Inhibition of neointimal hyperplasia in rats treated with atorvastatin after carotid artery injury may be mainly associated with down-regulation of survivin and Fas expression

Figures & data

Figure 1. The intimal hyperplasia after vascular injury. A1, 7 d after vascular injury in group without treatment with significant intimal hyperplasia, right carotid artery. B1, 7 d after vascular injury and with 10 mg/kg atorvastatin treatment group with less intimal hyperplasia, right carotid artery. A2, 14 d after vascular injury in group without atorvastatin treatment with significant intimal hyperplasia, right carotid artery. B2, 14 d after vascular injury and with 10 mg/kg atorvastatin treatment group, right carotid artery, less intimal hyperplasia than without treatment group. A3, 28 d after vascular injury in group without atorvastatin treatment with significant intimal hyperplasia, right carotid artery. B3, 28 d after vascular injury in group with 10 mg/kg atorvastatin treatment, right carotid artery, less intimal hyperplasia than group without treatment. The directions of arrow are neointima.

Table 1. The change of intimal media and intimal/media I/M ratio after vascular injury with or without treatment with atorvastatin.

Groups	Intimal (μm)	Media (μm)	Intimal/media (I/M)
Normal control group	1.01 ± 0.146^a	31.39 ± 2.485	0.0324 ± 0.0054^a
Sham operation group	1.00 ± 0.105^b	31.10 ± 1.740	0.0322 ± 0.0034^b
Without treatment group: 3 d	8.42 ± 0.449	32.8 ± 2.115	0.2571 ± 0.0155
Without treatment group: 7 d	31.87 ± 3.10^c	40.83 ± 1.369	0.7804 ± 0.0697^c
Without treatment group: 14 d	41.58 ± 1.64^d	39.87 ± 1.297	1.0438 ± 0.0522^d
Without treatment group: 28 d	28.38 ± 2.59^e	40.19 ± 2.250	0.7061 ± 0.0569^e
Treatment group: 3 d	8.195 ± 0.40	34.72 ± 1.245	0.2363 ± 0.0158
Treatment group: 7 d	13.72 ± 1.25	40.62 ± 1.147	0.3382 ± 0.0371
Treatment group: 14 d	12.30 ± 0.99	39.91 ± 0.469	0.3082 ± 0.0241
Treatment group: 28 d	13.62 ± 1.56	39.98 ± 1.612	0.3401 ± 0.0269

^aCompared with vascular injury groups with or without treatment, p < 0.01.

^bCompared with vascular injury groups with or without treatment, p < 0.01.

^cCompared with the 7 d treatment group, p < 0.01.

^dCompared with the 14 d treatment group, p < 0.01.

^eCompared with the 28 d treatment group, p < 0.01.

Figure 2. The results of TUNEL staining. A1, 2, 3, and 4 are 3, 7, 14, and 28  d after vascular injury in groups without treatment, respectively. We can see significant intimal hyperplasia, and less TUNEL-positive cells. B1, 2, 3, and 4 are 3, 7, 14, and 28 d after vascular injury in groups with atorvastatin treatment, respectively. We can see less intimal hyperplasia and more TUNEL-positive cells, indicated that atorvastatin can inhibit neointimal formation and induce SMCs apoptosis.

Table 2. The changes of levels of survivin and sFas in the plasma of rats after treated with atorvastatin.

Groups	Survivin (pg/ml)	sFas (pg/ml)
Normal control group	230.00 ± 62.78^a	1142.80 ± 155.66^a
Sham operation group	276.00 ± 49.48^b	1536.40 ± 365.37^b
3 d without treatment group	464.80 ± 105.27	3256.00 ± 478.20
7 d without treatment group	998.00 ± 146.70	4362.00 ± 639.92
14 d without treatment group	1089.20 ± 232.32	2787.00 ± 403.63
28 d without treatment group	562.00 ± 90.11	2148.00 ± 257.14
3 d atorvastatin treatment group	541.60 ± 132.37	2278.00 ± 428.28
7 d atorvastatin treatment group	501.60 ± 107.25^c	2054.00 ± 264.16^c
14 d atorvastatin treatment group	286.00 ± 52.27^d	1838.00 ± 214.52^d
28 d atorvastatin treatment group	218.80 ± 33.24^e	1580.00 ± 233.13^e

^a,bCompared with 3, 7, 14, and 28 d after surgery but without treatment groups and 3and 7 treatment groups, p < 0.01.

^c,d,eCompared with 7, 14, and 28 d after surgery but without treatment groups, p < 0.01.

Figure 3. The expression of Fas in local vascular injury sites. (A) Relative levels of mRNAs encoding Fas in local vascular injury sites. Lane 1, β-actin. Lane 2, normal control artery with no Fas mRNA expression. Lanes 3, 4, and 5 are 28, 14, and 7 d after vascular injury and with atorvastatin treatment groups, respectively, there are Fas mRNA expressions. Lanes 6, 7, 8, and 9 is 28, 14, 7, and 3 d after vascular injury but without atorvastatin treatment, respectively, we can also see various types of Fas mRNA expression. Lane 10, marker. (B) Relative level of Fas expression was analyzed by Western Blotting. (B1) Tubulin, as an internal standard; (B2) Fas western blotting analysis. Lanes 1, 2, and 3 are 28, 14, and 7 d after vascular injury and with atorvastatin treatment groups, respectively. Lanes 4, 5, 6, and 7 are 28, 14, 7, and 3  d after vascular injury but without atorvastatin treatment, respectively. Fas protein expression is similar to the Fas mRNA expression.

Figure 4. The expression of survivin in local vascular injury site. (A) Relative levels of mRNAs encoding survivin expression. Lane 1, blank control. Lane 2, the normal control group, no survivin mRNA expression. Lanes 3 and 4 were β-actin. Lanes 5 and 6 is 28 and 14 d after vascular injury in group without treatment, respectively, these are survivin mRNA expression. Lane 7 is 7 d after vascular injury in group without treatment, no survivin mRNA expression. Lanes 8 and 9 is 28 and 14 d after vascular injury in groups with atorvastatin treatment, respectively, no survivin mRNA expression. Lane 10, marker. (B) Relative level of survivin expression was analyzed by Western blotting. (B1) Tubulin, as an internal standard; (B2) survivin protein. Lanes 1, 2, and 3 are 28, 14, and 7  d after vascular injury in group without treatment, respectively. Lanes 4 and 5 are 28 and 14 d after vascular injury in group with atorvastatin treatment. Survivin expression is similar to the survivin mRNA.