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Study On Anti-tumor Effection Of Lavender Angustifolia Mill. Effective Components And Their Mechanism

Posted on:2013-07-28Degree:MasterType:Thesis
Country:ChinaCandidate:X L SongFull Text:PDF
GTID:2234330374992217Subject:Biochemistry and Molecular Biology
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Objective:To investigate the effects of Lavender angustifolia differrent effective components on HepG-2cell line in vitro, H22carcinoma implanted Kunming mice in vivo and their mechanism.Methods:After HepG-2cell line treated with different dossage of19-hydroxyl WuSu ester, anti-proliferation was assayed by MTT and Flow cytometry assay, LDH activities in culture serum were detected, the morphological changes had observed by HE staining. Agarose gel electrophoresis and Annexin-V and PI coupling staining were carried out to evaluate apoptosis phenomenon and apoptosis rate, respectively. The expression of Bax. BCL-2and anti-proliferation effect of19-hydroxyl WuSu ester on HepG2cell line were analysed by western blot. Tumor inhibition rate, thymus and spleen index of H22carcinoma implanted mouse were observed; TNF-a. IL-6and VEGF in H22carcinoma implanted mouse were assayed by double-sandwich ELISA and leukocyte count was measured. Agarose gel electrophoresis, western blot analysis were carried out in H22carcinoma implanted mouse to observe the anti-proliferation effects and the expression of Bax and Bcl-2.The results:19-hydroxyl WuSu ester plays an obviously role in the inhibition function to HepG-2in vitro. The inhibition rate was46.2%, approximately IC50while treated with6.25μg/mL for24h., HepG-2cell volume decreasing and cell nuclear became heavy staining had observed after different dossage of19-hydroxyl WuSu ester treatment. LDH relative activities were1576.89,1561.61,1884.406. and1411.12U/gprot while using25.00,12,50,6.25and3.13μg/mL of19-hydroxyl WuSu ester treated for24h. respectively. The results of DNA ladder showed obviously apoptosis phenomenon and the apoptosis rate was10.20%,10.85%and32.89%, respectively; the anti-19-hydroxyl WuSu ester serum also showed the inhibition effects to HepG-2cellls. Western blot results indicated that19-hydroxyl WuSu ester up-regulate/down-regulate the expression of Bax/Bcl-2protein. The tumor inhibition rate caused by dosage of Lavender angustifolia ethanol extracts were higher than21%. it obviously improved thymus index, increase the expression levels of TNF-a and IL-6, decrease the levels of VEGF, also:Leukocyte count, there was no significantly difference between Lavender angustifolia ethanol extracts treated group and control group, while there was significantly higher than cyclophosphamide treated group (positive control). The apoptosis also could be observed by DNA ladder, and the Western blot analysis also showed that Lavender angustifolia ethanol extracts could up-/down-regulate Bax/Bcl-2expresion. respectively, in H22carcinoma implanted mouse.Conclusion:The effective components of Lavender angustifolia play an important role in anti-proliferation in HepG-2cell in vitro, and have obvious anti-tumor effects in H22carcinoma implanted mouse in vivo. The mechanism maybe related to regulating immuno-system. inhibiting angiogenesis of tumor tissue, promoting cell necrosis, inducing apoptosis and result in antitumor.
Keywords/Search Tags:Lavender angustifolia, anti-tumor, cytokine
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