الفهرس 
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Abstract
Background & Aim: Hepatocellular carcinoma (HCC) is regarded as one of the most life- threatening cancers, as it is the sixth most common tumor and the third reason for its death worldwide. It is a noteworthy public health concern in Egypt, because of several environmental and host-related risk factors. Chemotherapeutic drugs have disadvantages as toxicity for non-cancerous tissues, drug resistance, and recurrence of cancer. Medicinal plants with their active components have great attention as an important source for novel drug discovery due to their availability, efficiency, and safety. Searching for new strategies to obtain new drugs with higher efficiency and more safety represents an urgent need. One of these strategies is the laser light treatment for seeds, which is known to improve germination, plant growth, and biological activities.
Therefore, the object of the present work was pursued to investigate the effect of laser pretreatment at different powers and time intervals of Balanites aegyptiaca (B. aegyptiaca) seeds on enhancing the germination, phenolic content and antioxidant activity of the methanolic extracts of their whole dry plant material (roots and shoots) through different assays and select the most powerful laser pretreatment extracts. Then the preemptive and therapeutic influences of the most efficient methanolic extracts of roots and shoots of B. aegyptiaca were elucidated on liver tumorigenesis whether in vitro on HepG2 cell lines or in vivo as induced by diethylnitrosamine (DENA)/carbon tetrachloride (CCl4) in mice, in an attempt to prevent, reverse or restrict the cancer development through controlling its different mechanisms.
The experimental design: Firstly, the seeds of B. aegyptiaca were divided into nine groups (50 seeds for each): the control group without any irradiation and eight He-Ne laser treatments conducted at different powers of 25, 50, 100, and 200 mW with two-time intervals for each power at 2 and 4 minutes.
Secondly, there are 8 sets of male CD-1 Swiss albino mice, with 10 animals in each group, were established as follows: Control group was administered vehicle only, HCC group was administered the carcinogen with the promoter of tumorigenesis (DENA/CCl4), 2 extract (LRC and LSC) groups were administered only the roots and shoots extracts whose seeds exposed to 200 mW/4 min (LR and LS) as the most effective laser pretreatment extracts, respectively, 2 prophylactic (LRB and LSB) and therapeutic (LRA and LSA) groups were administered the extracts before and after the DENA/CCl4, respectivly. The experiment was prolonged for 16 weeks.
Results: The results bring to light that the most efficient laser treatment for seeds of B. aegyptiaca was at 200 mW/4 min, which induces the highest yield percentages, total phenolic and flavonoids contents, metal chelating, reducing power, as well as free diphenylpicrylhydrazyl (DPPH) and 2, 2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radicals scavenging activities. Based on these outcomes, the antiproliferative screening assay of the methanolic extracts for the shoot (S) and root (R) dry plant material of B. aegyptiaca after He-Ne laser pretreatment of seeds at 200 Mw for 4 min. (200/4 LS and 200/4 LR) compared with control was performed in vitro on a panel of three cancer cell-lines (HepG-2, HCT-116, and MCF-7) using the sulphorhodamine-B (SRB) assay, and the cytotoxicity was determined using normal BHK fibroblast cell line. Obtained results indicated that these extracts should be regarded as potential anti-carcinogenic resources against the HepG-2 cell line, displayed moderate activity against MCF-7 and HCT-116 cell lines, and exhibited no activity against the growth of the normal BHK cell line.
Based on the aforementioned phytochemical analyses, antioxidant capacity and anticancer activities of the tested plant extracts on different cell lines, HPLC and GC profiles attributed these outstanding properties to various phenolic compounds of LR and LS extracts, which can provide preliminary indication of the potential use of both of them as promising chemo-preventive and/or chemotherapeutic agents in vivo against diethylnitrosamine (DENA)-induced hepatocellular carcinoma (HCC) in mice. The phytochemical compounds whether phenolic acids, flavonoids or others that identified by HPLC and GC-MS profiles discriminate the root extracts from shoot extracts, and this was obviously manifested in our various antioxidant activities, biochemical and molecular analyses.
The biochemical parameters in serum such as liver enzymes (ALT, AST and ALP), kidney function markers (Urea and Creatinine), oxidative stress (TAC, as well as in tissue as MDA and NO), HCC marker (serum AFP), angiogenesis marker (VEGF), tumor growth marker (uPA), autophagy marker (LC3), in addition to molecular biology parameters through real time-PCR for NF-κB p65 as an inflammation marker, Cyt C and Casp-3 as apoptotic markers, and MMP-2 and MMP-9 as metastatic markers were all evaluated and their levels increased significantly in consequence of DENA/ CCl4 in HCC group compared to control group, except TAC and LC3 where their levels decreased significantly in consequence of DENA. Administration of the LR and LS in prophylactic and therapeutic groups caused a significant decrease in such parameters when compared with DENA group, except TAC and LC3 levels where their levels increased significantly in consequence of LR and LS extracts. This indicated that the present extracts have the capacity of suppressing neoplastic alteration by restoring hepatic and nephro-toxicity, relieving the oxidative stress, maintaining AFP status, inhibiting inflammation, halting angiogenesis, stimulating autophagy, reducing apoptosis, prohibiting tumor growth and arresting matrix degradation and subsequently the tumor invasiveness and metastasis in coincide with improvement in the TEM examination, as well as macroscopic and histopathological alterations. Furthermore, the present extracts had more protective action than therapeutic action as their actions on almost all parameters were more effective in prophylactic (LRB and LSB) groups than therapeutic (LRA and LSA) groups. However, roots methanolic extracts were roughly more outperformed than their counterparts of shoots extracts. Conclusion: The current extracts might be a promising candidate for being chemopreventive substances versus both the liver tumorigenesis and the disease progression, but these vital extracts could have more prophylactic action than therapeutic action.