The CYP3A4derived 57 bp fragment comprising a consensus YY1-binding site inserted into the CYP3A5 promoter inhibited its transcriptional activity in renal cells

nohistochemistry for Caspase-3 was performed for identification of apoptotic cells using a combination of streptovidin-biotin-peroxidase method and microwave antigen retrieval on TGF-b2 Reduces MTX Induced Intestinal Injury containing equal amounts of total protein were resolved by SDS-PAGE under reducing conditions. After electrophoresis, proteins were transferred to a PVDF membrane and probed with various primary antibody to anti-bcl-2 antibody, anti-bax antibody, anti-phosphoERK antibody, anti-b-catenin antibody, anti-TGF-b2 receptor antibody, anti-ERK2 antibody, anti-IL-1B and anti-b-actin antibody which were purchased from Santa Cruz Biotechnology. Horseradish peroxidaseconjugated secondary antibody was purchased from Jackson ImmunoResearch Laboratories Inc and an enhanced chemiluminescent substrate from Biological Industries. The optical density of the specific protein bands was quantified by using a densitometer. Expression of bax and bcl-2 genes Expression of bax and bcl-2 levels was determined by quantitative real-time PCR on cDNA samples using Cyber Green Master Mix with the PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/22212322 exception of template and primers. Primers for LY2109761 chemical information Rattus norvegicus bax and bcl-2 were synthesized by Syntezza Bioscience ltd. Israel, and 18 s rRNA Control kit was purchased from Eurogentec, EGT Group. compared to control animals . MTX rats demonstrated a significant decrease in final body weight compared to control animals. Although MTX-TGF-b rats demonstrated a trend toward an increase in final body weight compared to MTX animals, this trend did not achieve statistical significance. Eighty percent of MTX rats showed mild to moderate diarrhea. Treatment with TGF-b2 did not change stool patterns compared to MTX-non treated animals. Intestinal mucosal parameters. Treatment with TGF-b2 led to a significant increase in jejunal bowel weight, as well as in jejunal and ileal mucosal weight compared to control-untreated animals . Treatment with MTX resulted in a significant decrease in bowel weight in jejunum and ileum, mucosal weight in jejunum and ileum, mucosal DNA in jejunum and ileum , and mucosal protein in jejunum and ileum compared to control animals. Treatment of MTX rats with TGF-b2 led to a significant increase in jejunal and ileal bowel weight, jejunal and ileal mucosal weight, jejunal and ileal mucosal DNA content, and jejunal and ileal mucosal protein content when compared to MTX-animals. Statistical analysis The data are expressed as the mean 6 SEM. A one-way ANOVA for comparison, followed by Tukey’s test for pair-wise comparison was used for statistical analysis. Prism software was used and statistical significance was defined as P,0.05. Intestinal histopathology Treatment of control rats with TGF-b2 did not change significantly Park’s score, villus height and crypt depth in jejunum and ileum compared to control non-treated animals. Microscopic analysis of the intestine 72 hours after MTX injection revealed a characteristic change of intestinal damage, including a significant epithelial atrophy, blunting of the villi and signs of crypt remodeling which was accompanied by marked cellularity, mainly with mononuclear cells in the lamina propria, the presence of flattened and vacuolated cells, and an increased number of blood vessels in the stroma. Consistent with these findings, the intestinal injury score increased significantly in MTX rats in both jejunum and ileum compared to control rats. Following TGF-b2 administration, MTX rats showed less signif

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