Effects of Ethyl Acetate Extract of Phyllanthus reticulatus Leaves on Autophagy-related Proteins Beclin-1,ATG5 and LC3 in Nude Mice Xenograft Model of HCC

[Objectives]To explore the effects of ethyl acetate extract of Phyllanthus reticulatus leaves on autophagy-related proteins Beclin-1,ATG5 and LC3 by immunohistochemistry,and to preliminarily explore their effects on autophagy.[Methods]BEL-7404 Hepatocellular Carcinoma(HCC)nude mice model was established,and blank group(same volume of pure water),positive control group(20 mg/kg fluorouracil),high dose drug group(600 mg/kg),and medium dose drug group(300 mg/kg),and low dose drug group(150 mg/kg)were set up.After 2 weeks of intragastric administration,the nude mice were sacrificed,and the tumor tissues were taken out,processed by immunohistochemistry,and then made into paraffin sections.Photos were taken under an optical microscope(10×40),and evaluation and analysis were performed with the aid of the Image-Pro Plus 6.0 image analysis software.Differences were calculated using SPSS 20.0 software.The effects of drugs on autophagy-related proteins LC3,Beclin-1 and ATG5 were observed.[Results]Compared with the blank group,the medium and high dose groups of ethyl acetate extract of P.reticulatus leaves had the effect of promoting the increase of autophagy-related proteins LC3,Beclin-1 and ATG5(P<0.05).However,there was no significant difference between the low dose group of ethyl acetate extract of P.reticulatus leaves and the blank group(P>0.05).[Conclusions]The ethyl acetate extract of P.reticulatus leaves has a promoting effect on autophagy-related proteins LC3,Beclin-1,and ATG5.

Analyzing Nootropic Effect of Phyllanthus reticulatus Poir. on Cognitive Functions, Brain Antioxidant Enzymes and Acetylcholinesterase Activity against Aluminium-Induced Alzheimer’s Model in Rats: Applicable for Controlling the Risk Factors of Alzheimer’s Disease

Oxidative stress is intensely linked with neurodegenerative disorders, especially Alzheimer’s disease (AD). Searching for medicinal plant with the nootropic activity for controling the development and progression of AD has received extensive consideration. The plant Phyllanthus reticulatus (PR) Poir. is known in Bengali as Panjuli belongs to family Euphorbiaceae. Previous studies have shown the antioxidant, analgesic, anti-inflammatory, etc. activities of this plant. Therefore, the objective of this study was to examine the nootropic effect of ethanolic extracts of Phyllanthus reticulatus (EEPR) on cognitive functions, brain antioxidatant enzymes and acetylcholinesterase activity in aluminium-induced rats of cognitive impairment and oxidative stress. The effects of EEPR fruit (i.e., 100 and 200 mg/kg b.w.) were examined for 30 days and its nootropic effect was determined in aluminium treated Swiss albino male rats by behavioral studies such as Passive Avoidance (PA) test, Rewarded Alternation (RA) test and biochemical studies such as superoxide dismutase (SOD), catalase (CAT), contents of thiobarbituric acid reactive substances (TBARS) and acetylcholinesterase (AChE) activity in rats brain tissue homogenates. In PA test, administration of EEPR fruit (i.e., 100 and 200 mg/kg, b.w.) significantly (P < 0.05, P < 0.01) increased step-through latency (STL) in rats on 30th day with respect to disease control group. The percentage of memory retention (MR) for this test was pointedly (P < 0.05) increased in rats treated with EEPR fruit (i.e., 200 mg/kg b.w.) as compared with disease control group. For RA test, EEPR fruit (i.e., 200 mg/kg b.w.) markedly (P < 0.01) increased the correct responses (CR) in rats on 30th day related to disease control group. In case of this test the percentage of MR was significantly (P < 0.05, P < 0.01) increased in rats treated with EEPR fruit (i.e., 100 and 200 mg/kg b.w.) with respect to disease control group. Administration of EEPR fruit (i.e., 100 and 200 mg/kg b.w.) considerably (P < 0.05, P < 0.01) increased the level of SOD, CAT and expressively (P < 0.05) decreased TBARS level compared to disease control group. Treatment with EEPR fruit (i.e., 100 and 200 mg/kg b.w.) markedly (P < 0.05, P < 0.01) decreased the level of AChE activity to that of disease control group. The present study shows that EEPR fruit has excellent nootropic effect on cognitive performance and brain antioxidant markers in aluminium-induced rats of cognitive impairment and oxidative stress which could be developed in the management of neurodegenerative diseases especially AD.