Asian Pacific Journal of Tropical Medicine

211-220

Authors: Moussa Labsi, Lila Khelifi, Dalila Mezioug, Imene Soufli, Chafia Touil-Boukoffa

Objective To investigate the effect of pomegranate peel aqueous extract (PGE) on the development of secondary experimental echinococcosis and on the viability of Echinococcus granulosus protoscoleces, and the immunomodulatory properties of PGE. Methods Swiss mice were inoculated intraperitoneally with viable protoscoleces. Then, PGE was orally administered daily during cystic echinococcosis development. Cyst development and hepatic damage were macroscopically and histologically analyzed. The production of nitric oxide and TNF-α was assessed in plasma and the hepatic expression of iNOS, TNF-α, NF-κB and CD68 was examined. Moreover, protoscoleces were cultured and treated with different concentrations of PGE. Results It was observed that in vitro treatment of protoscoleces caused a significant decrease in viability in a PGE-dose-dependent manner. In vivo, after treatment of cystic echinococcosis infected mice with PGE, a significant decrease in nitric oxide levels (P < 0.000 1) and TNF-α levels (P < 0.001) was observed. This decline was strongly related to the inhibition of cyst development (rate of hydatid cyst growth inhibition = 63.08%) and a decrease in CD68 expression in both the pericystic layer of hepatic hydatid cysts and liver tissue (P < 0.000 1). A significant diminution of iNOS, TNF-α and NF-κB expression was also observed in liver tissue of treated mice (P < 0.000 1). Conclusions Our results indicate an antihydatic scolicidal effect and immunomodulatory properties of PGE, suggesting its potential therapeutic role against Echinococcus granulosus infection.

228-234

Authors: Ibrahim M. El-Ashmawy, Naser A. Al-Wabel, Aida E. Bayad

Objective To evaluate activity of methanol extract of Achillea fragrantissima (meth) (A. fragrantissima) alone or in combination with diminazine aceturate (DA) against Trypanosoma evansi (T. evansi) in experimentally infected rats. Methods Sixty adult male Wister albino rats were divided equally into 6 groups (A–F). Rats in groups A–E were experimentally infected with T. evansi and those in group F were uninfected. The groups were treated respectively as follows: group A-with 3.5 mg/kg DA; group B- with 1 000 mg/kg meth A. fragrantissima; group C-3.5 mg/kg DA plus 500 mg/kg meth A. fragrantissima; group D-3.5 mg/kg DA plus 1 000 mg/kg meth A. fragrantissima. Group E was left untreated. Parasitemia, survivability, packed cell volume, hemoglobin concentration, total leucocytes count, lymphocyte count, and serum malondialdehyde and reduced glutathione (GSH) levels were estimated. Phytochemical screening of meth A. fragrantissima was also performed. Results The phytochemical analysis of the meth A. fragrantissima indicated a higher content from polyphenolic tannins and non tannins and flavonoids. The efficacy percentage against trypanosomiasis in groups A – E was respectively as follows 80, 40, 90, 100, 0. The administration of meth-A. fragrantissima (1 000 mg/kg b.wt.) produced a moderate efficacy against trypanosomiasis. Untreated rats in group E died between 25 and 30 d post infection. The rats given DA and meth A. fragrantissima combinations (C and D) showed faster and higher recovery rates than the uninfected control and groups A and B. The initial reduction in packed cell volume, hemoglobin, total leucocytes count, increases in serum malondialdehyde and decreases in GSH levels were reversed by the treatments. Conclusion The administration of the methanol extracts of A. fragrantissima and DA combination therapy was more effective than each product alone in the treatment of rats infected with T. evansi and further studies are required to isolate more active ingredients.

241-246

Authors: Rashmee Topno, Siraj Ahmed Khan, Pritom Chowdhury, Jagadish Mahanta

Objective To explore the antiviral activity of antibiotic compounds, mainly aminoglycosides and tetracyclines against Japanese encephalitis virus (JEV) induced infection in vitro. Methods Antiviral activity were evaluated against JEV using cytopathic effect inhibition assay, virus yield reduction assay, caspase 3 level, extracellular viral detection by antigen capture ELISA and viral RNA levels. Results JEV induced cytopathic effect along with reduction of viral progeny plaque formation indicated antiviral potential of the compounds suggesting that antibiotics had broad spectrum activity. Doxycycline and kanamycin administration in dose dependent manner declined viral RNA replication. Conclusions The present study shows kanamycin and doxycycline can affect virion structure and alter replication causing inhibition of JEV induced pathogenesis in vitro.

252-255

Authors: Jeyanthi Suppiah, T.S. Saraswathy, K. Amry, Apandi Yusof, Zainah Saat

Objective To identify the circulating serotypes of human echovirus in Malaysia from 2002 to 2013. Methods A total of 31 retrospective samples from non-polio acute flacid paralysis, hand-food-and-mouth disease, viral meningitis and enterovirus cases were subjected to amplification of partial VP1 gene by RT-PCR. Results Sequencing and phylogenetic analysis of the partial sequences identified presence of human echovirus and human coxsackie viruses. It was found that echovirus 11 was the commonly circulating serotype followed by echovirus 6, echovirus 7, echovirus 3, echovirus 9, echovirus 30 and echovirus 1 in decreasing order. Additionally two types of human coxsackie virus isolates were detected which were coxsackie A24 and B3. Conclusions From the findings, there is a possibility that echovirus 11 is the predominant serotype among Malaysian patients with echovirus infection. However, a larger sample size will yield a more confident result to support this evidence.

261-264

Authors: Wen-Xian Zhang, Jian-Hong Zhao, Fu-Min Ping, Zhi-Jun Liu, Jun-Xia Gu, Xin-Qing Lu

Objective To discuss the effect of dimethyl fumarate (DMF) on rats with l-arginine induced chronic pancreatitis (CP). Methods Male Wistar rats were given DMF treatment (25 mg/kg) by oral lavage method; then Wistar rats were given the intraperitoneal injection of l-arginine for 5 times (250 mg/100 kg, twice per time, each interval of 1 h) for building of CP model. Rats were divided into control group, CP group, DMF group and CP + DMF group. Rats in CP + DMF group were given the oral intragastric administration of DMF (25 mg/kg), while rats in control group and CP group were given the equal volume of normal saline. The weight of rats was evaluated and the intraperitoneal glucose tolerance test was performed (IPGTT, 2 g/kg). The islet of rats was isolated and then flow cytometry was employed to evaluate the quality and activity of islets. Meanwhile, the histology of non-endocrine tissues and levels of myeloperoxidase (MPO) and malondialdehyde (MDA) were detected. Results Compared with control group, the weight of rats in CP group was significantly reduced at week 2, 4 and 6; the blood glucose significantly increased, AUC increased, the histopathological scores of pancreatic atrophy, acinar injury, edema and cellular infiltration increased, levels of MDA and MPO increased, the islet equivalent and islet activity decreased at 0, 30, 60, 120 and 180 min. Compared with CP group, the weight of rats in CP + DMF group significantly increased at week 2, 4 and 6; the blood glucose significantly decreased, AUC decreased, the histopathological scores of pancreatic atrophy, acinar injury, edema and cellular infiltration decreased, levels of MDA and MPO decreased, the islet equivalent and islet activity increased at 0, 30, 60, 120 and 180 min. Conclusions DMF treatment can improve CP induced by l-arginine and islet function in rats.

269-273

Authors: Shi-Xun Wang, Lei Tan, Jian Wang, Jing-Quan Zhong

Objective To study the effect of levocarnitine (L-CN) on tissue inhibitor of metalloproteinase-1 (TIMP-1) and intercellular adhesion molecule-1 (ICAM-1) expression of rats with coronary heart disease and evaluate the protective effect of L-CN on myocardial cells. Methods High-fat diet feeding and intraperitoneal injection of pituitrin was performed on rats in model group and CHD Model of rats was built. Rats with successful model-building were selected and divided into L-CN group and Ctrl group randomly. Rats in L-CN group were given L-CN treatment, with intraperitoneal injection of 200 mg·kg−1·d−1 and successive administration for 3 d. Rats in Ctrl group were given equal volumes of normal saline. Blood was collected from carotid artery at different time and expression quantity of creatine kinase-MB (CK-MB) and Troponin Ⅰ (TnⅠ) in serum was detected. Rats in each group were put to death and were separated to obtain the myocardial tissue. Real-time PCR and Western Blotting hybridization were performed to detect the TIMP-1, ICAM-1 expression in myocardial tissue in each group. Statistical analysis was employed to explore the expression changes of TIMP-1 and ICAM-1, and ELISA test was used to analyze the expression changes of myocardial necrosis marker-CK-MB and TnⅠto learn the effect of L-CN and its myocardial protective effect. Results The total cholesterol, triglyceride and blood glucose levels of rats in model group were significantly higher than that in control group, which indicated that due to high-fat diet feeding, blood lipid of rats in model group was obviously higher than that in control group. In myocardial tissue of rats in model group, TIMP-1 level significantly reduced and ICAM-1 level significantly increased (P < 0.01). In model group, after L-CN treatment, TIMP-1 level had double increase, while ICAM-1 level had 43% of decrease in L-CN group compared with Ctrl group. After L-CN intervention treatment, CK-MB and TnⅠ content in L-CN group relatively reduced compared with Ctrl group. The difference among groups was obvious (P < 0.01). Conclusions L-CN could increase the TIMP-1 expression level and inhibit the ICAM-1 expression level. L-CN has a certain myocardial protective effect.

278-282

Authors: Yan Jiang, Lin Zhang

Objective To explore the mechanism of all-transretinoic acid (ATRA) increasing retinoblastoma (RB) sensitivity to vincristine, and the inhibiting effect of vincristine combined with ATRA treatment on the SO-RB50 cell proliferation. Methods SO-RB50 cells were cultivated by routine culture method. Different concentrations of vincristine or ATRA were added into culture solution. After 48 h, cell counting kit-8 was used to detect the median inhibitory concentration (IC50) of vincristine combined with ATRT treatment to SO-RB50 cells. SO-RB50 cells were divided into drug combination group, vincristine group, ATRA group and control group. Different drugs were added into the culture solution respectively for cell culture based on the IC50 value. Cell counting kit-8 was used to detect the cell proliferation every 24-h cultivation. After continuous determination for 6 d, data was processed to draw the cell growth curve. After drug use for 72 h, flow cytometry was used to detect the proportion of different cell cycles of SO-RB50 cells in each group. After drug use for 48 h, annexin V/propidium iodide method was used to detect the SO-RB50 cell apoptosis in each group. Results The IC50 value of vincristine treatment on the SO-RB50 cells was 0.11 μmol/L, and ATRT was 12.84 μmol/L. The cell growth curve in control group rose gradually along with the extended culture time, but after vincristine and ATRA treatment, the cell growth curve was smooth and steady. The cell increment was the least in drug combination group and its cell growth curve was the smoothest. There was significant difference in A450 48 h and 72 h after treatment (Fgrouping = 77.316, P < 0.001; Ftime = 86.985, P < 0.001). Compared with control group, A450 value in drug combination group, vincristine group, ATRA group was significant lower (P < 0.001). Compared with control group, the G2/M phase cell proportion in vincristine group was significantly increased, while the G0/G1 phase cell proportion was significantly decreased; the G0/G1 phase cell proportion in ATRA group was significantly increased, while the S phase cell proportion was significantly decreased (FG0/G1 = 85.878, Fs = 56.455, FG2/M = 85.878, P < 0.001). After 48 h, there was significant difference in SO-RB50 cell apoptosis rate among groups (F = 11.312, P < 0.05). The apoptosis rate in drug combination group was significantly higher than that of other groups (P < 0.001). Conclusions ATRA can increase the sensitivity of SO-RB50 cells to vincristine. Vincristine combined with ATRA treatment can significantly increase the inhibiting effect on SO-RB50 cells, which may be related with promoting cell apoptosis and involving in cell cycle control.

288-292

Authors: Jun Lu, You-Ting Ju, Chang Li, Fu-Zhou Hua, Guo-Hai Xu, Yan-Hui Hu

Objective To study the effects of Transient receptor potential cation channel, subfamily V, member 1 (TRPV1) combined with lidocaine on status and apoptosis of U87-MG glioma cell line, and explore whether local anesthetic produces neurotoxicity by TRPV1. Methods U87-MG cells were divided into control group, gene silencing group, empty vector group and TRPV gene up-regulation group. For cells in each group, flow cytometry was employed to detect the intracellular calcium ion concentration and mitochondrial membrane potential at different time point from cellular perspective. Cell apoptosis of U87-MG was assayed by flow cytometry and MTT from a holistic perspective. Results Calcium ion concentration increased along with time. The concentration in TRPV1 gene up-regulation group was significantly higher than those in other groups at each time point (P < 0.05). After adding lidocaine, mitochondrial membrane potential in U87-MG significantly increased (P < 0.05). This increasing trend in TRPV1 gene up-regulation group was more significant than other groups (P < 0.05), while in TRPV1 gene silencing group, the trend significantly decreased (P < 0.05). Flow cytometry result and MTT result both showed that cell apoptosis in each group significantly increased after lidocaine was added (P < 0.05). This increasing trend in TRPV1 gene up-regulation group was more significant than other groups (P < 0.05), while in TRPV1 gene silencing group, the trend significantly decreased (P < 0.05). Moreover, apoptosis was more severe along with the increasing concentration of lidocaine (P < 0.05). Conclusions In this study, it was proved that lidocaine could dose-dependently induce the increase of intracellular calcium ion concentration, mitochondrial membrane potential and apoptosis in U87-MG glioma cell line. The up-regulation of TRPV1 enhanced cytotoxicity of lidocaine, which revealed the correlations between them. Lidocaine might have increased intracellular calcium ion concentration by activating TRPV1 gene and induced apoptosis of U87-GM glioma cell line by up-regulating mitochondrial membrane potential.

298-301

Authors: Xiao-Mei Guan, Yong-Xin Li, Hai Xin, Jun Li, Zong-Gang Zhao, Yue-Wei Wang, Hao-Fu Wang

Objective To observe the effect of miR-467b on the atherosclerosis (AS) of rats with apolipoprotein E (ApoE) gene knockout (ApoE−/−). Methods ApoE−/− rats were fed with high fat and high cholesterol diet and were randomly divided into group A, group B and group C, with 10 rats in each group. Group A: rats were injected with ApoE agonist through the caudal vein; group B: rats were injected with ApoE antagonist through the caudal vein; group C: as negative control group. Enzyme oxidation method was used to detect the blood lipid levels of rats. Western blotting method was used to detect the aortic lipoprotein lipase (LPL) expression levels of rats. HE staining and oil red O staining were performed to observe the AS lesions and lipid accumulation state. Results Compared with group C, blood lipid level, aortic intima and aortic sinus lipid accumulation area ratio, aortic sinus lesion area and LPL expression level in group A significantly reduced; while blood lipid level, aortic intima and aortic sinus lipid accumulation area ratio, aortic sinus lesion area, and LPL expression level in group B significantly increased, with the statistical difference (P < 0.05). Conclusions miR-467b can alleviate the AS lesions of ApoE−/− rats, and its inhibiting effect on AS may be related to LPL expression.