytometry, Western blot, and cell immunofluorescence technique, and the potential molecular mechanism. Meanwhile, the current study investigated the effects of HMG-CoA reducase inhibitor atorvastatin on proliferation, DNA synthesis, cell cycle change, and collagen synthsis in cultured neonatal cardiac fibroblasts induced by aldosterone, and the potential molecular mechanisms. Thereafter, the present study investigated the effects of atorvastatin and rosuvastatin on proliferation, DNA synthsis and apoptosis of cultured neonatal rat cardiac fibroblasts with chromatometry, flow cytometry and invert microscope, fluorescence microscope. In the last section, the proliferation promotion and apoptosis induction effects of atorvastatin was compared with rosuvastatin. The study provided new ideas and experimental evidences for anti-myocardial fibrosis therapy. The present study included five parts as following:Part 1 The effects of aldosterone on proliferation and collagen synthesis in cultured neonatal cardiac fibroblastsObjective: To investigate the effects of Ald on proliferation and collagen synthesis in cultured neonatal cardiac fibroblasts.Method: In cultured cardiac fibroblasts of neonatal Sprague-Dawley (S-D) rats, the effects of Ald and Spi on proliferation were measured by MTT colorimetric assay, the effects of Ald and Spi on DNA synthesis were measured by Brdu colorimetric assay, the effects of Ald and Spi on cell cycle were measured by flow cytometry technique, synthesis of collagen was observed by the hydroxyproline concentration determined, p-ERK1/2, p-AKT, Cyclin D1, Cyclin E2 level were tested by immunofluorescence technique.Results: 1. Effects of Ald and Spi on proliferation of CFs: Compared with the control group, 10-9 mol/L 10-7 mol/L Ald significantly increased the proliferation of fibroblasts respectively (P <0.01, P <0.05, P < 0.05), 10-10 mol/L Ald didn't increased the proliferation of fibroblasts, and there were no difference among those four groups. Compared with 10-9 mol/L, 10-8 mol/L, 10-7 mol/L Ald groups, the proliferation of fibroblasts in 10-9 mol/L, 10-8 mol/L, 10-7 mol/L Ald +10-6 mol/L Spi groups were significantly decreased respectively (P <0.01, P<0.05, P<0.05) , which were not different from the control group.2. Effects of Ald and Spi on DNA synthesis of CFs: Compared with the control group, 10-9 mol/L 10-7 mol/L Ald significantly increased the DNA synthesis of fibroblasts respectively (All P <0.01). DNA synthesis in 10-7 mol/L Ald group was more than 10-8 mol/L Ald group (P <0.01), but there were no difference between 10-8 mol/L and 10-9 mol/L Ald groups. Compared with 10-9 mol/L, 10-8 mol/L, 10-7 mol/L Ald groups, DNA synthesis of fibroblasts in 10-9 mol/L, 10-8 mol/L, 10-7 mol/L Ald +10-6 mol/L Spi groups were significantly decreased respectively (All P <0.01) , which were not different from the control group.3. Effects of Ald and Spi on cell cycle change of CFs: 1) G1 phase cell proportion: Compared with the control group, 10-7mol/L Ald significantly decreased the proportion of fibroblasts in G1 phase at 24 h, 48 h, 72 h respectively (All P <0.01). The proportion of fibroblasts in G1 phase in 10(-7mol/L Ald 48 h group was less than 10-7 mol/L Ald 24 h group, and 10(-7mol/L Ald 72 h group was less than 10-7 mol/L Ald 48 h group(P <0.01, P <0.01). Compared with 10-7mol/L Ald groups, the proportion of fibroblasts in G1 phase in 10-7 mol/L Ald +10-6 mol/L Spi groups were significantly increased at 24 h, 48 h, 72 h respectively (All P <0.01).2 S phase cell proportion: Compared with the control group, 10(-7mol/L Ald significantly increased the proportion of fibroblasts in S phase at 24 h, 48 h, 72 h respectively (All P <0.01). The proportion of fibroblasts in S phase in 10-7 mol/L Ald 48 h group was more than 10(-7 mol/L Ald 24 h group, and 10-7 mol/L Ald 72 h group was more than 10(-7 mol/L Ald 48 h group(P <0.01, P <0.01). Compared with 10(-7mol/L Ald groups, the proportion of fibroblasts in S phase in 10(-7mol/L Ald +10-6 mol/L Spi 24 h, 48 h, 72 h groups were significantly decreased respectively (All P <0.01).3 PI: Compared with the control group, 10-7 mol/L Ald significantly increased PI of fibroblasts at 24 h, 48 h, 72 h respectively (All P <0.01). PI of fibroblasts in 10-7 mol/L Ald 48 h group was more than 10-7 mol/L Ald 24 h group, and 10-7 mol/L Ald 72 h group was more than 10-7 mol/L Ald 48 h group(P <0.01, P <0.01). Compared with 10-7 mol/L Ald groups, PI of fibroblasts in 10-7 mol/L Ald +10-6 mol/L Spi 24 h, 48 h, 72 h groups were significantly decreased respectively (All P <0.01).4. Effects of Ald and Spi on collagen synthesis in CFs: Compared with the control group, 10-7 mol/L Ald significantly increased the collagen synthesis in fibroblasts at 24 h, 48 h, 72 h respectively (All P <0.01). Collagen synthesis in 10-7 mol/L Ald 48 h group was more than 10(-7 mol/L Ald 24 h group, 10-7 mol/L Ald 72 h group was more than 10(-7 mol/L Ald 48 h group (P <0.01, P <0.01). Compared with 10-7 mol/L Ald 24 h, 48 h, 72 h groups, collagen synthesis in fibroblasts in correspondent 10-7 mol/L Ald +10-6 mol/L Spi groups were significantly decreased respectively (All P <0.01) .5. Effects of Ald on expression of Cyclin D1 in CFs: Compared with the control group, 10-7 mol/L Ald significantly increased the expression of Cyclin D1 in fibroblasts nucleus at 24 h, 48 h respectively (P <0.01, P <0.05). The expression of Cyclin D1 in 10-7 mol/L Ald 24 h group was higher than 10-7 mol/L Ald 48 h group.6. Effects of Ald on expression of Cyclin E2 in CFs: Compared with the control group, 10-7 mol/L Ald significantly increased the expression of Cyclin E2 in fibroblasts nucleus at 24 h, 48 h, 72 h respectively (P <0.01, P <0.05, P <0.05). The expression of Cyclin E2 in 10-7 mol/L Ald 24 h group was higher than 10-7 mol/L Ald 48 h and 72 h groups.7. Effects of Ald on expression of P-ERK1/2 in CFs: Compared with the control group, 10-7 mol/L Ald significantly increased the expression of P-ERK1/2 in fibroblasts at 5 min, 15 min, 30 min, 60 min, 2 h, 4 h, 8 h, 24 h respectively (All P <0.05). The expression of P-ERK1/2 in 10-7 mol/L Ald 30 min and 4 h group were the highest among all groups.8. Effects of Ald on expression of p-AKT in CFs: Compared with the control group, 10-7 mol/L Ald significantly increased the expression of p-AKT in fibroblasts at 45 min, 60 min, 2 h respectively (P <0.01, P <0.01, P <0.01). But there was not significant peak amplitude.Conclusion: Exogenous aldosterone stimulated cardiac fibroblasts proliferation, DNA synthesis and collagen synthesis via activating P-ERK1/2 and p-AKT. Ald increased CFs proliferation by upregulating Cycin D1 and Cyclin E2 expression. Part 2 The effects of atorvastatin on aldosterone induced proliferation and collagen synthesis in cultured neonatal cardiac fibroblastsObjective: To investigate the effects of Ato on Ald induced proliferation and collagen synthesis in cultured neonatal cardiac fibroblasts.Method: In cultured cardiac fibroblasts of neonatal Sprague-Dawley (S-D) rats, the effects of Ato on Ald induced proliferation were measured by MTT colorimetric assay, the effects of Ato on Ald induced DNA synthesis were measured by Brdu colorimetric assay, the effects of Ato on Ald induced cell cycle change were measured by flow cytometry technique, synthesis of collagen was observed by the hydroxyproline concentration determined, p-ERK1/2, p-AKT, Cyclin D1, Cyclin E2 level were tested by immunofluorescence technique and Western Blot.Results: 1. Effects of Ato on Ald induced proliferation of CFs: Compared with the control group, 10-7 mol/L Ald significantly increased the proliferation of fibroblasts (P <0.01). Compared with 10-7 mol/L Ald group, the proliferation of fibroblasts in 10-7 mol/L Ald +10-6 mol/L Spi, 10-8 mol/L 10-5 mol/L Ato groups were significantly decreased respectively (All P <0.01). There were not difference between control group and 10-7 mol/L Ald +10-6 mol/L Spi, 10-8 mol/L, 10-7 mol/L Ato groups, however, the proliferation of fibroblasts in 10-7 mol/L Ald +10(-6 mol/L, 10-5 mol/L, Ato groups were lower than control group (P <0.01, P <0.01). Trere were not difference between 10-7 mol/L Ald+10-8 mol/L Ato group and 10-7 mol/L Ald+10-7 mol/L Ato group, 10-7 mol/L Ald+10(-6 mol/L Ato group and 10-7 mol/L Ald+10-5 mol/L Ato group. The proliferation of fibroblasts in 10-7 mol/L Ald+10-6 mol/L Ato group was lower than 10-7 mol/L Ald+10-7 mol/L Ato group. Trere were no difference between 10-7 mol/L Ald+10-6 mol/L Spi group and 10-7 mol/L Ald+10-8 mol/L, 10-7 mol/L Ato groups. The proliferation of fibroblasts in 10-7 mol/L Ald+10-6 mol/L, 10-5 mol/L Ato groups was lower than 10(-7 mol/L Ald+10-6 mol/L Spi group.2. Effects of Ato on Ald induced DNA synthsis of CFs: Compared with the control group, 10-7 mol/L Ald significantly increased DNA synthsis of fibroblasts (P <0.01). Compared with 10-7 mol/L Ald group, DNA synthesis of fibroblasts in 10-7mol/L Ald +10(-6mol/L Spi, 10-8 mol/L 10-5 mol/L Ato groups were significantly decreased respectively (All P <0.01). Trere were not difference between control group and 10-7 mol/L Ald+10(-6mol/L Spi, 10-8 mol/L, 10-7 mol/L Ato groups, however, DNA synthesis of fibroblasts in 10(-7mol/L Ald +10-6 mol/L, 10-5 mol/L Ato groups were lower than control group (P <0.01, P <0.01). DNA synthesis of fibroblasts in 10-7 mol/L Ald+10-7 mol/L Ato group was lower than 10(-7 mol/L Ald+10-8 mol/L Ato group, DNA synthesis of fibroblasts in 10(-7 mol/L Ald+10-6 mol/L Ato group was lower than 10-7 mol/L Ald+10(-7 mol/L Ato group, DNA synthesis of fibroblasts in 10-7 mol/L Ald+10(-5 mol/L Ato group was lower than 10-7mol/L Ald+10-6 mol/L Ato group(All P <0.01).3. Effects of Ato on Ald induced cell cycle change of CFs: 1 S phase cell proportion: Compared with the control group, 10-7 mol/L Ald significantly increased the proportion of fibroblasts in S phase at 24 h, 48 h, 72 h respectively (All P <0.01). Compared with 10-7 mol/L Ald group, the proportion of fibroblasts in S phase in 10-7 mol/L Ald+10-6 mol/L Spi, 10-7 10-5 mol/L Ato 24 h, 48 h, 72 h groups were significantly decreased respectively (All P <0.01).2 PI: Compared with the control group, 10-7 mol/L Ald significantly increased PI of fibroblasts at 24 h, 48 h, 72 h respectively (All P <0.01). Compared with 10-7 mol/L Ald groups, PI of fibroblasts in 10-7 mol/L Ald +10-6 mol/L Spi, 10(-7 10-5 mol/L Ato 24 h, 48 h, 72 h groups were significantly decreased respectively (All P <0.01).4. Effects of Ato on Ald induced collagen synthesis increase in CFs: Compared with the control group, 10-7 mol/L Ald significantly increased the collagen synthesis in fibroblasts at 24 h, 48 h, 72 h respectively (All P <0.01). Compared with 10-7 mol/L Ald 24 h, 48 h, 72 h groups, collagen synthesis in fibroblasts in 10-7 mol/L Ald+10-6 mol/L Spi, 10(-7 10-5 mol/L Ato 24 h, 48 h, 72 h groups were significantly decreased respectively (All P <0.01).5. Effects of Ato on Ald induced expression of Cyclin D1 in CFs: Compared with 10-7mol/L Ald 24 h group, 10-7 mol/L Ald +10-5 mol/L Ato significantly inhibited the expression of Cyclin D1 in fibroblasts nucleus at 24 h (P <0.01).6. Effects of Ato on Ald induced expression of Cyclin E2 in CFs: Compared with 10-7 mol/L Ald 24 h group, 10-7 mol/L Ald +10-5 mol/L Ato significantly inhibited the expression of Cyclin E2 in fibroblasts nucleus at 24 h (P <0.01).7. Effects of Ato on Ald induced expression of p-ERK1/2 in CFs: 1 There were not difference between 10-7 mol/L Ald 30 min group and 10-7 mol/L Ald +10-7 10-5 mol/L Ato 30 min groups on expression of p-ERK1/2 in CFs.2 Compared with 10-7 mol/L Ald 4 h group, 10-7 mol/L Ald +10-7 10-5 mol/L Ato significantly inhibited the expression of p-ERK1/2 in fibroblasts at 4 h (All P <0.01). P-ERK1/2 expression in fibroblasts in 10-7 mol/L Ald +10-6 mol/L Ato 4 h group was lower than 10-7 mol/L Ald+10-7 mol/L Ato 4 h group (P <0.01).3 Western Blot results show: compared with 10-7 mol/L Ald 4 h group, 10-7 mol/L Ald +10-7 10-5 mol/L Ato significantly inhibited the expression of p-ERK1/2 in fibroblasts at 4 h (P <0.01, P <0.01, P <0.01).8. Effects of Ato on Ald induced expression of p-AKT in CFs: Compared with 10-7 mol/L Ald 1 h group, 10-7 mol/L Ald +10-7 10-5 mol/L Ato significantly inhibited the expression of p-AKT in fibroblasts at 1 h (All P <0.01). p-AKT expression in fibroblasts in 10-7 mol/L Ald+10-6 mol/L Ato 1 h groups was lower than 10-7 mol/L Ald+10-7 mol/L Ato 1 h groups, p-AKT expression in fibroblasts in 10-7 mol/L Ald+10-5 the collagen synthesis in fibroblasts at 24 h, 48 h, 72 h respectively (All P <0.01). Compared with 10-7 mol/L Ald 24 h, 48 h, 72 h groups, collagen synthesis in fibroblasts in 10-7 mol/L Ald+10-6 mol/L Spi, 10-7～10-5 mol/L Ato 24 h, 48 h, 72 h groups were significantly decreased respectively (All P <0.01).5. Effects of Ato on Ald induced expression of Cyclin D1 in CFs: Compared with 10-7mol/L Ald 24 h group, 10-7 mol/L Ald +10-5 mol/L Ato significantly inhibited the expression of Cyclin D1 in fibroblasts nucleus at 24 h (P <0.01).6. Effects of Ato on Ald induced expression of Cyclin E2 in CFs: Compared with 10-7 mol/L Ald 24 h group, 10-7 mol/L Ald +10-5 mol/L Ato significantly inhibited the expression of Cyclin E2 in fibroblasts nucleus at 24 h (P <0.01).7. Effects of Ato on Ald induced expression of p-ERK1/2 in CFs: 1 There were not difference between 10-7 mol/L Ald 30 min group and 10-7 mol/L Ald +10-7～10-5 mol/L Ato 30 min groups on expression of p-ERK1/2 in CFs.2 Compared with 10-7 mol/L Ald 4 h group, 10-7 mol/L Ald +10-7 10-5 mol/L Ato significantly inhibited the expression of p-ERK1/2 in fibroblasts at 4 h (All P <0.01). P-ERK1/2 expression in fibroblasts in 10-7 mol/L Ald +10-6 mol/L Ato 4 h group was lower than 10-7 mol/L Ald+10-7 mol/L Ato 4 h group (P <0.01).3 Western Blot results show: compared with 10-7 mol/L Ald 4 h group, 10-7 mol/L Ald +10-7～10-5 mol/L Ato significantly inhibited the expression of p-ERK1/2 in fibroblasts at 4 h (P <0.01, P <0.01, P <0.01).8. Effects of Ato on Ald induced expression of p-AKT in CFs: Compared with 10-7 mol/L Ald 1 h group, 10-7 mol/L Ald +10-7～10-5 mol/L Ato significantly inhibited the expression of p-AKT in fibroblasts at 1 h (All P <0.01). p-AKT expression in fibroblasts in 10-7 mol/L Ald+10-6 mol/L Ato 1 h groups was lower than 10-7 mol/L Ald+10-7 mol/L Ato 1 h groups, p-AKT expression in fibroblasts in 10-7 mol/L Ald+10-5 10-5 mol/L Ato +10-5 mol/L FPP groups were significantly increased respectively (All P <0.01). 10-5 mol/L FPP could not reverse the inhibition effects of 10-5 mol/L Ato on CFs DNA synthesis completely. The inhibition effects of 10-7 mol/L Ato group was higher than 10-8mol/L Ato group (P <0.01).3. Effects of Ato on morphology of CFs: 1 Observed with invert microscope, the shapes of fibroblasts without treated with Ato were regular, fusiform and polygon, intercellular space was small and anchoring growing. Fibroblasts in Ato group were shrunk and small. Intercellular space became large, and the ability of anchoring was reduced. The morphology of fibroblasts was changed more prominent, with the dose and time increase after Ato administration.2 Observed with invert microscope after Wright ' s- Giemsa staining, the shapes of fibroblasts without treated with Ato were regular, fusiform and polygon, the endochylema were stained with uniform light pink, the nucleus were big and stained with deep pink or light purple, intercellular space was small and anchoring growing. Fibroblasts in Ato group were shrunk and small, with many pseudopodia. Intercellular space became large, nucleus became pyknosis and dark stained, and the ability of anchoring was reduced. The morphology of fibroblasts was changed more prominent, with the dose and time increase after Ato administration.3 Observed with fluorescence microscope after Hochest33258 staining, the nucleus without treated with Ato were regular, oval-shap and stained with light blue fluorescence. The nucleus in Ato group became pyknosis, disfiguration or disintegration, and dark stained. The morphology of nucleus was changed more prominent, with the dose and time increase after Ato administration.4. Effects of Ato on apoptosis of CFs detecting by flow cytometry technique with AnexinV/PI double staining: 1 Analysis of early apoptosis fibroblasts proportion: Compared with the correspondent control groups, 10-7 mol/L 10-5 mol/L Ato significantly increased the proportion of early apoptosis fibroblasts at 24 h, 48 h, 72 h respectively (All P <0.01). Ato could increase early apoptosis fibroblasts proportion by time and dosage, except between 10-7 mol/L Ato 24h and 10-6 mol/L Ato 24h, 10-7 mol/L Ato 72h and 10-6 mol/L Ato 72h.2 Analysis of late apoptosis fibroblasts proportion: Compared with the correspondent control groups, 10-7 mol/L 10-5 mol/L Ato significantly increased the proportion of late apoptosis fibroblasts at 24 h, 48 h, 72 h respectively (All P <0.01). Ato could increase late apoptosis fibroblasts proportion by time and dosage, except between 10-7 mol/L Ato 24 h and 10-7 mol/L Ato 48 h group, 10-6 mol/L Ato 24 h and 10-6 mol/L Ato 48 h group.3 Analysis of total apoptosis fibroblasts proportion: Compared with the correspondent control group, 10-7 mol/L 10-5 mol/L Ato significantly increased the proportion of total apoptosis fibroblasts at 24 h, 48 h, 72 h respectively (All P <0.01). Ato could increase total apoptosis fibroblasts proportion by time and dosage.5. Effects of Ato on apoptosis of CFs detecting sub-G1 by flow cytometry technique with PI staining: Compared with the correspondent control groups, 10-7 mol/L 10-5 mol/L Ato significantly increased the proportion of apoptosis fibroblasts at 24 h, 48 h, 72 h (All P <0.01). Compared with 10-5 mol/L Ato groups, the proportion of apoptosis fibroblasts in correspondent 10-5 mol/L Ato +10-5mol/L FPP 24 h, 48 h, 72 h groups were significantly increased respectively (All P <0.01). Ato could increase apoptosis fibroblasts proportion by time and dosage (All P <0.05).Conclusion: Ato could inhibit proliferation and DNA synthesis of CFs and induce CFs apoptosis. Part 4 The effects of rosuvastatin on proliferation inhibition and apoptosis induction of cultured neonatal cardiac fibroblastsObjective: To investigate the effects of Ros on proliferation inhibition and apoptosis induction of cultured neonatal cardiac fibroblasts.Method: In cultured cardiac fibroblasts of neonatal Sprague-Dawley (S-D) rats, the effects of Ros on proliferation were measured by MTT colorimetric assay, the effects of Ros on DNA synthesis were measured by Brdu colorimetric assay, the effects of Ros on apoptosis were observed by inverted biological microscope and fluorescence microscope, and measured by flow cytometry technique.Results: 1. Effects of Ros on proliferation of CFs: Compared with the control group, 10-7 mol/L 10-5 mol/L Ros significantly inhibited the proliferation of fibroblasts respectively (All P <0.01). Compared with 10-7 mol/L 10-5 mol/L Ros groups, the proliferation of fibroblasts in 10-7 mol/L 10-5 mol/L Ros +10-5 mol/L FPP groups were significantly increased respectively (All P <0.01). 10-5 mol/L FPP could not reverse the inhibition effects of CFs proliferation in 10-5 mol/L Ros group completely. The inhibition effects of 10-7 mol/L Ros was higher than 10-8 mol/L Ros, and the inhibition effects of 10-6 mol/L Ros was higher than 10-7 mol/L Ros (P <0.01, P <0.01).2. Effects of Ros on DNA synthesis of CFs: Compared with the control group, 10-7 mol/L 10-5 mol/L Ros significantly inhibited DNA synthesis of fibroblasts respectively (All P <0.01). Compared with 10-7 mol/L 10-5 mol/L Ros groups, DNA synthesis of fibroblasts in 10-7 mol/L 10-5 mol/L Ros +10-5 mol/L FPP groups were significantly increased respectively (All P <0.01). 10-5 mol/L FPP could not reverse the inhibition effects on CFs DNA synthesis in 10-5 mol/L Ros group completely. The inhibition effects of 10-7 mol/L Ros was higher than 10-8 mol/L Ros, and the inhibition effects of 10-5 mol/L Ros group was higher than 10-6 mol/L Ros (P <0.01, P <0.01).3. Effects of Ros on morphology of CFs: 1 Observed with invert microscope, the shapes of fibroblasts without treated with Ros were regular, fusiform and polygon, intercellular space was small and anchoring growing. Fibroblasts in Ros group were shrunk and small. Intercellular space became large, and the ability of anchoring was reduced. The morphology of fibroblasts was changed more prominent, with the dose and time increase after Ros administration.2 Observed with invert microscope after Wright's- Giemsa staining, the shapes of fibroblasts without treated with Ros were regular, fusiform and polygon, the endochylema were stained with uniform light pink, the nucleus were big and stained with deep pink or light purple, intercellular space was small and anchoring growing. Fibroblasts in Ros group were shrunk and small, with many pseudopodia. Intercellular space became large, nucleus became pyknosis and dark stained, and the ability of anchoring was reduced. The morphology of fibroblasts was changed more prominent, with the dose and time increase after Ros administration.3 Observed with fluorescence microscope after Hochest33258 staining, the nucleus without treated with Ros were regular, oval-shap and stained with light blue fluorescence. The nucleus in Ros group became pyknosis, disfiguration or disintegration, and dark stained. The morphology of nucleus was changed more prominent, with the dose and time increase after Ros administration.4. Effects of Ros on apoptosis of CFs detecting by flow cytometry technique with AnexinV/PI double staining: 1 Analysis of early apoptosis fibroblasts proportion: Compared with the correspondent control group, 10-7 mol/L 10-5 mol/L Ros significantly increased the proportion of early apoptosis fibroblasts at 24 h, 48 h, 72 h respectively (All P <0.05), except 10-7 mol/L Ros 24 h group. Ros could increase early apoptosis fibroblasts proportion by time and dosage (All P <0.05).2 Analysis of late apoptosis fibroblasts proportion: Compared with the correspondent control group, 10-7 mol/L 10-5 mol/L Ros significantly increased the proportion of late apoptosis fibroblasts at 24 h, 48 h, 72 h respectively (All P <0.05) , except 10-7 mol/L Ros 24 h group. Ros could increase late apoptosis fibroblasts proportion by time and dosage (All P <0.05).3 Analysis of total apoptosis fibroblasts proportion: Compared with the correspondent control group, 10-7 mol/L 10-5 mol/L Ros significantly increased the proportion of total apoptosis fibroblasts at 24 h, 48 h, 72 h respectively (All P <0.05). Ros could increase total apoptosis fibroblasts proportion by time and dosage.5. Effects of Ros on apoptosis of CFs detecting sub-G1 by flow cytometry technique with PI staining: Compared with the correspondent control group, 10-7 mol/L 10-5 mol/L Ros significantly increased the proportion of apoptosis fibroblasts at 24 h, 48 h, 72 h respectively (All P <0.01). Compared with 10-5 mol/L Ros groups, the proportion of apoptosis fibroblasts in correspondent 10-5 mol/L Ros +10-5 mol/L FPP 24 h, 48 h, 72 h groups were significantly increased respectively (All P <0.01). Ros could increase apoptosis fibroblasts proportion by time and dosage (All P <0.05).Conclusion: Ros could inhibit proliferation and DNA synthesis of CFs and induce CFs apoptosis.Part 5 Comparision of Effects on cardiac fibroblasts proliferation and apoptosis between Rosuvastatin and AtorvatatinObjective: To compare the effects of Ato and Ros on proliferation and apoptosis of cultured neonatal cardiac fibroblasts.Method: In cultured cardiac fibroblasts of neonatal Sprague-Dawley (S-D) rats, the effects of Ato and Ros on proliferation were measured by MTT colorimetric assay, the effects of Ato and Ros on DNA synthesis were measured by Brdu colorimetric assay, the effects of Ato and Ros on cell cycle were measured by flow cytometry technique, synthesis of collagen was observed by the hydroxyproline concentration determined, and apoptosis were measured by flow cytometry technique.Results: 1. Effects of Ato and Ros on proliferation of CFs: Compared with the control group, 10-7 mol/L 10-5 mol/L Ato and Ros significantly inhibited the proliferation of fibroblasts respectively (All P <0.01). The inhibition effects of Ato and Ros groups was equal to each other.2. Effects of Ato and Ros on DNA synthesis of CFs: Compared with the control group, 10-7 mol/L 10-5 mol/L Ato and Ros significantly inhibited the DNA synthesis in fibroblasts respectively (All P <0.01). The inhibition effects of Ato and Ros groups was equal to each other.3. Effects of Ato and Ros on cell cycle of CFs: Compared with the control group, 10-7 mol/L 10-5 mol/L Ato and Ros significantly increased cell proportion in G0/G1 phase, and decreased cell proportion in S phase and PI (All P <0.01). There were not difference between Ato and Ros groups on cell cycle inhibition.4. Effects of Ato and Ros on apoptosis of CFs detecting sub-G1 by flow cytometry technique with PI staining: Compared with the correspondent control group, 10-7 mol/L 10-5 mol/L Ato and Ros significantly increased the proportion of apoptosis fibroblasts at 24 h, 48 h, 72 h respectively (All P <0.01). There were not difference between Ato and Ros groups on apoptosis induction.5. Effects of Ato and Ros on collagen synthesis in CFs: Compared with the control group, 10-7 mol/L 10-5 mol/L Ato and Ros significantly inhibited the collagen synthesis in fibroblasts respectively (All P <0.01). The inhibition effects of Ato and Ros groups was equal to each other.Conclusion: Both Ato and Ro are effective antifibrotic agents. And their effects are equal to each other. |
PDF Full Text Request