Cardiol Res

Cardiology Research, ISSN 1923-2829 print, 1923-2837 online, Open Access

Article copyright, the authors; Journal compilation copyright, Cardiol Res and Elmer Press Inc

Journal website https://www.cardiologyres.org

Original Article

Volume 13, Number 6, December 2022, pages 339-356

Modeling Doxorubicin-Induced Cardiomyopathy With Fibrotic Myocardial Damage in Wistar Rats

Figure 3. Representative images of rat’s kidney of the control, DOX-25, DOX-20.4, and DOX-15 groups (stained by hematoxylin and eosin, × 25 (control group); × 50 (DOX-25, DOX-20.4, and DOX-15 groups)).

Figure 4. Dynamics of body weight in animals of the DOX-15, DOX-10, and DOX-5 groups. *Significant differences between the DOX-15 group of animals versus the DOX-10, DOX-5, and control groups (**P < 0.01, ***P < 0.001).

Figure 5. LVIDs, LVIDd and FS dynamics of animals of the DOX-15, DOX-10, and DOX-5 groups. *Dynamics compared to the initial value (e.g., in the DOX-15 group; *P < 0.05, **P < 0.01, ***P < 0.001). Δ: dynamics compared to the highest dose at each point of the experiment (e.g., in first month: DOX-5, control vs. DOX-15; Δ: P < 0.05, ΔΔ: P < 0.01, ΔΔΔ: P < 0.001). LVIDs: left ventricular end-systolic internal diameter; LVIDd: left ventricular end-diastolic internal diameter; FS: fractional shortening.

Figure 6. IVS and LVPW dynamics of animals of the DOX-15, DOX-10, and DOX-5 groups. *Dynamics compared to the initial value (*P < 0.05, **P < 0.01, ***P < 0.001). Δ: dynamics compared to the highest dose at each point of the experiment (e.g., in the second month: DOX-10, DOX-5 vs. DOX-15; Δ: P < 0.05, ΔΔ: P < 0.01). IVS: the dimensions of the anterior wall thickness; LVPW: the dimensions of the posterior wall thickness.

Figure 7. (a) Representative images of rat’s left cardiac ventricle of the control group; × 50; stained by hematoxylin and eosin. (b, c) Representative images of rat’s left cardiac ventricle of the DOX-15 group; stained by hematoxylin and eosin. (b) Dissociation of myocardial fibers is seen, forming cavities filled with blood (arrow); × 50. (c) Asterisks show vacuolization of the cytoplasm of cardiomyocytes, black arrows show nuclei of cardiocytes that undergo karyolysis, white arrows show hypertrophied fibers; × 25. (d) Representative images of rat’s left cardiac ventricle of the DOX-10 group; × 25; stained by hematoxylin and eosin. Anisonucleosis, medium and large cell vacuolization of the cytoplasm (arrows). (e) Representative images of rat’s left cardiac ventricle of the DOX-5 group; × 25; stained by hematoxylin and eosin. The arrows show vacuolization of the cytoplasm of cardiomyocytes, the absence of visible boundaries between individual cells.

Figure 9. The amount of collagen in the left cardiac ventricle with chronic doxorubicin-induced cardiomyopathy of rats of the control, DOX-15, DOX-10, and DOX-5 groups, %. Kruskal-Wallis test.

Figure 10. (a) Rat’s kidney of the Control group; × 25; stained by hematoxylin and eosin. (b, c) Rat’s kidney of the DOX-15 group; × 25; stained by hematoxylin and eosin. (b) The arrow shows the necrosis of the tubular epithelium, the sclerosed glomerulus shows in the center. (c) Massive tubular epithelial necrosis (arrow). (d) Rat’s kidney of the DOX-10 group; × 50; stained by hematoxylin and eosin. Global glomerulosclerosis of three renal glomeruli. (e) Rat’s kidney of the DOX-5 group; × 25; stained by hematoxylin and eosin. The black arrow shows necrosis of individual tubular epithelial cells. White arrows show thickening of the basement membrane of the glomerulus. Vacuolization of the cells of the parietal leaf of the capsule (asterisk).

Figure 11. Gene expression levels of fibrotic molecular markers in the model of chronic doxorubicin-induced cardiomyopathy. *P < 0.05, **P < 0.01, ***P < 0.001 compared to control. COL1A1, COL2A1, COL3A1: collagen, type I, II, III; ET-1: endothelin-1; FGF: fibroblast growth factors; MMP: matrix metalloproteinases; TGF-β: transforming growth factor-β; TIMP: tissue inhibitor of metalloprotease; TNF-α: tumor necrosis factor; α-SMA: alpha-smooth muscle actin.

**Table 1.** Brief Description of the Experimental Groups
Group, n	The route of administration	Dose	Frequency of administration	Cumulative dose
IP: intraperitoneally.
The first stage - modeling of acute doxorubicin cardiomyopathy
DOX-25 (n = 10)	IP	2.5 mg/kg	Ten times over 1.5 weeks given every day	25 mg/kg
DOX-20.4 (n = 10)	IP	3.4 mg/kg	Six times over 1.5 weeks given every other day	20.4 mg/kg
DOX-15 (n = 10)	IP	2.5 mg/kg	Six times over 1.5 weeks given every other day	15 mg/kg
Control (n = 5)	IP	1 mL 0.9% sodium chloride	Six times over 1.5 weeks given every other day	6 mL
The second stage - modeling of chronic doxorubicin cardiomyopathy
DOX-15 (n = 10)	IP	2.5 mg/kg	Six times over 2.5 weeks given in 2 days	15 mg/kg
DOX-10 (n = 10)	IP	1.67 mg/kg	Six times over 2.5 weeks given in 2 days	10 mg/kg
DOX-5 (n = 10)	IP	0.83 mg/kg	Six times over 2.5 weeks given in 2 days	5 mg/kg
Control (n = 10)	IP	1 mL 0.9% sodium chloride	Six times over 2.5 weeks given in 2 days	6 mL

Table 1. Brief Description of the Experimental Groups

Group, n

The route of administration

Dose

Frequency of administration

Cumulative dose

IP: intraperitoneally.

The first stage - modeling of acute doxorubicin cardiomyopathy

DOX-25 (n = 10)

2.5 mg/kg

Ten times over 1.5 weeks given every day

25 mg/kg

DOX-20.4 (n = 10)

3.4 mg/kg

Six times over 1.5 weeks given every other day

20.4 mg/kg

DOX-15 (n = 10)

2.5 mg/kg

Six times over 1.5 weeks given every other day

15 mg/kg

Control (n = 5)

1 mL 0.9% sodium chloride

Six times over 1.5 weeks given every other day

6 mL

The second stage - modeling of chronic doxorubicin cardiomyopathy

DOX-15 (n = 10)

2.5 mg/kg

Six times over 2.5 weeks given in 2 days

15 mg/kg

DOX-10 (n = 10)

1.67 mg/kg

Six times over 2.5 weeks given in 2 days

10 mg/kg

DOX-5 (n = 10)

0.83 mg/kg

Six times over 2.5 weeks given in 2 days

5 mg/kg

Control (n = 10)

1 mL 0.9% sodium chloride

Six times over 2.5 weeks given in 2 days

6 mL

**Table 2.** Lists of Primer Sequences Used for RT-qPCR
Genes	Primers	Sequences (5'-3')
α-SMA: alpha-smooth muscle actin; MMP: matrix metalloproteinases; TIMP: tissue inhibitor of metalloprotease; TGF: transforming growth factor; FGF: fibroblast growth factors; TNF: tumor necrosis factor; RT-qPCR: reverse transcription quantitative real time polymerase chain reaction.
ACTA (α-SMA)	Forward	CACCGCTGAACGTGAAATTG
	Reverse	CTTCTCCAGAGAGGAGGAAG
TGF-β1	Forward	GACTCTCCACCTGCAAGACC
	Reverse	GGACTGGCGAGCCTTAGTTT
FGF2	Forward	TCCATCAAGGGAGTGTGTGC
	Reverse	TCCGTGACCGGTAAGTGTTG
FGF4	Forward	CTACCTGCTGGGCCTCAAAA
	Reverse	CACACCCCGCTGCTGTC
Col1a1	Forward	GTGGATGGCTGCACGAGTC
	Reverse	GAGTTTGGGTTGTTGGTCTG
Col2a1	Forward	GCTGTGGAAGTGGATGAAGA
	Reverse	GAGGAACTGTGGAGAGACG
Col3a1	Forward	CAGGCCAATGGCAATGTAAAG
	Reverse	CATCCTCTAGAACTGTGTAAG
TNF-α	Forward	GGCTCCCTCTCATCAGTTC
	Reverse	CTGCTTGGTGGTTTGCTAC
ET-1 (endothelin-1)	Forward	TGATTCTCTTGCCTCTTCTTG
	Reverse	TATGGAATCTCCTGGCTCTC
TIMP-1	Forward	CTGAGAAGGGCTACCAGAG
	Reverse	GTCATCGAGACCCCAAGGT
TIMP-2	Forward	GGACCTGACAAGGACATCG
	Reverse	TTCTTTCCTCCAACGTCCAG
MMP-1	Forward	GATGAAAGGTGGACCAACAAT
	Reverse	CCAAGAGAATGGCCGAGTTC
MMP-2	Forward	TGGGGGAGATTCTCACTTTG
	Reverse	CCATCAGCGTTCCCATACTT
MMP-14	Forward	TGGGGTCATCTGCTTCTCTT
	Reverse	TAGGGCTCATATGCCCAAAG
Housekeeping genes	Primers	Sequences (5'- 3')
GAPDH	Forward	CAAGTTCAACGGCACAGTCA
	Reverse	CATACTCAGCACCAGCATCA
α-tubulin	Forward	CAATTCCATCCTCACCACC
	Reverse	CAACCTGTTTAAGTTAGTGTAG
TBP (TATA-box binding protein)	Forward	TGCGTTGATCTTCAGTTCTG
	Reverse	CTTGCTGCTAGTCTGGATTG
ACTB (β-actin)	Forward	GGTGTGATGGTGGGTATGG
	Reverse	GTTGGTGACAATGCCGTGTT

Table 2. Lists of Primer Sequences Used for RT-qPCR

Genes

Primers

Sequences (5'-3')

α-SMA: alpha-smooth muscle actin; MMP: matrix metalloproteinases; TIMP: tissue inhibitor of metalloprotease; TGF: transforming growth factor; FGF: fibroblast growth factors; TNF: tumor necrosis factor; RT-qPCR: reverse transcription quantitative real time polymerase chain reaction.

ACTA (α-SMA)

Forward

CACCGCTGAACGTGAAATTG

Reverse

CTTCTCCAGAGAGGAGGAAG

TGF-β1

Forward

GACTCTCCACCTGCAAGACC

Reverse

GGACTGGCGAGCCTTAGTTT

FGF2

Forward

TCCATCAAGGGAGTGTGTGC

Reverse

TCCGTGACCGGTAAGTGTTG

FGF4

Forward

CTACCTGCTGGGCCTCAAAA

Reverse

CACACCCCGCTGCTGTC

Col1a1

Forward

GTGGATGGCTGCACGAGTC

Reverse

GAGTTTGGGTTGTTGGTCTG

Col2a1

Forward

GCTGTGGAAGTGGATGAAGA

Reverse

GAGGAACTGTGGAGAGACG

Col3a1

Forward

CAGGCCAATGGCAATGTAAAG

Reverse

CATCCTCTAGAACTGTGTAAG

TNF-α

Forward

GGCTCCCTCTCATCAGTTC

Reverse

CTGCTTGGTGGTTTGCTAC

ET-1 (endothelin-1)

Forward

TGATTCTCTTGCCTCTTCTTG

Reverse

TATGGAATCTCCTGGCTCTC

TIMP-1

Forward

CTGAGAAGGGCTACCAGAG

Reverse

GTCATCGAGACCCCAAGGT

TIMP-2

Forward

GGACCTGACAAGGACATCG

Reverse

TTCTTTCCTCCAACGTCCAG

MMP-1

Forward

GATGAAAGGTGGACCAACAAT

Reverse

CCAAGAGAATGGCCGAGTTC

MMP-2

Forward

TGGGGGAGATTCTCACTTTG

Reverse

CCATCAGCGTTCCCATACTT

MMP-14

Forward

TGGGGTCATCTGCTTCTCTT

Reverse

TAGGGCTCATATGCCCAAAG

Housekeeping genes

Primers

Sequences (5'- 3')

GAPDH

Forward

CAAGTTCAACGGCACAGTCA

Reverse

CATACTCAGCACCAGCATCA

α-tubulin

Forward

CAATTCCATCCTCACCACC

Reverse

CAACCTGTTTAAGTTAGTGTAG

TBP (TATA-box binding protein)

Forward

TGCGTTGATCTTCAGTTCTG

Reverse

CTTGCTGCTAGTCTGGATTG

ACTB (β-actin)

Forward

GGTGTGATGGTGGGTATGG

Reverse

GTTGGTGACAATGCCGTGTT

**Table 3.** Quantitative Indicators of Hematological Analysis of Animals of the DOX-15, DOX-10, DOX-5 Groups
Time	Groups
DOX-15, DOX-10, DOX-5 vs. control (P < 0.05, P < 0.01, *P < 0.001). GR: granulocytes; Hb: hemoglobin; Hct: hematocrit; Lym: lymphocytes; MCH: mean concentration hemoglobin; MCHC: mean corpuscular hemoglobin concentration; MCV: mean corpuscular volume; Mi: monocytes; Plt: platelets; RBCs: red blood cells; WBCs: white blood cells.
WBCs, × 10⁹ g/L
Starting point	9.4 (7.8 - 13.7)	13.3 (7.7 - 17.3)	10.7 (7.6 - 12.7)	9.1 (8.2 - 12.9)
0 point	6.1 (4.7 - 7.4)**	10.3 (7.4 - 11.7)	7.9 (5.7 - 10.1)	12.2 (9.5 - 13.6)
First month	11.4 (7.7 - 15.5)	13.8 (13 - 14.9)	12.7 (10.3 - 12.8)	10.3 (8.5 - 13.3)
Second month	6.3 (4.1 - 7.1)	6.9 (4.9 - 11.3)	7.9 (6.3 - 9.5)*	11.2 (3.8 - 6.3)
Lym, × 10⁹ g/L
Starting point	7.1 (4.8 - 7.9)	6.2 (5 - 8.9)	8.1 (6.2 - 8.2)	7.1 (6.3 - 7.8)
0 point	3.8 (3 - 7.5)**	6.9 (5.3 - 7.9)	5.6 (4.7 - 7.8)	8.9 (7.3 - 9.8)
First month	6.8 (5.9 - 8.4)	10.5 (8.7 - 11.7)	10 (7.2 - 10.9)	7.9 (6.6 - 10.1)
Second month	3.1 (2.8 - 6)	5.7 (3.8 - 7.4)	5.6 (4.2 - 7.1)	7.6 (2.8 - 4.4)
Mi, %
Starting point	8.9 (7 - 10.3)	4.0 (0.5 - 9.9)	7.5 (0.6 - 13)	1.4 (0.6 - 8.1)
0 point	0.7 (0.6 - 2.6)	0.6 (0.6 - 0.7)**	3.1 (0.6 - 5.4)	4.4 (2.9 - 7)
First month	9 (3 - 11.4)	4.4 (1.2 - 12.3)	4.2 (3 - 12)	3.8 (3.2 - 10.4)
Second month	2.8 (1.9 - 9.5)	7.3 (3.1 - 9.7)	4.9 (2.8 - 9.8)	8.7 (6.4 - 9.8)
GR, %
Starting point	20.9 (18.3 - 24.5)	14.2 (13.4 - 21.1)	14.2 (8.5 - 18.8)	13.3 (9.2 - 18.8)
0 point	29.8 (24.1 - 35.8)	26.7 (20.6 - 31.3)	17.5 (14.5 - 20.4)	19.6 (16.3 - 23.8)
First month	19 (17.6 - 38.8)	17.9 (15.8 - 21.6)	16.4 (10.4 - 20)	15.4 (14.3 - 19.1)
Second month	12.7 (9.2 - 39.3)	16.8 (14.1 - 19.4)	22.80 (18.1 - 25.5)	24.85 (20.5 - 28.6)
RBCs, 10¹²/L
Starting point	8.3 (7.4 - 8.9)	8 (7.9 - 8.5)	9.6 (8.7 - 9.9)	9.4 (8.7 - 9.7)
0 point	8.6 (8.1 - 9.3)	9.6 (9.2 - 10.6)	8.8 (8.4 - 9.1)	8.7 (8.2 - 9.3)
First month	8.1 (7.7 - 8.5)	8.2 (6.9 - 8.8)	9.7 (9.4 - 10.6)	8.6 (8.3 - 9.6)
Second month	7.7 (6.8 - 8.8)	8 (7.6 - 8.7)	7.9 (7.7 - 8.2)	7.5 (7.2 - 7.7)
Hb, g/L
Starting point	174 (156 - 189)	166 (160 - 170.5)	168.5 (157.5 -173.8)	167 (158.5 - 170.3)
0 point	134 (128.3 -139.8)**	161 (154 - 165)	161 (155 - 164.3)	169 (162 - 172)
First month	169 (164 - 183.5)	177 (169 - 189.5)	169 (164 - 171)	168 (165.5 - 178.3)
Second month	154 (134 - 190.5)	170 (166 - 176.5)*	155.5 (152 - 160.8)	156 (150.8 - 160)
Hct, %
Starting point	43.4 (38.85 - 45.47)	40.9 (39.66 - 45.82)	46.9 (44.8 - 48.7)	44.2 (41.8 - 46.6)
0 point	42.5 (40.2 - 47.3)	48.8 (46.8 - 49.2)	40.1 (37.5 - 42.7)	42.7 (39.1 - 44.9)
First month	39.5 (38.2 - 42.2)	41.9 (39.5 - 45.4)	47.9 (45.8 - 50.4)	40.4 (38.7 - 44.9)
Second month	34.5 (29.2 - 43.7)	38.1 (36.7 - 39.9)**	35.2 (34.8 - 36.1)	41.1 (33.4 - 35.3)
MCV, fL
Starting point	52 (51 - 52)	51 (50 - 54)	47.5 (46.7 - 51.)	48 (47 - 49)
0 point	50 (50 - 50.7)	51 (48 - 51.5)	45 (45 - 45.7)	48 (46 - 50)
First month	50 (46.5 - 51.5)	50 (47.7 - 53)	48 (47 - 50)	47 (46.2 - 49.5)
Second month	46 (42.5 - 50)	47.5 (46 - 51)	44.5 (42.7 - 45)	46 (45 - 46)
MCH, pg
Starting point	21 (21 - 21.2)	20.2 (19.3 - 21.3)	17 (16 - 18.6)	17.9 (16.6 - 19.2)
0 point	15.6 (14.4 -17.4)**	16.9 (14.7 - 18.2)*	18.1 (17.6 - 18.9)	19.2 (18.3 - 19.8)
First month	21.2 (20.3 - 21.9)	21.8 (20.5 - 22.1)*	17 (16 - 17.7)	19.3 (18.4 - 19.9)
Second month	20.6 (19.4 - 21.7)	21.7 (20.3 - 22)	19.6 (19.4 - 19.4)*	20.6 (20.3 - 21.7)
MCHC, g/L
Starting point	406 (401 - 412)	406 (357 - 422)	363 (340 - 373.8)	384 (343 - 399.5)
0 point	307.5 (292.3 -350.3)*	327 (306.5 - 349.5)	392 (383 - 403)	395.5 (368 - 403.8)
First month	437 (424.5 - 443.5)	423 (414.8 - 428)	351 (340 - 357)	411.5 (364.3 - 430)
Second month	445 (435.5 - 457.5)	446.5 (437.3 - 459.3)	445.5 (439.3 -455.8)	453.5 (446.5 - 462.5)
Plt, 10⁹ g/L
Starting point	579 (566 - 649)	628 (572 - 820)	775.5 (689 - 862)	785.5 (785 - 786)
0 point	1,722 (1,358 -1,816)***	967 (852 - 1,051)	709.5 (654.5 -746.3)	644 (483 - 659.5)
First month	497.5 (409.8 - 626)	558 (510.3 - 681)	727 (593 - 820)	687 (531 - 731)
Second month	512 (437 - 643)	569 (482 - 592.8)	550 (479.5 - 567.5)	580 (564 - 616.8)

Table 3. Quantitative Indicators of Hematological Analysis of Animals of the DOX-15, DOX-10, DOX-5 Groups

Time

Groups

DOX-15

DOX-10

DOX-5

Control

*DOX-15, DOX-10, DOX-5 vs. control (*P < 0.05, **P < 0.01, ***P < 0.001). GR: granulocytes; Hb: hemoglobin; Hct: hematocrit; Lym: lymphocytes; MCH: mean concentration hemoglobin; MCHC: mean corpuscular hemoglobin concentration; MCV: mean corpuscular volume; Mi: monocytes; Plt: platelets; RBCs: red blood cells; WBCs: white blood cells.

WBCs, × 10⁹ g/L

Starting point

9.4 (7.8 - 13.7)

13.3 (7.7 - 17.3)

10.7 (7.6 - 12.7)

9.1 (8.2 - 12.9)

0 point

6.1 (4.7 - 7.4)**

10.3 (7.4 - 11.7)

7.9 (5.7 - 10.1)

12.2 (9.5 - 13.6)

First month

11.4 (7.7 - 15.5)

13.8 (13 - 14.9)

12.7 (10.3 - 12.8)

10.3 (8.5 - 13.3)

Second month

6.3 (4.1 - 7.1)

6.9 (4.9 - 11.3)

7.9 (6.3 - 9.5)*

11.2 (3.8 - 6.3)

Lym, × 10⁹ g/L

Starting point

7.1 (4.8 - 7.9)

6.2 (5 - 8.9)

8.1 (6.2 - 8.2)

7.1 (6.3 - 7.8)

0 point

3.8 (3 - 7.5)**

6.9 (5.3 - 7.9)

5.6 (4.7 - 7.8)

8.9 (7.3 - 9.8)

First month

6.8 (5.9 - 8.4)

10.5 (8.7 - 11.7)

10 (7.2 - 10.9)

7.9 (6.6 - 10.1)

Second month

3.1 (2.8 - 6)

5.7 (3.8 - 7.4)

5.6 (4.2 - 7.1)

7.6 (2.8 - 4.4)

Mi, %

Starting point

8.9 (7 - 10.3)

4.0 (0.5 - 9.9)

7.5 (0.6 - 13)

1.4 (0.6 - 8.1)

0 point

0.7 (0.6 - 2.6)

0.6 (0.6 - 0.7)**

3.1 (0.6 - 5.4)

4.4 (2.9 - 7)

First month

9 (3 - 11.4)

4.4 (1.2 - 12.3)

4.2 (3 - 12)

3.8 (3.2 - 10.4)

Second month

2.8 (1.9 - 9.5)

7.3 (3.1 - 9.7)

4.9 (2.8 - 9.8)

8.7 (6.4 - 9.8)

GR, %

Starting point

20.9 (18.3 - 24.5)

14.2 (13.4 - 21.1)

14.2 (8.5 - 18.8)

13.3 (9.2 - 18.8)

0 point

29.8 (24.1 - 35.8)

26.7 (20.6 - 31.3)

17.5 (14.5 - 20.4)

19.6 (16.3 - 23.8)

First month

19 (17.6 - 38.8)

17.9 (15.8 - 21.6)

16.4 (10.4 - 20)

15.4 (14.3 - 19.1)

Second month

12.7 (9.2 - 39.3)

16.8 (14.1 - 19.4)

22.80 (18.1 - 25.5)

24.85 (20.5 - 28.6)

RBCs, 10¹²/L

Starting point

8.3 (7.4 - 8.9)

8 (7.9 - 8.5)

9.6 (8.7 - 9.9)

9.4 (8.7 - 9.7)

0 point

8.6 (8.1 - 9.3)

9.6 (9.2 - 10.6)

8.8 (8.4 - 9.1)

8.7 (8.2 - 9.3)

First month

8.1 (7.7 - 8.5)

8.2 (6.9 - 8.8)

9.7 (9.4 - 10.6)

8.6 (8.3 - 9.6)

Second month

7.7 (6.8 - 8.8)

8 (7.6 - 8.7)

7.9 (7.7 - 8.2)

7.5 (7.2 - 7.7)

Hb, g/L

Starting point

174 (156 - 189)

166 (160 - 170.5)

168.5 (157.5 -173.8)

167 (158.5 - 170.3)

0 point

134 (128.3 -139.8)**

161 (154 - 165)

161 (155 - 164.3)

169 (162 - 172)

First month

169 (164 - 183.5)

177 (169 - 189.5)

169 (164 - 171)

168 (165.5 - 178.3)

Second month

154 (134 - 190.5)

170 (166 - 176.5)*

155.5 (152 - 160.8)

156 (150.8 - 160)

Hct, %

Starting point

43.4 (38.85 - 45.47)

40.9 (39.66 - 45.82)

46.9 (44.8 - 48.7)

44.2 (41.8 - 46.6)

0 point

42.5 (40.2 - 47.3)

48.8 (46.8 - 49.2)

40.1 (37.5 - 42.7)

42.7 (39.1 - 44.9)

First month

39.5 (38.2 - 42.2)

41.9 (39.5 - 45.4)

47.9 (45.8 - 50.4)

40.4 (38.7 - 44.9)

Second month

34.5 (29.2 - 43.7)

38.1 (36.7 - 39.9)**

35.2 (34.8 - 36.1)

41.1 (33.4 - 35.3)

MCV, fL

Starting point

52 (51 - 52)

51 (50 - 54)

47.5 (46.7 - 51.)

48 (47 - 49)

0 point

50 (50 - 50.7)

51 (48 - 51.5)

45 (45 - 45.7)

48 (46 - 50)

First month

50 (46.5 - 51.5)

50 (47.7 - 53)

48 (47 - 50)

47 (46.2 - 49.5)

Second month

46 (42.5 - 50)

47.5 (46 - 51)

44.5 (42.7 - 45)

46 (45 - 46)

MCH, pg

Starting point

21 (21 - 21.2)

20.2 (19.3 - 21.3)

17 (16 - 18.6)

17.9 (16.6 - 19.2)

0 point

15.6 (14.4 -17.4)**

16.9 (14.7 - 18.2)*

18.1 (17.6 - 18.9)

19.2 (18.3 - 19.8)

First month

21.2 (20.3 - 21.9)

21.8 (20.5 - 22.1)*

17 (16 - 17.7)

19.3 (18.4 - 19.9)

Second month

20.6 (19.4 - 21.7)

21.7 (20.3 - 22)

19.6 (19.4 - 19.4)*

20.6 (20.3 - 21.7)

MCHC, g/L

Starting point

406 (401 - 412)

406 (357 - 422)

363 (340 - 373.8)

384 (343 - 399.5)

0 point

307.5 (292.3 -350.3)*

327 (306.5 - 349.5)

392 (383 - 403)

395.5 (368 - 403.8)

First month

437 (424.5 - 443.5)

423 (414.8 - 428)

351 (340 - 357)

411.5 (364.3 - 430)

Second month

445 (435.5 - 457.5)

446.5 (437.3 - 459.3)

445.5 (439.3 -455.8)

453.5 (446.5 - 462.5)

Plt, 10⁹ g/L

Starting point

579 (566 - 649)

628 (572 - 820)

775.5 (689 - 862)

785.5 (785 - 786)

0 point

1,722 (1,358 -1,816)***

967 (852 - 1,051)

709.5 (654.5 -746.3)

644 (483 - 659.5)

First month

497.5 (409.8 - 626)

558 (510.3 - 681)

727 (593 - 820)

687 (531 - 731)

Second month

512 (437 - 643)

569 (482 - 592.8)

550 (479.5 - 567.5)

580 (564 - 616.8)