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Pomegranate root bark and other 9 kinds of traditional Chinese medicine against B. hominis in vitro effect of observation
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On: Su Shui-lian, Chen Guifeng, Zhang Rui its Xie Qiong

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Key words hominis;,, pomegranate root bark;,, Picrorhiza;,,, insecticidal effect;,,, 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. Method using different concentrations of herbal extracts in vitro experiments against B. hominis, and the role of light microscope descendants hominis morphological changes. The results with the drugs time and the concentration of drug increased, the root bark of pomegranate, Picrorhiza, berberine, Melia hominis skin on different levels of apoptosis, these drugs on B. hominis in the MIC values were 0.1,0.6,0.8,0.8 g / ml, light microscopy Depression parasite surface membrane, stripping, broken. Low concentration picrorhizae both drugs after the larvae change into cysts, and Weng, pumpkin seeds, make a gentleman, betel nut, matrine on hominis had no effect. Conclusion pomegranate root bark, Picrorhiza can act directly on parasites surface membrane, and a strong role in killing B. hominis.

Key words: Blastocystis hominis; pomegranate root bark; Picrorhiza; insecticidal effect; form

The Trichomnacidal Effect of Nine Chinese Herbs on the Blastocystis hominis

SU Shui lian, CHEN Gui feng *, ZHANG Rui qi, XIE Qiong jun, LIAO Hua

(Gannan Medical College, Ganzhou, Jiangxi Province 341000, China)

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

B. hominis (Blastocystis hominis short Bh) in the last 20 years has been valued by people with diarrhea-related human parasitic protozoa. In our country, has as its emerging infectious diseases caused by the disease [1], people generally Blastocystis protozoal disease was sporadic, but in Southern Chongyi County occurred in 1996 outbreak [2]. 2002 students in our hospital infection rate was 10.09% Bh (426/43) [3]. We have those infected with metronidazole treatment, 3 months later most of the negative conversion results are poor, and easy to relapse. Another study shows that metronidazole is teratogenic and carcinogenic effects [4]. Pomegranate root bark of the reported acid, astringent taste, with astringent intestinal bleeding, the effect of pesticides [5]. The main treatment of bacillary dysentery, amoebic dysentery and a variety of infectious diseases. Picrorhizae insecticidal efficacy of such a detoxification [6]. To provide, develop new herbal insect antigens experimental data, we used nine kinds of Chinese herbal medicine in vitro killing effect of Bh. Are reported as follows.

1 Materials

1.1 Bh strains strains isolated from Department of Gastroenterology, Affiliated Hospital of Gannan Medical College, pediatric patients with faeces (36 ± 0.5) ℃ common anaerobic incubator, passaged every 3 days 1, a total of 20 times passaged medium.

1.2 Drug test drug (purchased from the hospital pharmacy in hospitals and the local subsidiary of pharmaceutical company retail outlet), all for the book, clearing and detoxifying Chinese medicine and insecticide effects of Chinese herbal medicine.

1.3 Locke's egg medium serum (Locke Egg Serum, LES) two-phase medium [5], into a 30 ° bevel 90 ~ 100 ℃ oven for home heating solidification made. Before switching to each tube add 5 ml Rockwell liquid, sterile 14% inactivated newborn calf serum (purchased from Hangzhou, China, holly leaf Biological Engineering Co., Ltd.) and basic Antibiotic solution (0.4% ampicillin, 0.1% chain ADM, 0.000 6% amphotericin B) can switch species.

2 Methods

2.1 Preparation of the nine kinds of Chinese herbal liquid (dry drug) of the 10 g, were loaded into 250 ml triangular flask, 50 ml distilled water in each home, soaked overnight, then put into 80 ℃ water bath in the hot dip 4 h. Filter to remove residue, filtrate were concentrated with a slow fire to the liquid concentration of 1,4,6,8. 3 000 r / min centrifugation 10 min, the supernatant obtained, filtration sterilization, adjust the osmotic pressure, and placing sterile bottle , 4 ℃ save backup.

2.2 The optical microscope for each preparation of four herbs drug concentration group, respectively, 0.1,0.4,0.6,0.8 g / ml, while no drug control group design. Remove the 36h culture of B. hominis culture tubes, respectively, from cultured basal layer suction equal 20 μl drops of parasite suspension in a slide, add coverslip microscope. Blood cell count observation register to do to calculate the number per mm3 contained in the worm. Each tube worms concentration of about 1 × 103 个 / ml, then adding different concentrations of liquid 1ml each culture tube, place (36 ± 0.5) ℃ in an incubator. 2,4,8,12 h culture were drawn after the droplet slides examined each tube worms, worms with active high power microscope to calculate worm mortality. After 24 h, mortality was 100% of the test tube transfer species, (36 ± 0.5) ℃ cultured for 48 h, see if there Bh survival, all the experiment was repeated 3 times.

3 Results

The same action time, pomegranate root bark, Picrorhiza, berberine, Melia Paper Group Bh mortality rate higher than the control (P <0.01), and larvae mortality increased with increasing drug concentration (P <0.01); the same drugs concentration, with the action time, Bh mortality gradually increased (P <0.05). High magnification, the role of pomegranate root bark group after 2 h, 1,4,6,8 g / ml concentration group all worms apoptosis, morphological change significantly. Most worms membrane edge blur, body sag cracking (Figure 1). The role of picrorhizae group 2 h after the 1,4 g / ml concentration group all induce parasite cysts (Figure 2). 8 h, the role of 6,8 g / ml concentration group worms all died. Then the root bark of pomegranate and Picrorhiza species of insect culture medium group transferred 24 h, showed no Bh growth. Neem bark, Coptis effect on the insect body is not strong, and Weng, pumpkin seeds, make a gentleman, betel nut, matrine on B. hominis had no effect. The results in Table 1. Worms intact control group (Figure 3).

Figure 1 (omitted)

Figure 2 (omitted)

Figure 3 the control group (normal cultured B. hominis) (abbreviated)

MIC determination: The results showed that the root bark of pomegranate, Picrorhizae apoptosis hominis fastest, shortest, MIC minimum. And Coptis, Melia skin effect is not strong, betel nut, flavescens, Pulsatilla, pumpkin seeds, make a gentleman of the B. hominis had no effect. The results in Table 2.

Table 19 kinds of Chinese herbs Blastocystis hominis in vitro killing effect (abbreviated)

Table 29 kinds of Chinese herbs Bh MIC experiment comparison (omitted)

4 Discussion

Pomegranate root bark of Psidium guajaual medicine is deciduous shrub or small tree, intestinal diarrhea with astringent to stop bleeding, the efficacy of insecticide detoxification. Commonly used in clinical treatment of Jiu Li, Chronic Diarrhea, hematochezia, rectal prolapse, bleeding, vaginal discharge, abdominal pain embolism product insects [6]. Recent studies show that there is a wide range of leather pomegranate root bacteria and inhibition of influenza virus [7]. Scrophulariaceae Picrorhiza is the growth of low vegetation and plants for many years, is a commonly used medicinal herbs. With heat, dampness, eyesight, Bugan, Yidan, insecticidal effectiveness. This study confirmed that pomegranate root kill leather obvious role of B. hominis, 0.1 g / ml concentration of the drugs were 2 h with the killing of the Bh and decomposition. Picrorhizae worms into the capsule can be induced early, with the increase in drug concentration, treatment time, Bh increased mortality, that is insecticidal effects and drug concentrations and times of a positive correlation. 0.6 g / ml concentrations of drugs hominis 8 h all of apoptosis. And after switching to no Bh growth. Select the appropriate drug concentration and time were observed by light microscopy is a key mechanism of drug action. Accordingly, this experiment used a low concentration of the root bark of pomegranate liquid, body fast apoptosis, cleavage, demonstrated the killing effect of the root bark of pomegranate, but can not observe the parasite surface membrane damage dynamic process, its exact mechanism worthy of further study. Low concentration of liquid to fully observe picrorhizae worms by the drug effects of changes in trophozoites induce capsule first and then the high concentration of drugs after the larvae quickly die. Coptis, Melia Paper on the role of B. hominis is not strong, and Weng, pumpkin seeds, make a gentleman, betel nut, matrine on B. hominis had no effect, so that the root bark of pomegranate, water extract of Picrorhiza have to kill buds hominis role. Pomegranate root bark, Picrorhiza is a common Chinese herbal medicine, China's resource-rich, low cost and low toxicity. The experiment for the use of pomegranate root bark, Picrorhiza development of new high-performance topical and oral anti-toxicity drugs hominis basis.

References:

[1] Pan Xiaozhang. Emerging Infectious Diseases [M]. Beijing: People's Medical Publishing House, 2004,153.

[2] Wu Guohong, bear in tree, Cao Gao Lu, et al. With human Blastocystis protozoal outbreak investigation and study [J]. Chinese Journal of Parasitology Control 2000,13 (1): 25.

[3] Su Shui-lian, Chen Guifeng, Teng Min Chang, et al. Freshmen Survey of intestinal parasitic infection [J]. Chinese Journal of Parasitology Control, 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 Yiming, Chen Guifeng, et al. Three kinds of culture medium in vitro effects of B. hominis observation [J]. Chinese Journal of Parasitology Control, 2004,17 (6): 375.

[6] Yang Liping, Yang Yonghong. Granatum of research [J]. Journal of Traditional Chinese Medicine, 2004,25 (3): 45.

[7] Jiangsu New Medical College. Chinese Dictionary [M]. Shanghai: Shanghai Science and Technology Press, 1996:1262.

(Jiangxi Province, Gannan Medical College, Jiangxi 341000)