Effects of KG on Haematologic parameters in 5FU treated mice.

<p>Before and after 24 h of 5FU injection and KG treatment, about 60–100 µl of retro orbital sinus blood was collected using heparin coated capillary tube at different days (0, 3, 5 7 10 and 14) and haematological parameters such as (A) white blood cells (WBC), (B) neutrophils, (C) lymphocytes (absolute number) and (D) platelets were measured. Data were expressed as means ± SD (<i>n</i> = 8). ^*<i>P</i><0.05 ^**<i>P</i><0.01, normal Vs. 5FU and KG Vs. 5FU.</p

Invivo experimental design (A), C57BL6 (n = 40) were treated with 5FU (200 mg/kg, i.p) on day zero and post-treated (24 h) with KG (25, 50 and 100 mg/kg) for six days, daily once.

<p>Complete blood count (CBC) was taken at 0, 3, 5, 7 10 and 14 day. C57BL6 mice (n = 40) were treated with 5FU (200 mg/kg, i.p) on day zero and post-treated (24 h) with KG (25, 50 and 100 mg/kg) for six days, sacrificed on day 7 for cytokines, histopathology, gene expression and colony forming studies (B). Bone marrow cells isolated from C57BL6 were cultured and treated with KG (0.2, 2, 20 and 200 µg/ml) or Rg1 (0, 0.5, 1 and 1.5 µmol/L) at for cytokines (ELISA) and gene expression by semi quantitative and quantitative PCR (C).</p

Effects of KG on Organ weight and Histopathology.

<p>After 24 h of 5FU injection and 6 days of KG treatment, group of mice were sacrificed under ether anesthesia on day 7 and organ weights such as spleen and thymus were measured using an automated electronic balance (A) Organs such as spleen, thymus and bone marrow were removed and processed for Haematoxylin and Eosin (B). The stained tissues were examined under a light microscope (200X magnifications). Data were expressed as means ± SD. ^*<i>P</i><0.05 ^**<i>P</i><0.01, a – normal Vs. 5FU, b – KG Vs. 5FU.</p

Primers used for semi-quantitative PCR.

<p>Primers used for semi-quantitative PCR.</p

Effects of KG on genes c-Kit, FasL, IL-1, IL-2 and IL-6 in bone marrow cells.

<p>Bone marrow cells treated with KG (0.2, 2 20 and 200 µg/ml) for 4 and 8 h (n = 3). Total RNA was isolated from bone marrow cells using RNAase mini kit and analyzed by semi quantitative PCR using mouse specific primers. The relative intensities of the bands (F) were determined with the use of the ratios to β actin (normalized) and expressed as percentage (A–E).</p

Effects of KG on Haematologic parameters and body weight changes in 5FU treated mice.

<p>Before and after 24 h of 5FU injection and KG treatment, about 60–100 µl of retro orbital sinus blood was collected using heparin coated capillary tube at different days (0, 3, 5 7 10 and 14) parameters such as (A) hemoglobin, (B) red blood cells (RBC), (C) hematocrit and (D) body weight were measured. Data were expressed as means ± SD (<i>n</i> = 8). ^*<i>P</i><0.05 ^**<i>P</i><0.01, normal Vs. 5FU and KG Vs. 5FU.</p

Primers used for quantitative RT-PCR.

<p>Primers used for quantitative RT-PCR.</p

Effects of KG on genes IL-12, MCP1, TGF-β, IFN-γ and TNF-α in bone marrow cells.

<p>Bone marrow cells treated with KG (0.2, 2 20 and 200 µg/ml) for 4 and 8 h (n = 3). Total RNA was isolated from bone marrow cells using RNAase mini kit and analyzed by semi quantitative PCR using mouse specific primers. The intensities of the bands (F) were determined with the use of the ratios to β actin (normalized) and expressed as percentage (A–E).</p

Effects of KG on 5FU induced changes IL-3, GM-CSF and CFU-GM.

<p>Serum separated from blood collected from abdominal aorta of mice was subjected to cytokine analysis using Elisa kit (A). Bone marrow cells isolated from mice were grown in MethoCult methylcellulose-based complete medium in a 5% CO2 incubator for 14 d (B). Isolated bone marrow cells cultured in 6 well plated treated with KG (0.2, 2, 20 200 µg/ml) for 24 h and conditioned media was subjected to cytokine (pg/ml) analysis using commercial kit method (C &D). Data were expressed as means ± SD. ^*<i>P</i><0.05 ^**<i>P</i><0.01 compared with normal, a – normal Vs. 5FU, b – KG Vs. 5FU.</p