Bradford dye was purchased from Sigma Aldrich, India, potassium dichromate, hydrogen peroxide (H2O2), glacial acetic acid was procured from Merck Pvt Ltd, India; capston cigarette from Capston, India ltd. All other chemicals were procured either from Himedia Laboratories, Mumbai, India or of analytical grade.

Bioactive compound; n-Octadecanyl-O-α-Dglucopyranosyl (6→1″)-O-α-D-glucopyranoside (F18) from FVBM extract was isolated by following the protocol Iqbal et al., 2015.

Male Sprague-Dawley (SD) rats weighed around 100- 150 gm were procured from Indian Institute of Toxicology Research Center, Lucknow. The proposed study was approved by Institutional Animal Ethics Committee (IAEC) (registration number: IU/Biotech/project/CPCSEA/13/11). The rats were housed 5 per cage for one week in the animal house for acclimatization at a temperature of 21-22°C with 12 hours light and dark cycle. The rats were given standard diet and water ad libitum.

Sequentially extracted FVBM extract, its bioactive fraction (F18) and reference drug atorvastatin were dissolved in 10 % dimethyl sulfoxide (DMSO) at different concentrations and were homogenized with saline. The doses of the extracts were selected on the basis of previously published reports Iqbal et al., 2015T S et al.,2013.

FVBM extract, its bioactive fraction (F18) and atorvastatin suspension was administered through gastric intubation in two divided doses (morning and evening) of 0.5 ml each/rat/day. Rats in smoking control group received 0.5 ml of saline containing 10 % DMSO (vehicle) twice daily while rats in normal control group received 0.5 ml of saline containing 10 % DMSO twice daily. The rats were divided randomly and equally (5 rats in each group) in groups as illustrated in Table 1 .

Rats were exposed to cigarette smoke in the morning by keeping two rats in bottomless metallic container (10 x 11 x 16 inch), having two holes of 3 and 1.5 cm diameter, one on the either side. A burning cigarette was introduced through one hole (3 cm) and the other hole (1.5 cm) was used for ventilation. Animals were exposed to CS for 30 minutes, daily for 4 weeks with interval of 10 min between each 10 min exposure, using 3 cigarettes/day/2 rats in each group Anbarasi et al., 2006.

Liver and lung were excised and kept in ice-cold saline. A portion of liver and lung were immediately fixed in 10 % neutral formalin for histopathological studies.

At the end of the experiment, liver and lung from the rats were promptly excised and chilled in ice-cold saline. After washing with saline, it was blotted and weighed. One gram of wet tissues was cut into pieces and homogenized with 9 ml of chilled 0.1 M sodium phosphate buffer, pH 7.4 (containing 1.17 % KCl) in a waring blender. The homogenate was centrifuged at 1,000 rpm for 10 min at 4°C, aliquoted and stored at -20°C. The remaining portion of the liver and lung homogenates was centrifuged at 12, 000 rpm for 20 min at 4°C. The post mitochondrial supernatant (PMS) thus obtained was also aliquoted and stored at –20°C for future use.

The enzymatic activity of catalase in PMS of liver and lung was measured by adopting the procedure of Sinha (1972). Enzymatic activity of SOD in PMS fraction of liver and lung was determined by the method as described by Kakkar et al. (1984) based on the 50 % inhibition of the formation of nicotinamide adenine dinucleotide (NADH)-phenazine methosulphatenitroblue tetrazolium formazan at 560 nm. Glutathione peroxidase activity in liver and lung homogenate was assayed by modifying of the previous protocols Hafeman et al., 1974Mills, 1959. The enzymatic activity of Gred in liver and lung was determined according to the method of Carlberg and Mannervik Eriksson et al., 1975. Method of Habig et al. (1974) was used to measure the GST activity in PMS fraction of liver and lung.

For the determination of GSH content in liver or lung homogenate, the previous methods were followed Ellman, 1959Sedlak and Lindsay, 1968.

For histopathological study, a portion of liver and lung were used. For microscopic preparation of the above tissues, method of Disbrey and Rach (1970) was used. Two formalin fixed samples from each tissue were embedded in paraffin and sectioned after block preparation. The paraffin block was sectioned in 4~5 μm thickness by using a microtome to make a slide and then it was dyed with hematoxylin-eosin (H&E) and observed under a light microscope and photographs were taken JA, 2001 .

The protein concentration of liver homogenate and PMS was analyzed by the method of Bradford (1976) using bovine serum albumin as standard. Aliquots of liver homogenates and PMS were first precipitated with 10 % TCA followed by centrifugation at 1500 rpm for 10 min. The protein pellets were dissolved in 0.5 N NaOH and used for protein determination.

For all assays, samples were analysed in triplicate and the results were expressed as mean ± SD. The results were evaluated using one-way analysis of variance (ANOVA) and two tailed Students T-test. Statistical significance were expressed as *p<0.05, **p<0.01 and ***p <0.001.

Free radicals released by cigarette smoke (CS) generate oxidative stress. This process could be responsible for alterations in the activities of enzymatic and non-enzymatic antioxidants and development of atherosclerosis. Since, protection against oxidative stress/ROS is provided by enzymatic and non-enzymatic antioxidants, therefore, the status of antioxidant enzymes, such as CAT, SOD, Gpx, GST and Gred including GSH concentrations in liver and lung of experimental smoke-exposed rats are important.

The enzymatic activities of hepatic CAT, SOD, Gred and GST in S-C rats were significantly decreased from N-C values of 62.28, 7.65, 18.29 and 132.8. U/mg protein to 32.45, 4.8, 11.28 and 68.12 U/mg protein, respectively, while Gpx activity in S-C rats was increased from N-C values of 44.2 to 57.28 U/mg protein ( Figure 1 ). The restoration in above enzymatic activities of hepatic CAT, SOD, Gred and GST in FVT-2 treated rat was 24, 32, 48 and 36 % of respective normal control values. The values of these enzymes in C-T group, in comparison to S-C values, were significantly increased by 35, 53, 60 and 43 % respectively. In contrast the hepatic Gpx activity was significantly decreased by 14, 18, 20 and 10 % in FVT- 1, FVT-2, C-T and A-T rats. As depicted in Figure 3 , the hepatic GSH content were reduced by 43 % when compared to value of N-C group. Administration of FVBM extract, F18 bioactive compound and atorvastatin to CS-exposed rats resulted in a significant increase in GSH content by 22, 32, 45 and 18 % respectively.

Moreover, similar pattern was observed in lung enzymatic and non-enzymatic antioxidant defense system. As depicted in Figure 2 the enzymatic activities of lung CAT, SOD, Gred and GST in S-C rats were significantly decreased from N-C value of 32.5, 14.7, 24.3 and 127.7 U/mg protein to 21.86, 9.56, 11.78 and 102.47 U/mg protein Figure 2 . Whereas, lung Gpx activity in SC rats was significantly increased from N-C values of 59.68 to 82.73 U/mg protein. The restoration in above enzymatic activities of lung CAT, SOD, Gred and GST in FVT-2 treated rat was 34, 40, 41 and 20 % of respective N-C values. The values of these enzymes in C-T group, in comparison to N-C values were significantly increased by 42, 50, 47 and 24 % respectively. In contrast the lung Gpx activity was significantly decreased by 11, 17, 26 and 16 % in FVT-1, FVT-2, C-T and A-T treated rats. Similarly, lung GSH content which was significantly reduced by 33 % in S-C rats as significantly increased by 20, 37, 46 and 28 % after administration of FVBM extract, F18 bioactive compound and atorvastatin Figure 3 . Here it is interesting to mention that atorvastatin also exhibited significant amelioration in all the enzymatic and non-enzymatic activities but is considered to be less effective than higher dose of FVBM extract and F18 bioactive compound.

In summary, hepatic and lung CAT, SOD, Gred, GST enzymes and non-enzymatic GSH, which constitute a mutually supportive team of defense against ROS, are significantly ameliorated after feeding of FVBM extract and F18 bioactive compound as well as substantially quenches the free radicals, thus positively normalizing the above enzyme levels close to normal values.

It is apparent from Figure 4A that histopathology of the liver of the control rats showed normal looking uniform hepatocytes with small vesicular nuclei and abundant eosinophilic granular cytoplasm with distinct cell boundaries. Architecture of liver is well maintained. Normal Interstitial cells with vascularity. CS-exposed rats showed smaller hepatocytes with small vesicular or pyknotic nuclei and reduced granularity of cytoplasm. Cytoplasmic boundaries are relatively indistinct. Architecture not well maintained. Interstitial cells increased and vascularity is reduced ( Figure 4B ). Administration of FVBM extract at lower dose has shown smaller hepatocytes with smaller vesicular nuclei and eosinophilic cytoplasm with indistinct cell boundaries showing proliferation. Architecture not well maintained but vascularity has relatively increased ( Figure 4C ). Moreover, liver section of FVT-2 rats (higher dose) ( Figure 4D ) showed proliferated normal hepatocytes with normal vesicular nuclei and abundant eosinophilic cytoplasm with distinct cell boundaries. Architecture well maintained with normal interstitial cell and vascularity. Similarly, liver section of CT rats showed proliferative hepatocytes with normal morphology as well as architecture. Interstitial cells normal with normal vascularity, also no toxic effects noted ( Figure 4E ). Furthermore, liver section of A-T rats showed normal hepatocytes with normal vesicular nuclei and abundant eosinophilic cytoplasm alongwith well maintained normal architecture and vascularity ( Figure 4F ).

Histopathology of lung in N-C group shows normal lung stroma and characteristic spongy appearance of the lung with normal looking numerous alveolar spaces, had normal blood vessels and bronchi. The bronchoalveolar unit parenchyma in the normal lung was within the limits ( Figure 5A ). The lung section of CSexposed rats showed the increased volume of stroma with presence of few collections of stromal cells with reduced alveolar spaces, reduced vascularity and constricted bronchi ( Figure 5B ). There was obliteration of most alveoli and subsequently compensatory dilatation and expansion of the contiguous ones with destruction of alveolar wall. On treatment with lower dose of FVBM extract, section of lung shows reduced stroma, increased alveolar spaces, vascularity with patent blood vessels and dilated bronchi ( Figure 5C ). On the other hand, treatment with higher dose, section of lung showed changes as described above but bronchi are relatively constricted ( Figure 5D ). In addition, in lung section of C-T rat observed that stroma is further reduced with presence of large alveolar spaces. Blood vessels were normal but smaller bronchi were further constricted ( Figure 5E ). Furthermore, lung slice of atorvastatin treated group showed the alveolar spaces comparatively less than normal with corresponding increase in stromal elements. Blood vessels and bronchi were normal in the presence of few constricted smaller bronchi ( Figure 5F ).