Analysis of the therapeutic role of platelet-rich plasma against cisplatin-induced hepatotoxicity in rats: controversy between oxidative and apoptotic markers

Cisplatin is a potent chemotherapeutic agent used to treat a variety of cancers such as ovarian, uterine, and bladder. The major limiting side effect of cisplatin is its hepatotoxicity. The possible mechanism of cisplatin hepatotoxicity was due to the affection of oxidant-antioxidant system. Platelet-rich plasma (PRP) has a powerful therapeutic option for its ability to deliver a great variety of biologically active GFs to the site of injury. PRP has grown as an attractive biologic instrument in regenerative medicine for its powerful healing properties. It is considered as a source of growth factors that may induce tissue repairing and improve fibrosis. This product has proven its efficacy in multiple studies, but its effect on cisplatin-induced hepatotoxicity has not yet been elucidated. The present study was designed to analyze the therapeutic role of PRP in cisplatin-induced hepatotoxicity. 30 adult male adult male albino rats were used in the present study divided into 3 groups (control group, cisplatin-treated group and PRP-treated group). By the end of the experimental period, blood samples were collected for measurement of serum AST, ALT and ALP enzymes; then the rats were sacrificed by cervical dislocation. Fresh liver parts were used to measure the oxidative markers in liver homogenates, while other parts were processed and subjected for histopathological and histomorphometric and immunohistochemical analyses for VEGF, Caspase 3 expression of the different experimental groups. The statistical study was done for the resultant data. Group II (Cisplatin-treated group) showed marked pathological hepatic changes; loss of architecture, congested dilated sinusoids lined by darkly stained pyknotic Kupffer cells; and hepatocytes nuclei were pyknotic and karyolitic. Dilated congested portal vein, interstitial acidophilic exudate, marked polymorphic cellular infiltration. There were increased collagen fibers deposition, a weak positive PAS reaction, strong positive caspase 3 reactions and strong positive VEGF reaction. Also, there were a marked increase in hepatic enzymes, MDA levels and a marked decrease in GSH level. Treatment with PRP in Group III revealed improvement of the hepatic parenchymal architecture with strong PAS reaction and minimal collagen fibers deposition. Weak positive caspase immunoreaction and strong positive VEGF reaction were noticed. Also, there was a marked improvement in the parameter of hepatic enzymes, MDA and GSH level comparable with the control group. It is concluded that PRP could ameliorate the liver against cisplatin-induced hepatotoxicity

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The mixed human umbilical cord blood-derived mesenchymal stem cells show higher antitumor effect against C6 cells than the single in vitro.

OBJECTIVE: It is difficult for the treatment of neurologic tumors with current therapies, including glioma. Despite the advances in cancer therapeutics, the outcomes in these patients remain poor and, therefore, new modalities are required. Recent findings have demonstrated that human umbilical cord blood mesenchymal stem cells (UCB-MSCs) have the potential to inhibit glioma cell growth in vitro. Caspase-3 plays a critical role as an executioner of apoptosis and the level of caspase-3 expression determines the degree of apoptosis in cancer cell lines. CD133 (+) glioma displays a strong resistance to chemotherapy. Interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) are important cytokines in the generation of antitumor immunity. METHODS: UCB-MSCs were harvested by density gradient separation. Green fluorescence protein stable C6 cells were established. Cytotoxicity was detected by visual survival cell assay, and caspase-3 activity was assessed using immunohistochemistry staining and western blot. Flow cytometry was used to test CD133 positive C6 cells. The concentrations of IL-2 and IFN-gamma proteins secreted from UCB-MSCs were then quantified using enzyme-linked immunosorbent assay. The cytotoxicities of IL-2 alone, IFN-gamma alone, and combination of IL-2 and IFN-gamma were compared against malignant glioma cells. RESULTS: We noted a significant cytotoxicity of UCB-MSCs for malignant glioma cells. In addition, the toxicity of mixed UCB-MSCs was significantly higher than that of single UCB-MSCs for C6 glioma cells. Capase-3 levels in UCB-MSCs treatment at an effector/target (E/T) ratio of (5+5):1 were higher than those at an E/T ratio of 10:1. On the contrary, there was no change in the protein expression of capase-3 at an E/T ratio of 0:1. Immunohistochemistry staining and western blot revealed that the expression of capase-3 was higher in mixed UCB-MSCs treatment, when compared with single UCB-MSCs. We identified reductions of approximately 39 and 73% in the number of CD133 positive C6 cells treated with the single (10:1) and the mixed [(5+5):1] UCB-MSCs respectively as compared to the control group (0:1) in transwell inserts. However, the mixed UCB-MSCs secreted more immune response-related proteins (IL-2 and IFN-gamma) than the single UCB-MSCs. Combination of IL-2 and IFN-gamma might prove more cytotoxic on target cells than IL-2 alone and IFN-gamma alone. DISCUSSION: The data collected herein confirm for the first time that the mixed UCB-MSCs were shown to have more cytotoxic effects than the single UCB-MSCs through increasing the expression of caspase-3 and decreasing the expression of CD133 in C6 glioma cells. In addition, the mixed UCB-MSCs secrete more immune response-related proteins (IL-2 and IFN-gamma) than the single UCB-MSCs. Combination of IL-2 and IFN-gamma was shown to have more cytotoxic effects than IL-2 alone and IFN-gamma alone. These results demonstrate that the mixed UCB-MSCs are a potential new therapeutic agent for glioma.

Lung endothelial monocyte-activating protein 2 is a mediator of cigarette smoke-induced emphysema in mice.

Pulmonary emphysema is a disease characterized by alveolar cellular loss and inflammation. Recently, excessive apoptosis of structural alveolar cells has emerged as a major mechanism in the development of emphysema. Here, we investigated the proapoptotic and monocyte chemoattractant cytokine endothelial monocyte-activating protein 2 (EMAPII). Lung-specific overexpression of EMAPII in mice caused simplification of alveolar structures, apoptosis, and macrophage accumulation, compared with that in control transgenic mice. Additionally, in a mouse model of cigarette smoke-induced (CS-induced) emphysema, EMAPII levels were significantly increased in murine lungs. This upregulation was necessary for emphysema development, as neutralizing antibodies to EMAPII resulted in reduced alveolar cell apoptosis, inflammation, and emphysema-associated structural changes in alveoli and small airways and improved lung function. The mechanism of EMAPII upregulation involved an apoptosis-dependent feed-forward loop, since caspase-3 instillation in the lung markedly increased EMAPII expression, while caspase inhibition decreased its production, even in transgenic EMAPII mice. These findings may have clinical significance, as both current smokers and ex-smoker chronic obstructive pulmonary disease (COPD) patients had increased levels of secreted EMAPII in the bronchoalveolar lavage fluid compared with that of nonsmokers. In conclusion, we suggest that EMAPII perpetuates the mechanism of CS-induced lung emphysema in mice and, given its secretory nature, is a suitable target for neutralization antibody therapy.