Role of calpastatin in the regulation of mRNA expression of calpain, caspase, and heat shock protein systems in bovine muscle satellite cells.

Calpastatin participates in apoptotic cell death and cell signaling, but its role in skeletal myoblast development and molecular involvements in cell growth still remains unknown. The current study aimed to investigate the role of calpastatin on the expression patterns of calpains, caspases, and heat shock proteins (HSPs). In addition, the cell viability during myoblast growth under calpastatin silence condition was also investigated. Three small interference RNA sequences (siRNAs) were used to silence calpastatin gene and ligated into pSilencer plasmid vector to construct short hairpin RNA (shRNA) expression. The all three siRNAs significantly silence the calpastatin gene. Moreover, suppression of calpastatin significantly reduced the viability of myoblasts during growth phase when compared to control cells. Additionally, knockdown of calpastatin significantly increased the mRNA expression of µ-calpain, caspase-3, caspase-7, and caspase-9, as well as HSP-27, -70, and -90. The present study results suggested that the suppression of calpastatin resulted in the increased expression of µ-calpain, caspases, and HSPs which in turn regulate the apoptotic cell death. The present study throws light on the central role of calpastatin in the control of calpain activity, cell proliferation, cell survival, and apoptotic pathways.

Inhibitory effect of caffeic acid on cancer cell proliferation by oxidative mechanism in human HT-1080 fibrosarcoma cell line.

Caffeic acid (3,4-dihydroxy cinnamic acid) (CA) is naturally found in fruits, vegetables, olive oil, and coffee. This study was undertaken to evaluate the anticancer effect of caffeic acid on HT-1080 human fibrosarcoma cell line. The antiproliferative effect of caffeic acid was determined by MTT assay, and the oxidative stress was determined by lipid peroxidation, changes in the enzymatic, and non-enzymatic antioxidant status. To understand the mode of antiproliferative effect of CA, the authors observed intracellular ROS levels by DCFH-DA method, mitochondrial membrane potential alterations by Rh-123 staining, oxidative DNA damage by comet assay, and apoptotic morphological changes by AO/EtBr-staining method. The results show that caffeic acid enhances lipid peroxidative markers such as TBARS, CD, and LHP in HT-1080 cell line. Caffeic acid enhances the ROS levels, which is evidenced by the increased DCF fluorescence. Further, caffeic acid treatment altered the mitochondrial membrane potential in HT-1080 cells. Similarly, the authors observed increased oxidative DNA damage (% Tail DNA, % Tail length, Tail moment, and olive tail moment), and apoptotic morphological changes in caffeic acid-treated groups. These data suggest that caffeic acid exhibits potent anticancer effect in HT-1080 cell line, and that it may be used as an anticancer agent.

Antihyperlipidemic effect of bis-1,7-(2-hydroxyphenyl)-hepta-1,6-diene-3,5-dione, a curcumin analog, on nicotine and streptozotocin treated rats.

Diabetes and smoking have been considered as major health problems individually and their seriousness related to health hazard has been well reported. Data regarding the possible contribution of cigarette smoking to the development of diabetes are scarce and inconclusive. The aim was to investigate the effect of nicotine on diabetes and to analyze the effect of bis demethoxy curcumin analog (BDMCA) in streptozotocin (STZ) and nicotine-induced toxicity. The tissue lipids were extracted according to the method of Folch et al. Plasma and tissue cholesterol was estimated by the method of Allain et al. using reagent kit. Triglycerides were estimated by the method of Foster and Dunn. Free fatty acids were estimated by the method of Falholt et al. Tissue phospholipids were estimated by the method of Zilversmit and Davis. From our study, we found that nicotine not only aggravates diabetic complications but also increased the risk for diabetes. BDMCA, at a dose 80 mg/kg body weight was found to be effective in decreasing toxic effects induced by nicotine and STZ. Our data provide new evidence that cigarette smoking is an additional important factor that could be targeted for the prevention of diabetic complications.

Antitumor and antioxidant role of Chrysaora quinquecirrha (sea nettle) nematocyst venom peptide against Ehrlich ascites carcinoma in Swiss Albino mice.

This investigation aims to evaluate the antitumor and antioxidant potential of Chrysaora quinquecirrha (sea nettle) nematocyst venom on Ehrlich ascites carcinoma (EAC) tumor model. Tumor was induced in mice by intraperitoneal injection of EAC cells. The antitumor effect of sea nettle nematocyst venom (SNV) peptide was evaluated by assessing in vitro cytotoxicity, survival time, hematological, and antioxidant parameters. Intraperitoneal injection of SNV peptide increased the survival time of the EAC-bearing mice. The SNV peptide brought back the altered levels of the hematological and antioxidant parameters in a dose dependent manner in EAC-bearing mice. The results were comparable to that of the result obtained from the animals treated with the standard drug 5-fluorouracil (20 mg/kg bw). Thus, present study revealed that SNV peptide possessed significant antitumor and antioxidant activity.

Prevention of nicotine and streptozotocin treatment induced circulatory oxidative stress by bis-1,7-(2-hydroxyphenyl)-hepta-1,6-diene-3,5-dione in diabetic rats.

Diabetes and smoking have been considered as major health problems individually and their seriousness related to health hazard has been well reported. The role of nicotine in causing or worsening effect on diabetes is not well understood. The aim of our study was to investigate the effect of nicotine on experimental diabetes and to analyze the effect of bis-1,7-(2-hydroxyphenyl)-hepta-1,6-diene-3,5-dione a bisdemethoxy curcumin analog (BDMCA) in streptozotocin and nicotine induced toxicity. Group I: control rats; Group II: nicotine (2.5 mg/kg b.wt); Group III: streptozotocin (STZ) (40 mg/kg b.wt); Group IV: STZ (40 mg/kg b.wt) + nicotine (2.5 mg/kg b.wt); Group V: STZ + nicotine + BDMCA (40 mg/kg b.wt); Group VI: STZ + nicotine + BDMCA (80 mg/kg b.wt). Efficacy of BDMCA was determined by evaluating blood glucose, thiobarbituric acid reactive substances (TBARS), hydroperoxides (HP), activities of marker enzymes alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) and activities of antioxidant enzymes superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx). From our study, we have observed that nicotine not only aggravates diabetic complications but also increased the risk for diabetes. BDMCA, at a dose 80 mg/kg body weight was found to be more effective in decreasing toxic effects induced by nicotine and STZ.