title: MAK-4 and -5 supplemented diet inhibits liver carcinogenesis in mice.
abstract: BACKGROUND
  Maharishi Amrit Kalash (MAK) is an herbal formulation composed of two herbal mixtures, MAK-4 and MAK-5. These preparations are part of a natural health care system from India, known as Maharishi Ayur-Veda. MAK-4 and MAK-5 are each composed of different herbs and are said to have maximum benefit when used in combination. This investigation evaluated the cancer inhibiting effects of MAK-4 and MAK-5, in vitro and in vivo.
  
  
  METHODS
  In vitro assays: Aqueous extracts of MAK-4 and MAK-5 were tested for effects on ras induced cell transformation in the Rat 6 cell line assessed by focus formation assay. In vivo assays: Urethane-treated mice were put on a standard pellet diet or a diet supplemented with MAK-4, MAK-5 or both. At 36 weeks, livers were examined for tumors, sera for oxygen radical absorbance capacity (ORAC), and liver homogenates for enzyme activities of glutathione peroxidase (GPX), glutathione-S-transferase (GST), and NAD(P)H: quinone reductase (QR). Liver fragments of MAK-fed mice were analyzed for connexin (cx) protein expression.
  
  
  RESULTS
  MAK-5 and a combination of MAK-5 plus MAK-4, inhibited ras-induced cell transformation. In MAK-4, MAK-5 and MAK4+5-treated mice we observed a 35%, 27% and 46% reduction in the development of urethane-induced liver nodules respectively. MAK-4 and MAK4+5-treated mice had a significantly higher ORAC value (P < 0.05) compared to controls (200.2 +/- 33.7 and 191.6 +/- 32.2 vs. 152.2 +/- 15.7 ORAC units, respectively). The urethane-treated MAK-4, MAK-5 and MAK4+5-fed mice had significantly higher activities of liver cytosolic enzymes compared to the urethane-treated controls and to untreated mice: GPX(0.23 +/- 0.08, 0.21 +/- 0.05, 0.25 +/- 0.04, 0.20 +/- 0.05, 0.21 +/- 0.03 U/mg protein, respectively), GST (2.0 +/- 0.4, 2.0 +/- 0.6, 2.1 +/- 0.3, 1.7 +/- 0.2, 1.7 +/- 0.2 U/mg protein, respectively) and QR (0.13 +/- 0.02, 0.12 +/- 0.06, 0.15 +/- 0.03, 0.1 +/- 0.04, 0.11 +/- 0.03 U/mg protein, respectively). Livers of MAK-treated mice showed a time-dependent increased expression of cx32.
  
  
  CONCLUSIONS
  Our results show that a MAK-supplemented diet inhibits liver carcinogenesis in urethane-treated mice. The prevention of excessive oxidative damage and the up-regulation of connexin expression are two of the possible effects of these products.
authors.affiliation: Laboratory of Biotechnology, Civic Hospital of Brescia, Brescia, Italy. marialetiziapenza@inwind.it <marialetiziapenza@inwind.it>
authors.forename: Marialetizia
authors.initials: M
authors.lastname: Penza
authors.forename: Claudia
authors.initials: C
authors.lastname: Montani
authors.forename: Marija
authors.initials: M
authors.lastname: Jeremic
authors.forename: Giovanna
authors.initials: G
authors.lastname: Mazzoleni
authors.forename: W L Wendy
authors.initials: WL
authors.lastname: Hsiao
authors.forename: Maurizio
authors.initials: M
authors.lastname: Marra
authors.forename: Hari
authors.initials: H
authors.lastname: Sharma
authors.forename: Diego
authors.initials: D
authors.lastname: Di Lorenzo
mesh_terms: D000818:Animals; D016588:Anticarcinogenic Agents / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D000975:Antioxidants / Q000378:metabolism*; D004195:Disease Models, Animal; D005979:Glutathione Peroxidase / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D066298:In Vitro Techniques; D008099:Liver / Q000187:drug effects* / Q000201:enzymology*; D008515:Medicine, Ayurvedic; D051379:Mice; D008862:Microsomes, Liver / Q000201:enzymology / Q000378:metabolism; D010084:Oxidation-Reduction; D028321:Plant Preparations / Q000494:pharmacology*; D011808:Quinone Reductases / Q000378:metabolism; D051381:Rats; D014520:Urethane
chemical_list: D016588:Anticarcinogenic Agents; D000970:Antineoplastic Agents; D000975:Antioxidants; C476033:Maharishi Amrit Kalash; D028321:Plant Preparations; D014520:Urethane; D005979:Glutathione Peroxidase; C041831:NADH dehydrogenase (quinone); D011808:Quinone Reductases; D005982:Glutathione Transferase