α­Phellandrene enhances the apoptosis of HT­29 cells induced by 5­fluorouracil by modulating the mitochondria­dependent pathway.

α­Phellandrene (α­PA), a natural constituent of herbs, inhibits cancer cell viability and proliferation. 5­Fluorouracil (5­FU) is a frequently utilized chemotherapeutic medicine for the treatment of colon cancer, which works by triggering cancer cell apoptosis. The present study examined how the combination of α­PA and 5­FU affects the suppression of human colon cancer cells by promoting apoptosis. The impact of this treatment on cell viability, apoptosis, and the expression levels of Bcl­2 family members, caspase family members and mitochondria­related molecules in HT­29 cells was assessed by the MTT assay, immunocytochemistry, western blotting and quantitative PCR. The combination of 5­FU and α­PA had a synergistic inhibitory effect on cell viability, as determined by assessing the combination index value. Bax protein expression levels were higher in the 50, 100 or 250 µM α­PA combined with 5­FU groups compared with those in the 5­FU alone group (P<0.05). By contrast, Bcl­2 protein expression levels and mitochondrial membrane potential (MMP, ΔΨm) were lower in the 100 or 250 µM α­PA combined with 5­FU groups than those in the 5­FU alone group (P<0.05). In addition, hexokinase­2 (HK­2) protein expression levels were lower in the 50, 100 or 250 µM α­PA combined with 5­FU groups than those in the 5­FU alone group (P<0.05). Compared with 5­FU alone, after HT­29 cells were treated with 50, 100 or 250 µM α­PA combined with 5­FU, the mRNA expression levels of extrinsic­induced apoptotic molecules, including caspase­8 and Bid, were higher (P<0.05). Treatment with 50, 100 or 250 µM α­PA combined with 5­FU also increased the mRNA expression levels of cytochrome c, caspase­9 and caspase­3, regulating intrinsic apoptosis (P<0.05). These results showed that α­PA and 5­FU had a synergistic effect on reducing the viability of human colon cancer HT­29 cells by inducing extrinsic and intrinsic apoptosis pathways. The mechanism by which apoptosis is induced may involve the intrinsic apoptosis pathway that activates the mitochondria­dependent pathway, including regulating the expression levels of Bcl­2 family members, including Bax, Bcl­2 and Bid, regulating MMP and HK­2 expression levels, and increasing the expression of caspase cascade molecules, including caspase­9 and caspase­3. In addition, it may involve the extrinsic apoptosis pathway that activates caspase­8 and caspase­3 leading to apoptosis.

Probiotics Fermented Bitter Melon Juice as Promising Complementary Agent for Diabetes Type 2: Study on Animal Model.

Background and Aim. Bitter melon (Momordica charantia/MC) contains charantin that has antidiabetic properties as an α-glucosidase inhibitor and antioxidative properties. Lactic acid fermentation using Lactobacillus fermentum LLB3 increased its antioxidative properties. The study was aimed to analyse the difference of the treatment that influences blood glucose and SOD level before and after treatment compared to acarbose. Experimental procedure. A total of 24 male Sprague-Dawley rats were used. Diabetes type 2 was induced by a single dose (60 mg/kg) of streptozotocin (STZ) and 120 mg/kg of nicotinamide, intraperitoneally. Following three days of STZ induction, the animals were randomly divided into four groups. Groups 1, 2, 3, and 4 were given acarbose 40 mg/100 g feed, MC 10 ml/kg body weight, fermented MC 10 ml/kg body weight, and distilled water, respectively, for 28 days. Glucose and SOD values were measured by spectrophotometer and ELISA, respectively. The difference between pretest and posttest data was analysed using the pair t-test. Data were analysed using ANOVA and Tukey HSD for post hoc analysis. Level of significance was set at 0.05. RESULTS: Fasting glucose and postprandial blood glucose were significantly decreased in groups given MC and fermented MC but not as low as those in the acarbose group (p < 0.001). The value of SOD significantly increased in groups given MC and fermented MC but not as high as those in the acarbose groups (p < 0.001). The value of SOD significantly increased in groups given MC and fermented MC but not as high as those in the acarbose groups (. CONCLUSION: Although MC gave significant results in increasing SOD and lowering fasting as well as postprandial blood glucose, fermented MC was better than MC, and acarbose still gave the best results.