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Pomegranate root bark and other 9 kinds of traditional Chinese medicine in vitro anti-human observation of the effect of Blastocystis hominis
Posted:2008-5-25 15:11:00 Browse:14745 Times
Author: Su Shui-lian, Chen, Guifeng, Zhang Rui their, XIE Qiong
Key words Blastocystis hominis;,, pomegranate root bark;,, Picrorhiza;,,,, insecticidal effects;,,,, form
Abstract: Objective To investigate the root bark of pomegranate and other 9 kinds of Chinese herbal medicine to kill the effect of Blastocystis hominis. Methods of different concentrations of water extract of Chinese herbal medicine in vitro anti-human Blastocystis hominis experiment, and light microscope observation of the role of future generations morphology of Blastocystis hominis. Results With the extension of time drug action and drug concentration increased, the root bark of pomegranate, Hu Huang Lian, Coptis chinensis, Meliae pairs of Blastocystis hominis with different degrees of apoptosis, these drugs against Blastocystis hominis in MIC values were 0.1,0.6,0.8,0.8 g / ml, light microscopic observation
parasite surface membrane depression, exfoliation, rupture. Low concentration of
parasites after drug effect picrorhizae are transformed into cysts, while the Chinese bulbul, pumpkin seeds, so that a gentleman, betel nuts, matrine for the role of Blastocystis hominis no. Conclusion pomegranate root bark, Picrorhiza directly act on the
parasite surface membrane, with a strong killing effect of Blastocystis hominis.
Key words: Blastocystis hominis; pomegranate root bark; Picrorhiza; insecticidal effect; morphology
Abstract: ObjectiveTo study the effect of Psidium guajaual and other eight Chinese herbs on the Blastocysitis hominis. MethodsTrchomonacidal effect of Chinese herbs was investigated using various concentration of Chinese herbs water extract against Blastocysitis hominis in vitro.The morphological changes of the Blastocysitis hominis were observed by light microscope.ResultsWith prolongation of the time of drug action and increase of its concentration, Psidium guajaul, Picrorhra scrophulariitlora Pennell, Coptis chiennsis Franch and Melia azedarach Linn have the effect of apoptosis to Blastocystis hominis. Their lowest concentrations were respectively 0.1,0.4,0.6 , 0.8g/ml.We can observe the cell membrance hollowed, exfoliated and ruptured. The Blastocystis hominis change into cyst with low concentration of Picrohica scrophulariitlora Pennell. And Pulsatilla chinenensis Bunge Regel, Cucurita moschata Ouch, Quisgualis indical, Areca actechata Duch and Sophora flavescens Aic have no effect to Blastocystis hominis.ConclusionPsidium guajaual and Picrorhica scrophulariitlora Pennell destroyed cell membrane directly and exhibited effective trichomonacidal Blastocystis hominis action.
Key words: Blastocystis hominis; Psidium guajaual; Picrorhica scrophulariitlora Pennell; Trichomonacidal effect; Objective conformation
Blastocystis hominis (Blastocystis hominis short Bh) in the last 20 years has been valued by people with diarrhea-related human parasitic protozoan. In our country, which have been classified as emerging infectious diseases caused by the disease [1], Blastocystis hominis is generally represented by scattered onset, but in southern Jiangxi Chongyi County occurred in 1996, an outbreak of epidemic [2]. 2002 in our hospital infection rate was 10.09% newborn Bh (426/43) [3]. We have used metronidazole treatment for those infected and 3 months after the review of most of the overcast results are poor, and easy to recur. Study further showed that metronidazole has a teratogenic and carcinogenic effects [4]. Reported in the literature of the root bark of pomegranate sour, astringent taste, with astringent intestinal bleeding, the efficacy of insecticide [5]. The main treatment bacillary dysentery, amoebic dysentery and a variety of infectious diseases. Picrorhiza insecticidal efficacy of such a detoxification [6]. In order to provide the development of new antiprotozoal herbs experimental data, we used nine kinds of Chinese herbal medicine in vitro killing effect of Bh. Are reported below.
1 Materials
1.1 Bh strains of insect pest strains isolated from Affiliated Hospital of Gannan Medical College of Gastroenterology, pediatric patient's faeces (36 �� 0.5) �� general anaerobic incubator, every 3 days passaged one time, a total of 20 times passage medium.
1.2 Drug experimental drugs (purchased from affiliated hospitals, hospital pharmacies and the local pharmaceutical companies to retail outlet), all for Chinese medicine, the book has the effect of heat-clearing and detoxifying and de-worming of Chinese herbal medicine.
1.3 medium Locke's egg serum (LockeEggSerum, LES) two-phase medium [5], into a 30 �� bevel set 90 ~ 100 �� oven made of heat coagulation. Before switching to each tube add 5 ml Rockwell liquid, sterile 14% inactivated newborn calf serum (purchased from Hangzhou, China, Chinese holly leaf Biological Engineering Materials Co., Ltd.) and the basic
Antibiotic solution (0.4% ampicillin, 0.1% chain mycin, 0.000 6% amphotericin B) can be transferred kinds.
2 Methods
2.1 Preparation of liquid would be nine kinds of Chinese herbal medicines (dried drug) of the 10 g, respectively, into 250 ml triangular flask in each of the home 50 ml distilled water, soak overnight, then put into 80 �� water bath in the hot-dip 4 h. Filtered to remove dregs, the filtrate were concentrated by gentle heat to the liquid concentration of 1,4,6,8. 3 000 r / min centrifugation 10 min, the supernatant obtained, leaching sterilization, adjust the osmotic pressure, set sterile bottle , 4 �� to save backup.
2.2 The optical microscope observation of each type of herbal drug concentration in the preparation of four groups, respectively, 0.1,0.4,0.6,0.8 g / ml, at the same time set up drug-free control group. Remove the culture of Blastocystis hominis 36h culture tubes, respectively, from cultured basal layer suction equal 20 ��l
parasite suspension drops on the glass slide, add coverslip microscope. To do with the blood cell counter counting observations, to calculate the number per mm3 contained in the worm. The concentration of
parasites in each tube is about 1 �� 103 Ge / ml, and then 1ml by adding different concentrations of liquid in the culture tubes placed (36 �� 0.5) �� incubator. Cultured 2,4,8,12 h respectively after drawing the tube worm drops film microscopy, using high-power microscope parasite active, calculate
parasites mortality. Cultured 24 h after the mortality rate was 100% of the experimental tube switching to, (36 �� 0.5) �� further incubated 48 h, observe whether there is Bh survival, all the experiment was repeated 3 times. Reposted elsewhere in the paper for free download
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3 Results
The same role of the time, the root bark of pomegranate, Hu berberine, berberine, Cortex Meliae group Bh mortality rate were higher (P <0.01), and the parasite mortality rate increased with increasing drug concentration (P <0.01); the same drug concentration, with the role of prolonged, Bh mortality gradually increased (P <0.05). High-powered microscope, the role of pomegranate root bark of group 2 h after, 1,4,6,8 g / ml concentration group, whole parasite apoptosis, morphogenesis significant changes. Most of the parasite membrane edges blurred, parasites
Depression cracking (Figure 1). The role of picrorhizae group 2 h after 1,4 g / ml concentration group, whole parasite-induced into a cyst (Figure 2). The role of 8 h, 6,8 g / ml concentration group, killing all parasites. Then pomegranate root bark, and Picrorhiza kinds of cultured insect medium group transferred 24 h, were no Bh growth. Chinaberry bark, Coptis the role of the parasite is not strong, while the Chinese bulbul, pumpkin seeds, so that a gentleman, betel nuts, matrine on Blastocystis hominis no effect. The results in Table 1. Intact parasite control group (Figure 3).
Figure 1 (omitted)
Figure 2 (omitted)
Figure 3 the control group (normal cultured human Blastocystis hominis) (omitted)
MIC determination: The results showed that the root bark of pomegranate, Picrorhiza apoptosis of human Blastocystis hominis fastest, shortest, MIC minimum. The Coptis, Chinaberry skin effect is not strong, betel nuts, Sophora flavescens, Pulsatilla, pumpkin seeds, so that a gentleman on the role of Blastocystis hominis no. The results in Table 2.
Table 19 kinds of Chinese herbs to kill Blastocystis hominis in vitro results (omitted)
Table 29 kinds of Chinese herbal B.h experiments MIC comparison (omitted)
4 Discussion
Pomegranate root bark of Psidium guajaual traditional Chinese medicine is a deciduous shrub or small tree, with astringent antidiarrheal intestinal bleeding, the efficacy of insecticide detoxification. Commonly used in the treatment of clinical Jiu Li, Jiuxie, hematochezia, prolapse, uterine bleeding, vaginal discharge, abdominal pain, insect plot embolism [6]. In recent years, studies have shown that pomegranate roots leather are right there are a variety of bacteria and influenza virus inhibition [7]. Picrorhiza is a multi-year growth of low vegetation Scrophulariaceae plants, is a commonly used medicinal herbs. There is heat, dehumidifiers, eyesight, Bugan the ECM gallbladder, insecticidal effectiveness. This study confirmed that the pomegranate has obvious roots leather to kill the role of B. hominis, 0.1 g / ml concentrations of drug action 2 h of the Bh all have to kill and rifting. Picrorhiza encysted parasites can be induced early, as drug concentration increases, the role of the extension of the time, Bh mortality rates have increased, that is insecticidal effect and drug concentration and effect of time into a positive correlation. 0.6 g / ml concentrations of drug action 8 h all of apoptosis in B. hominis. And, after switching to no B.h growth. Select the appropriate drug concentration and the role of time for light microscopic observation to study the mechanism of drug action key. Accordingly, this experiment use a low concentration of the root bark of pomegranate liquid, parasites rapid apoptosis, pyrolysis, it shows that the insecticidal effect of the root bark of pomegranate, but can not observe the parasite surface membrane damage to the dynamic process of change, the exact mechanism deserves further study. Low concentration of Picrorhiza liquid drug can be fully observed by the changing role of parasites, trophozoites induced into the first sac, and then in high concentrations of drug effects, the parasites die rapidly. Coptis, Chinaberry Peel on the role of Blastocystis hominis is not strong, while the Chinese bulbul, pumpkin seeds, so that a gentleman, betel nuts, matrine on Blastocystis hominis had no effect, it is that the root bark of pomegranate, water extract of Picrorhiza have to kill The role of Blastocystis hominis. Pomegranate root bark, Picrorhiza is a common Chinese herbal medicine, China's resource-rich, low cost and low toxicity. This experiment for the use of pomegranate root bark, Picrorhiza the development of new high-efficiency low-toxic topical and oral anti-drug Blastocystis hominis basis.
References:
[1] Pan-chang. Emerging Infectious Diseases [M]. Beijing: People's Health Publishing House, 2004,153.
[2] Wu Wang, bear in tree, Cao Gao Lu, et al. Blastocystis hominis with the outbreak investigation and research [J]. Chinese Journal of Parasitology Journal of Prevention 2000,13 (1): 25.
[3] Su Shui-lian, Chen, Guifeng, Teng Min-chang, et al. College freshmen survey of intestinal parasitic infection [J]. Chinese Journal of Parasitology Prevention magazine, 2003,16 (6): 381.
[4] Goldman P. Metronidazole: proven benefit and potential risks [J]. Jones Hopkins MED J, 1980,147:1.
[5] Su Shui-lian, Yan Yi-ming, Chen, Guifeng, et al. Three kinds of culture media in vitro effect of Blastocystis hominis observations [J]. Chinese Journal of Parasitology Journal of Control, 2004,17 (6): 375.
[6] Yang Liping, Yang Yonghong. Pomegranate husk of the research [J]. Yunnan Journal of Traditional Chinese Medicine, 2004,25 (3): 45.
[7] Jiangsu New Medical College. Dictionary of Traditional Chinese Medicine [M]. Shanghai: Shanghai
Science and Technology Publishing House, 1996:1262. Reposted elsewhere in the paper for free download
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