Inhibitory effect of a mixture containing vitamin C, lysine, proline, epigallocatechin gallate, zinc and alpha-1-antitrypsin on lung carcinogenesis induced by benzo(a) pyrene in mice

Abstract

Background: This study was aimed to evaluate protective and therapeutic effects of a specific mixture, containing vitamin C, lysine, proline, epigallocatechin gallate and zinc, as well as alpha-1-antitrypsin protein on lung tumorigenesis induced by benzo(a) pyrene [B(a)P] in mice.

Materials and methods: Swiss albino mice were divided into two main experiments, experiment (1) the mice were injected with 100 mg/kg B(a)P and lasted for 28 weeks, while experiment (2) the mice were injected with 8 doses each of 50 mg/kg B(a)P and lasted for 16 weeks. Each experiment (1 and 2) divided into five groups, group (I) received vehicle, group (II) received the protector mixture, group (III) received the carcinogen B(a)P, group (IV) received the protector together with the carcinogen (simultaneously) and group (V) received the carcinogen then the protector (consecutively).

Results: Total sialic acid, thiobarbituric acid reactive substances, vascular epithelial growth factor, hydroxyproline levels, as well as elastase and gelatinase activities showed significant elevation in group (III) in the two experiments comparing to control group (P < 0.001). These biochemical alterations were associated with histopathological changes. Administration of the protector in group IV and group V causes significant decrease in such parameters with improvement in histopathological alterations with improvement in histopathological alterations when compared with group III in the two experiments (P < 0.001).

Conclusion: The present protector mixture has the ability to suppress neoplastic alteration and restore the biochemical and histopathological parameters towards normal on lung carcinogenesis induced by benzo(a) pyrene in mice. Furthermore, the present mixture have more protective rather than therapeutic action.

Keywords: Alpha-1-antitrypsin; benzo(a) pyrene; epigallocatechin gallate; lung tumorigenesis; lysine; proline; protector; vitamin C; zinc.

Figure 1

Effect of the protector mixture on TSA levels in serum of various studied groups of experiment (1) and (2). Data are presented as Mean ± SE, ^asignificant difference from control group (I), ^bsignificant difference from B(a) P treated group (III), *statistical significance at P < 0.001

Figure 2

Effect of the protector mixture on lipid peroxidation as expressed as TBARS in the lung tissue of various studied groups of experiment (1) and (2). Data are presented as Mean ± SE for each group, ^asignificant difference from control group (I), ^bsignificant difference from B(a)P treated group (III), *and § are statistical significance at P < 0.001 and P < 0.01 respectively

Figure 3

Effect of the protector mixture on the serum (VEGF) levels of various studied groups of experiment (1) and (2). Data are presented as Mean ± SE for each group, ^asignificant difference from control group (I), ^bsignificant difference from B(a)P treated group (III), *and # are statistical significance at *P < 0.001, ^#P < 0.05

Figure 4

Effect of the protector mixture on hydroxyproline levels in the lung tissue of various groups of experiment (1) and (2). Data are presented as Mean ± SE for each group, ^asignificant difference from control group (I), ^bsignificant difference from B(a)P treated group (III), *and § are statistical significance at P < 0.001 and P < 0.01 respectively

Figure 5

Effect of the protector mixture on elastase activity in the lung tissue of various groups of experiment (1) and (2). Data are presented as Mean ± SE for each group, ^asignificant difference from control group (I), ^bsignificant difference from B(a)P treated group (III), *statistical significance at P < 0.001

Figure 6

Gelatin zymogram showing four transparent bands in each lane corresponding to MMP-9 and MMP-2 expression, either latent or active forms in different groups

Figure 7

Represents the photomicrographs of mice lung sections in all studied groups. Sections were stained with hematoxylin and eosin (H and E, original magnification ×100). (a) Lung of control mice revealed normal architecture (Group I); (b) lung of mice treated with protector mixture alone showed no pathological changes (Group II); (c) lung of mice treated once with 100 mg B(a)P/kg (Group III₁); (d) lung of mice treated by 100 mg B(a)P/kg body weight simultaneously with protector mixture (Group IV₁); (e) lung of mice treated with 100 mg B(a)P then after 7 weeks received the protector mixture (Group V₁); (f) lung of mice treated with 8 doses each of 50 mg B(a)P (Group III₂); (g) animals were gavaged with B(a)P and treated I.P with the protector mixture after the first carcinogen dose (Group IV₁); (h) animals were gavaged with 50 mg B(a)P then treated I.P with the protector mixture after the last carcinogen dose (Group V₂)