Anti-snake venom activities of extracts and fractions from callus cultures of Sapindus saponaria

Figures & data

Figure 1.  Inhibition of the PLA₂ activity of snake venoms (A and B) and isolated toxins (C and D) induced by the extracts and fractions of S. saponaria, preincubated at different ratios (w/w, venoms/proteins: Extracts/fractions) for 30 min at 37°C. Each experiment represents the mean ± S.D. (n = 6).

Figure 2.  Inhibition of the edema-inducing activity of crude snake venom (A) and isolated toxins (B and C) after incubation for 30 min at 37°C with extracts (B and C) or fractions of S. saponaria (A1, A2, B2) at ratios of 1:50 and 1:10 (w/w) to venom and toxins, respectively. Each denoted value represents the mean ± S.D. (n = 6).

Figure 3.  Inhibition of the hemorrhagic activity of crude snake venoms by the extracts and fractions of S. saponaria preincubated at different ratios for 30 min at 37°C. Each denoted value represents the mean ± S.D. (n = 6).

Figure 4.  Inhibition of the myotoxic activity of B. jararacussu venom and BthTX-I toxin after incubation for 30 min at 37°C with fractions of S. saponaria at a ratio of 1:50 (w/w, venom or toxin/fractions). Each denoted value represents the mean ± S.D. (n = 6).

Table 1.  Neutralization of the clotting activity induced by B. jararacussu and C. d. terrificus crude venoms preincubated with extracts and fractions of S. saponaria.

Samples	Time of coagulation (sec)#
	Without extracts or fractions	1:5 (w/w)
PBS + Ca^2+	310 ± 0.12	-
B. jussu (50 µg)	60 ± 0.07	-
A1	-	188 ± 0.02
A2	-	155 ± 0.06
B2	-	75 ± 0.12
Extract B	-	120 ± 0.08
Extract C	-	109 ± 0.20
C. d. terrificus (50 µg)	60 ± 0.10	-
A1	-	63 ± 0.07
A2	-	120 ± 0.17
B2	-	80 ± 0.08
Extract B	-	67 ± 0.03
Extract C	-	69 ± 0.05

^#Each experiment is represented as mean ± S.D. (n = 6).

Table 2.  Survival time of mice injected with C. d. terrificus venom and crotoxin preincubated with extracts and fractions of S. saponaria.

Samples	Survival time (min)*
	Without extracts or fractions	1:10 (w/w)	1:50 (w/w)
C. d. terrificus (20 µg)	65 ± 0.02	–	–
A1	–	195 ± 0.22	201 ± 0.18
A2	–	136 ± 0.20	142 ± 0.15
B2	–	158 ± 0.09	158 ± 0.04
Extract B	–	164 ± 0.14	174 ± 0.11
Extract C	–	74 ± 0.14	85 ± 0.03
Crotoxin (10 µg)	72 ± 0.09	–	–
A1	–	186 ± 0.18	190 ± 0.13
A2	–	165 ± 0.10	175 ± 0.19
B2	–	148 ± 0.05	149 ± 0.07
Extract B	–	108 ± 0.12	123 ± 0.12
Extract C	–	92 ± 0.05	92 ± 0.10
Extracts and fractions (500 µg)	#	–	–

^#100% of animal survival (>48 h)

*Each experiment is represented as mean ± S.D. (n = 8).

Figure 5.  TLC profiles of extracts and fractions from S. saponaria callus culture and standards. Mobile phase: (A): Hex:EtOAc (7:3); (B): CHCl₃:MeOH (8:2); (C): BAW (n-butanol:acetic acid:water, 4:1:5, upper phase). Color reagents: (A) and (B): Vanillin-sulfuric acid; (C): Ninhydrin. Extracts and fractions are represented by their respective nominations (B, C, A1, A2, B1, B2, N1 and N2). Also shown are the standards of β-sitosterol (SI), stigmasterol (ST) and β-sitosterol glucoside (SG).

Figure 6.  Effect of stigmasterol (A) on myotoxic (B and C) activity induced by isolated myotoxins, BthTX-I and BthTX-II, respectively. Each experiment is represented as mean ± S.D. (n = 6).