Protective effects of some Nigerian indigenous antimalarial plants on placental malaria related pathological damages and pregnancy outcomes in murine model

Figures & data

Table 1. Experimental groups

Groups	Pregnancy status	Infection status	Treatment status	No of groups		Total Groups
				Treatment	Dosage
1	Pregnant	Infected	Plants (3 plants)	3	3	9
2	Pregnant	Infected	Treatment(SP)	1	1	1
3	Pregnant	Infected	No treatment	1	1	1
4	Pregnant	Not infected	No treatment	1	1	1
5	Pregnant	Not infected	Plants (3 plants)	3	3	9
6	Pregnant	Not infected	Treatment (SP)	1	1	1
Total						22

Note: 3 plants (M. lucida, C. citratus, and E. chlorantha) extracts were evaluated in this study at 3 different dosages (100, 200 and 300 mg/kg body weight). SP = Sulphadoxine pyrimethamine (at a single dose of 10 mg/kg)

Table 2. Body weight changes of infected and non-infected pregnant mice treated with the plant extracts compared with the control at gestation days (GD) 1, 9, and 17

Treatment/mg/kg	Infected Pregnant				Non-Infected Pregnant
	GD 1	GD 9	GD 17		GD 1	GD 9	GD 17
E. chlorantha 100	22.40 ± 0.25^a	24.40 ± 0.51^a	22.60 ± 0.60^a		21.80 ± 0.37^a	24.20 ± 0.37^a	22.40 ± 0.51^a
E. chlorantha 200	23.20 ± 0.20^a	26.00 ± 0.55^a	24.80 ± 0.37^a		23.20 ± 0.20^a	26.00 ± 0.55^a	24.60 ± 0.40^a
E. chlorantha 300	22.80 ± 0.37^a	25.60 ± 0.68^a	23.60 ± 0.50^a		22.60 ± 0.51^a	25.40 ± 0.75^a	23.20 ± 0.58^a
C. citatrus 100	22.80 ± 0.37^a	26.40 ± 0.51^a	35.00 ± 0.55^b		22.80 ± 0.37^a	26.00 ± 0.55^a	34.60 ± 0.60^b
C. citatrus 200	22.60 ± 0.40^a	25.40 ± 0.60^a	33.00 ± 0.32^b		22.20 ± 0.20^a	25.00 ± 0.45^a	32.80 ± 0.37^b
C. citatrus 300	21.40 ± 0.51^a	24.60 ± 0.87^a	31.00 ± 0.55^b		21.40 ± 0.51^a	25.00 ± 0.84^a	31.20 ± 0.66^b
M. lucida 100	22.00 ± 0.55^a	25.4 ± 0.68^a	31.20 ± 0.49^b		22.20 ± 0.37^a	25.60 ± 0.68^a	31.40 ± 0.40^b
M. lucida 200	22.60 ± 0.68^a	25.40 ± 0.51^a	34.2 ± 1.59^b		22.80 ± 0.37^a	25.80 ± 0.37^a	34.80 ± 1.39^b
M. lucida 300	22.40 ± 0.35^a	24.60 ± 0.60^a	35.00 ± 0.32^b		22.40 ± 0.68^a	24.60 ± 0.60^a	35.40 ± 0.40^b
Fansidar	21.80 ± 0.23^a	24.2 ± 0.66^a	34.00 ± 0.20^b		22.00 ± 0.32^a	24.40 ± 0.51^a	36.10 ± 0.20^b
Untreated	22.60 ± 0.13^a	26.00 ± 0.32^a	34.20 ± 0.58^b		22.60 ± 0.25^a	26.20 ± 0.37^a	35.00 ± 0.45^b
P-Value	0.176	0.191	0.000*		0.132	0.175	0.000*

Note: P-Values with asterisk (*) Connotes significant difference. Values were presented as mean ± standard error of mean (S.E.M)

Table 3. Daily chemosupression in treated and control groups of pregnant-infected mice

Treatment Groups Mg/Kg	Day1 (%)	Day2 (%)	Day3 (%)	Day4 (%)	Day5 (%)	Day6 (%)
E. chlorantha 100	2.94 ± 0.01 ^c	7.35 ± 1.15 ^c	14.71 ± 0.69 ^c	38.24 ± 1.15 ^c	55.88 ± 0.33^d	58.82 ± 3.45 ^c
E. chlorantha 200	5.08 ± 1.15^d	10.17 ± 0.01^d	22.03 ± 1.15^e	49.15 ± 0.69^e	59.32 ± 1.21^e	66.10 ± 0.33^d
E. chlorantha 300	10.34 ± 0.01^e	24.24 ± 1.10 ^f	25.25 ± 0.70 ^f	50.02 ± 1.15^e	68.19 ± 0.33 ^g	74.32 ± 0.57^ef
C. citratus 100	0.17 ± 0.17^b	3.45 ± 0.01^b	10.34 ± 1.15^b	31.03 ± 0.33^b	44.83 ± 0.58 ^c	44.83 ± 2.89^b
C. citratus 200	1.92 ± 0.03^bc	3.85 ± 0.01^b	15.38 ± 0.70 ^c	30.77 ± 1.15^b	42.31 ± 0.33^b	46.15 ± 0.76^b
C. citratus 300	1.85 ± 0.01^bc	11.11 ± 0.06^d	18.52 ± 0.30^d	37.04 ± 0.02 ^c	44.44 ± 0.25 ^c	55.56 ± 0.76 ^c
M. lucida 100	10.34 ± 0.01^e	20.69 ± 0.40^e	29.31 ± 1.18 ^g	46.55 ± 0.58^d	58.62 ± 0.36^e	67.24 ± 0.24^d
M. lucida 200	10.42 ± 0.03^e	29.17 ± 1.15 ^g	35.42 ± 0.01 ^h	56.25 ± 1.15 ^f	64.58 ± 0.69 ^f	70.83 ± 0.48^de
M. lucida 300	16.05 ± 1.15 ^f	30.86 ± 0.01 ^g	50.62 ± 1.16^1	67.90 ± 0.70 ^g	70.37 ± 0.69 ^h	77.26 ± 0.69 ^f
Fansidar	11.37 ± 0.02^e	38.57 ± 0.10 ^h	66.09 ± 1.15 ^j	98.43 ± 0.69 ^h	100.00 ± 0.00^i	100.00 ± 00 ^g

Note: Values with different superscript in a column are statistically different (p < 0.05). Values are presented as mean±Standard Error of Mean (S.E.M)

Figure 1.: Effect of M. lucida and C. citratus extracts on placenta parasitemia

Note: Placenta parasitemia for E. chlorantha could not be computed as all mice treated with E. chlorantha had abortion (fetal resorption). Histology of the oviduct of mice treated with E. chlorantha showed well-developed endometrial glands which is an indication of prevented pregnancy (Figure 2)

Figure 2. Section of (a) Oviduct of mice treated with E. chlorantha extract: showing well-formed glands (long arrows) in the endometrial submucosa. The epithelia lining is normal (short arrows), H and E × 400 (b) Uterus of mice treated with E. chlorantha extract showing fetal resorption (long arrows) Mag ×1.5

Figure 3. (a-d): Photomicrograph of cross histological section of the placenta from mice showing; A (infected not treated), B (not infected not treated) C (infected treated with fansidar) D (not infected treated with fansidar) (a) Trophoblastic cells that appear pyknotic (long arrow) in the infected mouse, degeneration and coagulative necrosis (small arrow) of the labyrinth, parasitized red blood cells and malaria parasite pigments). (b-d) No visible lesions in both infected (treated) and non-infected groups. H and E × 400

NB: PRBC connotes Parasitized red blood cells while MP means Malaria parasite

Figure 4. Photomicrograph of cross histological section of the placenta from mice treated with different concentrations of C. citratus; (a-c) Infected mice treated with 100, 200 and 300 mg/kg body weight respectively. (d-f) Non-infected mice treated with 100, 200 and 300 mg/kg body weight, respectively. (a-c) few trophoblastic cells that appear pyknotic in the infected-treated group (Long arrow). (d-f) No visible lesions in non-infected groups. Parasitized red blood cells (PRBC). H and E × 400

NB: PRBC connotes Parasitized red blood cells while MP means Malaria parasite

Figure 5. (a-f): Photomicrograph of cross histological section of the placenta from mice treated with different concentrations of M. lucida; (a-c) Infected mice treated with 100, 200 and 300 mg/kg body weight respectively. (d-f) Non-infected mice treated with 100, 200, and 300 mg/kg body weight, respectively. (a) Few trophoblastic cells that appear pyknotic in the infected-treated group (Long arrow). (b-f) No visible lesions in both infected and non-infected group, H and E × 400

Table 4. Effect of plant extracts on pregnancy outcome of infected and non-infected pregnant mice

Groups	Pups Weight (g)		Litter size		Pups mortality (%)		Litter day
	Infected	Non-infected	Infected	Non-infected	Infected	Non-infected	Infected	Non-infected
C. citratus 100	2.23 ± 0.09^a	2.33 ± 0.09^a	6.00 ± 0.00^a	7.33 ± 0.67^b	0.00^a	0.00^a	22.00 ± 0.00^b	21.67 ± 0.33^ab
C. citratus 200	1.23 ± 0.08^a	1.33 ± 0.09^a	4.00 ± 0.58^a	5.33 ± 0.33^b	25.00^b	0.00^a	22.33 ± 0.33^b	21.67 ± 0.33^ab
C. citratus 300	-	1.57 ± 0.15	-	4.67 ± 0.33	-	0.00	-	21.33 ± 0.33
M. lucida 100	1.90 ± 0.10^a	2.17 ± 0.07^b	4.67 ± 0.88^a	6.67 ± 0.88^b	0.00^a	0.00^a	21.67 ± 0.31^a	21.67 ± 0.37^a
M. lucida 200	1.97 ± 0.43^a	2.27 ± 0.23^b	4.00 ± 0.27^a	6.67 ± 1.20^b	8.25^b	0.00^a	22.00 ± 0.00^a	22.00 ± 0.00^a
M. lucida 300	-	2.30 ± 0.25	-	5.33 ± 0.88	-	0.00	-	21.33 ± 0.33
Fansidar	2.53 ± 0.34^a	2.63 ± 0.34^a	11.33 ± 0.88^a	11.67 ± 0.88^a	0.00^a	0.00^a	21.00 ± 0.00^a	20.00 ± 0.00^a
Untreated	2.63 ± 0.32^a	3.03 ± 0.12^b	5.33 ± 0.67^a	11.67 ± 0.88^b	0.00^a	0.00^a	20.33 ± 0.33^a	21.00 ± 0.00^a

Note: Values with different superscript in a row are statistically different (p < 0.05). Values are presented as mean ± Standard Error of Mean (S.E.M). All cells without values means mortality of all pregnant mice (in the group) due to miscarriage and rupture of the uteri (Figure 6).

Figure 6. Uterus of mice showing Miscarriage and Necrosis of the mice’s uterus