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The Study Of The Effect And Mechanism Of Compound Phyllanthus Urinaria (CPU) And Artemisia Annua Ethanol Extract On Hepatoma

Posted on:2012-10-23Degree:MasterType:Thesis
Country:ChinaCandidate:J PeiFull Text:PDF
GTID:2154330335468032Subject:TCM clinical basis
Abstract/Summary:PDF Full Text Request
1 ObjectivesThe Compound Phyllanthus Urinaria(CPU) was made by screening and optimizing its components for many years, basing on the theory of Chinese medicine. CPU was composed of ye xia zhu(Phyllanthus Urinaria), lv cha (Green Tea), san qi(panax notoginseng), and ye xia zhu was the major component. The compound recipe have been demostrated that have the better effectiveness of soothing liver, strengthening spleen, regulating vital energy, detoxication, promoting blood circulation and removing blood stasis. In recent years, the research of Phyllanthus Urinaria has made great progress at home and abroad. In addition for the treatment of liver dysfunction caused by the hepatitis B, the literature has been reported Phyllanthus Urinaria can inhibit tumor growth. Early research has initially confirmed that CPU has the effect of anti-hepatoma.Qing hao(Artemisia annua) is Compositae annual herb, also known as Artemisia argyi and Artemisia annua L, The traditional effect of qing hao is clearing deficient heat and summer heat,preventing malaria. It is used for the treatment of summer heat, deficient heat, spastic muscles,achy joints with hot flashes, malarial disorders. Artemisia annua was composed of many components,including Volatile oil, sesquiterpenes, flavonoids and coumarin. Artemisinin belongs to sesquiterpene lipid compounds, and it is main active ingredient in Artemisia annua. In addition for the treatment of anti- malaria, now there are many research on anti-tumor effect of Artemisinin and its derivatives. In addition to artemisinin, whether there are other anti-tumor component in the Chinese medicine Artemisia annua? Artemisia annua Ethanol extract was a Chinese medicine preparation extracted from Artemisia annua leaves. This experimental research is to study the anti-hepatoma activity of Artemisia annua Ethanol extract.This experimental research is to study the anti-hepatoma effect of Compound Phyllanthus Urinaria and Artemisia annua Ethanol extract in mice of transplanted hepatocarcinoma22(H22), and study the influence of these two drugs on tumor-bearing mice serum IL-2 and TNF-αlevels; also study the influce of CPU on the proliferation of HepG2 and to explore the mechanism of anti-tumor at cell level. It aimed to offer scientific data for further research on CPU and Artemisia annua Ethanol extract.2 Methods2.1 To study the effect and mechanism of CPU on hepatoma2.1.1 To study the inhibitory effect and mechanism of CPU on tumor growth on murine transplanted hepatocarcinoma 22(H22)in vivoEstablish mice model of hepatocarcinoma 22(H22). The tumor bearing mice were randomly divided into control group, cyclophosphamide group(CTX), high dosage of CPU, middle dosage of CPU, low dosage of CPU,each group contained 8 mice. After 24h by inoculating tumor cells, These mice were given different drugs for 8 days, Tumor growth state was observed and tumor inhibitory rate was calculated 24h after the last administration; enzyme-linked immunosorbent assay(ELISA) was used to detect IL-2 and TNF-a levels in tumor-bearing mice serum.2.1.2 To study the effect of CPU on inhibiting proliferation of HepG2HepG2 was cultured and divided into Control group, cell group and different dosage of CPU, MTT colormetric assay was used to evaluate the inhibitive effect on HepG2.2.2 To study the effect and mechanism of Artemisia annua Ethanol extract on hepatoma2.2.1 Extraction of Artemisia annua Ethanol extract and quantitative determination of ArtemisininTo Extract Artemisia annua Ethanol extract from Artemisia annua leaves with 60% ethanol, and To establish an HPLC method for the quantitative determination of the Artemisini in Artemisia Ethanol Extract.2.2.2 To study the inhibitory effect and mechanism of Artemisia annua Ethanol extract on tumor growth on murine transplanted hepatocarcinoma 22(H22)in vivoEstablish mice model of hepatocarcinoma 22(H22). The tumor bearing mice were randomly divided into control group,5-fluorouracil group (5-FU), low dosage of Artemisia Ethanol Extract, high dosage of Artemisia Ethanol Extract, each group contained 8 mice. After 24h by inoculating tumor cells, These mice were given different drugs for 8 days, Tumor growth state was observed and tumor inhibitory rate was calculated 24h after the last administration; enzyme-linked immunosorbent assay(ELISA) was used to detect IL-2 and TNF-a levels in tumor-bearing mice serum.3 Results and conclusion3.1 CPU can significantly inhibit hepatocarcinoma growth, Also it can increase IL-2 level and decreased TNF-a level in tumor-bearing mice serum. Its inhibitory mechanism is concerned with regulating the body's immune response by increasing IL-2 level and decreasing TNF-a level.In vitro experiment, it showed that Compound Phyllanthus Urinaria can significantly inhibit the proliferation of HepG2 cells. Inhibition rate correlated with drug concentration, showing a clear dose-effect and time -effect relationship. These results showed that the inhibitory mechanism of Compound Phyllanthus Urinaria is concerned with inhibition of tumor cell proliferation.3.2 Artemisia Ethanol Extract can significantly inhibit hepatocarcinoma growth. The Preliminary experimental results showed that Artemisia Ethanol Extract decreased TNF-a level in tumor-bearing mice serum. Its inhibitory mechanism is concerned with regulating the body's immune response by decreasing TNF-a level.In addition, this study also determined the quantitation of the Artemisini in Artemisia Ethanol Extract, the determination was 0.02%. However, in the experiment of effect of Artemisini on tumor growth in mice of transplanted hepatocarcinoma 22(H22), Artemisini(10mg·kg-1·d-1) can not inhibit hepatocarcinoma growth. In this study, Artemisini is not major component of anti-tumor, In addition to Artemisinin, other anti-tumor components may exist in Artemisia annua. The specific anti-tumor component is worthy to be further studyed.
Keywords/Search Tags:Compound Phyllanthus Urinaria, Artemisia Ethanol Extract, hepatocarcinoma, tumor inhibitory mechanism, tumor inhibitory rate
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