Publication: Effects Of Paddy Waste Products In Prostate Disease Using In Vitro And In Vivo Models
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Date
2018-06
Authors
Murtey, Mogana Das
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Abstract
Benign prostate hyperplasia (BPH) is common in elderly men and prostate cancer is the second leading cause of death among men. Previously, paddy waste (both husk and straw) reportedly have chemopreventive potential. The main aim of this study was to explore the chemopreventive property of paddy waste against prostate disease. This study determines the inhibitory and anti-proliferative activity of the paddy waste product in androgen-independent prostate cancer cell line, DU145 and spontaneously hypertensive rat (SHR) respectively; the effects of paddy waste products on proliferation and apoptosis related genes and protein expression in DU145; the effect of aqueous methanol extracts on histology, PCNA and androgen receptor expressions in SHR.There were two types solvent used for paddy husk and straw extracts in this study; water and aqueous methanol. Cell culture experiments revealed that the aqueous methanol extracts have better inhibitory effect than water extracts on DU145 without toxicity. Apoptosis analysis revealed that aqueous methanol extracts exhibit apoptosis inducing effect on DU145 and this claim was supported with DNA analysis (presence of bands which indicate DNA degradation) and microscopy analysis (cell shrinkage, membrane blebbing and apoptotic bodies). Western blot analysis reveals that apoptosis induced through caspase 3-mediated intrinsic pathway. Two pro-apoptotic proteins, Bax and Bad expressed higher (p<0.05) and anti-apoptotic protein, Bcl-2 downregulated (p<0.01) after treatment. This leads to increase (p<O.O 1) of caspase 9 expression which in turn activate caspase 3 and 7 (p<0.01) which lead cells to apoptosis. Aqueous methanol extracts were chosen over water extracts for the in vivo study on the SHR model. SHRs were given drinking water mixed with aqueous methanol extracts for 17 weeks showed no toxicity effect on dietary intake, body weight and vital organ (liver and kidney). It managed to reduce blood pressure [72.0mmHg (p<0.01)] and the size of ventral prostate to around 0.05% (p<0.01). Histological analysis revealed anti-proliferative signs such as reduction in number of acini [(7.50) (p<0.01)], epithelial height [(10.55μm) (p<0.01)] and epithelial acinar area [(18.17%) (p<0.01 )]. Since TUNEL assay showed non-significant outcome which ruled out apoptosis, aqueous methanol extracts have arrested cell cycle by downregulate (p<0.01) proliferative marker,Ki-67 and PCNA. Prostate cell growth is arrested by downregulation of AR which inhibited AR mRNA transcription (RTPCR analysis) and induced cell cycle arrest at S phase through p27 and cyclin E2 (western blot analysis). Paddy husk is a better chemopreventive agent than paddy straw against prostate disease.
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Keywords
Effects Of Paddy , Waste Products , Prostate , Vitro , In Vivo Model