Loperamide potentiates doxorubicin sensitivity in triple-negative breast cancer cells by targeting MDR1 and JNK and suppressing mTOR and Bcl-2: In vitro and molecular docking study.

Multidrug resistance (MDR) is the leading cause of treatment failure in triple-negative breast cancer (TNBC) patients treated with doxorubicin (DXR). We aimed to investigate the potential of the antidiarrheal drug Loperamide (LPR) in sensitizing TNBC cells to DXR and elucidate the underlying molecular mechanisms. Therefore, we examined the effects of DXR alone or in combination with LPR on MDA-MD-231 cells viability using MTT assay, cell cycle, and apoptosis by flow cytometry, and the expression of the MDR-related genes (MDR1 and JNK1) and cell cycle/survival genes (p21, mTOR, and Bcl-2) by quantitative reverse transcription polymerase chain reaction. Results showed that adding LPR to DXR potentiated its antiproliferation effect and reduced its IC50 by twofolds compared with DXR alone. The value of the combination index of LPR/DXR was <1 indicating a synergistic effect. Combined DXR/LPR treatment also caused G1 arrest and potentiated apoptosis more than DXR-single treatment. At the molecular levels, LPR/DXR treatment downregulated the mRNA of MDR1 (1.35-folds), JNK1 (2.5-folds), mTOR (6.6-folds), Bcl-2 (9.5-folds); while upregulated p21 gene (8-folds) compared with DXR alone. Molecular docking analyses found LPR antagonizes MDR1 and JNK1 proteins, and hence supports the in vitro studies. In conclusion, the results confirmed the potential of LPR in sensitizing TNBCs to DXR by targeting MDR1 and JNK1 and suppressing Bcl-2 and mTOR genes, while upregulating the cell cycle inhibitor gene p21. Additionally, LPR could be repurposed to reduce the therapeutic doses of DXR as indicated by the dose reduction index (DRI) and subsequently decrease its side effects.

In vitro and in vivo hypolipidemic properties of the aqueous extract of Spirulina platensis, cultivated in colored flasks under artificial illumination.

BACKGROUND: Spirulina is blue-green algae that grows mainly in tropical and subtropical lakes and is commonly used due to its nutritional features including high concentrations of protein, vitamins, mineral salts, carotenoids and antioxidants. This study aimed to investigate the anti-hypercholesterolemic potential of aqueous extract of Spirulina platensis cultivated in different colored flasks under artificial illumination; in vitro and in the diet induced hypercholesterolemic Swiss albino mice. METHODS: Spirulina platensis was cultivated in red, blue, green and colorless Erlenmeyer flasks containing Zarrouk's medium under aerobic conditions, with incessant illumination by artificial cool white fluorescent with light intensity of 2500 lux (35 µmol photon m-2 s-1). Chlorophyll a and total carotenoid contents were estimated using colorimetric methods, fatty acids composition was determined by GC-Mass, in vitro and in vivo anti-cholesterol assays were used in assessing the anti-hypercholesterolemia potential of obtained Spirulina cells. RESULTS: The results showed that the highest cell dry weight, chlorophyl a, and carotenoid of S. platensis were observed in colorless flasks and that the lowest values were recorded with the green colored flasks. Also, the hot water extract of S. platensis obtained from colorless flasks at a concentration of 15 mg/mL after 60 min of incubation exhibited the greatest reduction of cholesterol level. Gas chromatography-mass spectrometry analysis of S. platensis methanolic extract showed 15 bioactive compounds were identified and grouped according to their chemical structures. An experimental model of hypercholesterolemic mice had been examined for impact of S. platensis individually and combined with atorvastatin drug. All S. platensis groups resulted in a remarkable decrease in plasma total cholesterol, triglycerides and low density lipoprotein; and increase in high density lipoprotein. CONCLUSION: The present study concluded that the hot aqueous extract of S. platensis developed in colorless flasks is recommended as a natural source for bioactive compounds, with anti-cholesterol and antioxidant potentialities.

Lack of Any Relationship Between Circulating Autoantibodies and Interleukin­6 Levels in Egyptian Patients Infected with the Hepatitis C Virus

Introduction: Elevated serum interleukin (IL) 6 has been reported in patients infected with the hepatitis C virus (HCV), but it remains debatable whether this influences the production of autoantibodies and the biochemical profile of HCV disease. Therefore, this current study was conducted to evaluate the relationship between IL-6 and circulating autoantibody levels in HCV positive patients. Methods: Levels of IL-6 in serum samples from 102 patients with HCV and 103 normal controls were determined by enzyme linked immunosorbent assay (ELISA). Autoantibodies were detected by immunofluorescence. Results: Levels of IL-6 were significantly higher (p=0.028) in patients infected with (HCV) compared with normal group. Autoantibodies were noted in in 43.1% of the patients; of these, 23.5% featured anti-nuclear antibodies (ANA+), 16.7% anti-smooth muscle antibodies (ASMA+), 7.8% anti-mitochondrial antibodies (AMA+), 17.6% anti-parietal cell antibodies (APCA+), 7.8% anti canalicular antibodies, and 2.9% anti reticulin antibodies (ARA+). No patients were found to be positive for anti-brush border antibodies (ABBA) or anti-ribosomal antibodies. (ARiA). No links with IL-6 levels were apparent. Conclusions: IL-6 levels are increased in patients infected with HCV disease and could influence the production of autoantibodies. However, this study did not provide evidence of a specific relationship between IL6 and circulating autoantibodies in such cases.