Effect of free creatine therapy on cisplatin-induced renal damage

Figures & data

Table 1. Sequence of PCR primers.

Primers	Sequence of primers
36B4 Forward	5′-TCCGTATTGCAGAGTCCTTG-3′
36B4 Reverse	5′-GCACATCGCTCTGAGGAA-3′
D-loop Forward	5′-GGTTCTTACTTCAGGGCCATCA-3′
D-loop Reverse	5′-GATTAGACCCGTTACCATCGAGAT-3′
mtDNA^4834 Deletion Forward	5′-AAGGACGAACCTGAGCCCTAATA-3′
mtDNA^4834 Deletion Reverse	5′-CGAAGTAGATGATCCGTATGCTGTA-3′

Table 2. QPCR protocols of 36B4, D-loop, mtDNA⁴⁸³⁴ deletion, and telomere.

Step	36B4	D-loop	mtDNA^4834 deletion
Activation	10 min at 95 °C	10 min at 95 °C	10 min at 95 °C
Denaturation	5 s at 95 °C	5 s at 95 °C	5 s at 95 °C
Annealing	10 s at 56 °C	10 s at 56 °C	10 s at 56 °C
Elongation	6 s at 72 °C	6 s at 72 °C	3 s at 72 °C
Cycle number	40	45	45

Table 3. Weight alterations of rats through experiment.

Day	Group I	Group II	Group III	p Values	Groups' p values
7th day	−44.7 ± 19.43 * −37 (9.8%) (−87, −28)	1.3 ± 17.93 * −1.5 (0.4%) (−22, 29)	2 ± 17.6 * 2 (0.5%) (−30, 24)	<0.001	I–II: <0.001 I–III: <0.001 II–III: 0.931
30th day	−50.6 ± 34.54 * −59 (16%) (−99, 10)	−13.5 ± 33.29 * −11.5 (3%) (–86, 30)	12.7 ± 27.08 * 18 (5.6%) (−48, 44)	0.002	I–II: 0.035 I–III: <0.001 II–III: 0.084

Note: Values are expressed as mean ± Standard deviation * median (minimum, maximum).

Table 4. Kidney function tests alterations of rats through experiment.

Day		Group I	Group II	Group III	p Value	Groups' p values
7th day	BUN (mg/dL)	146.5 ± 42.21 * 137 (92−219)	65.89 ± 50.12 * 45 (16–176)	15.1 ± 2.13 * 15.5 (11–19)	<0.001	I–II: 0.002 I–III: <0.001 II–III: <0.001
	Creatinine (mg/dL)	6.88 ± 3.89 * 6 (2.36–14.78)	1.73 ± 1.2 * 1.35 (0.38–4.23)	0.25 ± 0.03 * 0.25 (0.22–0.3)	<0.001	I–II: <0.001 I–III: <0.001 II–III: <0.001
30th day	BUN (mg/dL)	44.7 ± 21.15 * 38 (26–84)	19.22 ± 3.67 * 20 (12–24)	16 ± 2.54 * 15.5 (12–20)	<0.001	I–II: <0.001 I–III: <0.001 I–III: 0.035
	Creatinine (mg/dL)	0.83 ± 0.47 * 0.63 (0.42–1.97)	0.43 ± 0.11 * 0.4 (0.27–0.63)	0.3 ± 0.06 * 0.29 (0.24–0.4)	<0.001	I–II: 0.002 I–III: <0.001 II–III: 0.008

Note: Values are expressed as mean ± Standard deviation * median (minimum–maximum).

Table 5. Common deletions alterations of rats through experiment.

Day	Group I	Group II	Group III	p Value	Groups' p values
7th day	26.18 ± 2.27 25.8 * (23.8–31.9)	23.83 ± 1.62 24.1 * (21–26.2)	20.62 ± 2.93 21.4 * (13.5–23.3)	<0.001	I–II: 0.017 I–III: <0.001 II–III: 0.004
30th day	27.69 ± 6.58 26.85 * (20.3–44.4)	24.26 ± 1.13 24 * (22.8–26.6)	21.88 ± 2.27 22.15 * (18–25.2)	0.006	I–II: 0.095 I–III: 0.007 II–III: 0.013

Notes: Values are expressed as mean ± Standard deviation * median (minimum–maximum).

When groups were evaluated in terms of progress, no significant difference was found in all three groups (p = 0.959, 0.594, and 0.173, respectively).

Figure 1. Representative PAS-staining sections in each group. (a) Group I at 7th day, (b) Group II at 7th day, (c) Group III at 7th day, (d) Group I at 30th day, (e) Group II at 30th day, (f) Group III at 30th day (200 × magnification).

Table 6. Histopathological scores alterations of rats through experiment.

Day	Group I	Group II	Group III	p Values	Groups' p values
7th day	2.86 ± 0.47 * 2.89 (1.9–3.62)	2.31 ± 0.44 * 2.23 (1.8–3.2)	0.35 ± 0.17 * 0.31 (0.22–0.77)	<0.001	I–II: <0.028 I–III: <0.001 II–III: <0.001
30th day	3.05 ± 0.49 * 3 (2.4–3.64)	2.38 ± 0.79 * 2.57 (0.55–3.1)	0.59 ± 0.25 * 0.5 (0.33–1.12)	0.001	I–II: 0.046 I–III: <0.001 II–III: 0.001

Note: Values are expressed as mean ± Standard deviation * median (minimum–maximum).