Asian Pacific Journal of Tropical Medicine

253-259

Authors: Santram Lodhi, Abhay K Singhai

Objective To evaluate wound healing potential of flavonoid fractions of Martynia annua (M. annua) Linn. leaves in diabetic rats on the basis of folkloric information and preliminary study. Methods The flavonoid compound luteolin and apigenin were isolated from dried leaves of plant by column chromatography. The two concentrations (0.2% and 0.5% w/w) of luteolin and flavonoid fraction were selected for topically applied as ointment on diabetic wound. The Povidone Iodine Ointment USP was used as a reference. On 18th days, protein content, hydroxyproline and antioxidants (SOD, CAT and GSH) level in granuloma tissues were determined. Results The results showed that, percent wound contraction were observed significantly (P<0.01) greater in MAF fraction and 0.5% w/w of luteolin treatment groups. Presence of matured collagen fibres and fibroblasts with better angiogenesis were observed in histopathological studies. Conclusions In conclusion, our findings suggest that flavonoid fraction (MAF) and luteolin (0.5% w/w) may have potential benefit in enhancing wound healing in diabetic condition, possibly due to free-radical scavenging activity of plant.

265-272

Authors: Ya-Fei Zhu, Guo-Lan Gao, Sheng-Bo Tang, Zhen-Dong Zhang, Qing-Shui Huang

Objective To investigate effect and possible mechanisms of silencing human WFDC2 (HE4) gene on biological behavior changes as cell proliferation, apoptosis, movement and invasion of human serous ovarian cancer cell line SKOV3. Methods Lentiviral WFDC2 gene sequence of small interfering siRNA was stablely transfected into SKOV3 identified by Q-PCR and western-blot. Obtained SKOV3 stable strains with silenced HE4 were measured by proliferation, apoptosis, migration, and invasion. Results Gene sequencing showed that the oligonucleotides were successfully inserted into the expected site. After silencing HE4 in the SKOV3, proliferation was significantly inhibited (P<0.05). G0/G1 phase was arrested by the cell cycle (P<0.01) and capacity of the migration and invasion decreased significantly (P<0.01). Slight early apoptosis ratio and no change of late apoptosis were found without change of Caspase-3 or Bcl-2 protein. Proteins involed in ERK pathway as phosphorylated protein as p-EGFR, p-ERK decreased and protease protein involved in tissue remoding as matrix metalloproteinases MMP-9, MMP-2 and cathepsin B decreased compared with control group. Conclusions HE4 gene plays an important role in regulating proliferation, apoptosis, migration, invasion of serous ovarian cancer cells by ERK pathway and protease system. Its role in apoptosis needs to be further explored, and it may be a potential target for serous ovarian cancer.

280-284

Authors: Sunil Mistry, KR Dutt, J Jena

Objective To investigate the hepatoprotective potential of Sida cordata (Malvaceae) (S. cordata) in experimental rats to validate its traditional claim. Methods Wister albino rats were divided into 6 groups: Group I served as control; Group II served as hepatotoxic (CCl4 treated) group; Group III, IV and V served as (100, 200 and 400 mg/kg b.w.) S. cordata leaf extract (SCLE) treated groups; Group VI served as positive control (Silymarin) treated group. Liver marker enzymes serum glutamate oxyloacetic transaminase, serum glutamic pyruvic transaminase, pancreatic enzymatic antioxidants superoxide dismutase (SOD), lipid peroxidation, catalase (CAT), reduced glutathione (GSH) were measured and compared along with histopathological studies. Results Obtained results show that the treatment with SCLE significantly (P<0.05-<0.001) and dose-dependently reduced CCl4 induced elevated serum level of hepatic enzymes. Furthermore, SCLE significantly (up to P<0.001) reduced the lipid peroxidation in the liver tissue and restored activities of defence antioxidant enzymes GSH, SOD and CAT towards normal levels, which was confirmed by the histopathological studies. Conclusions The results of this study strongly indicate the protective effect of SCLE against CCl4 induced acute liver toxicity in rats and thereby scientifically support its traditional use.

296-299

Authors: Hong Wang

Objective To investigate the effect of ophiopogon-polysaccharide on adiponectin (APN) in gestational diabetic mellitus rat. Methods The model of gestational diabeties mellitus rat were established with streptozotocin by intraperitoneal injection. Model rats were randomly divided into ophiopogon-polysaccharide 125 mg/kg group, 250 mg/kg group, 500 mg/kg group, GDM group and normal pregnancy group (n=10). Ophiopogon-polysaccharide was given by intragastric administration for 14 days. The fasting blood glucose levels of rats at the 0, 7, 14 day were detected respectively. The serum insulin level and APN in fat tissue and placenta were detected at the 14 day. Results The fasting blood glucose level, serum insulin level and APN mRNA in fat tissue and placenta were significantly decreased in gestational diabetes rats treated by ophiopogon-polysaccharide compared with model group (P < 0.05). Conclusions Ophiopogon-polysaccharide could decrease the fasting blood glucose level and serum insulin level, and improve APN mRNA in fat tissue and placenta.

305-310

Authors: Zhen-Guo Cheng, Guo-Dong Zhang, Peng-Qiang Shi, Bao-Shun Du

Objective To explore the expression of Nrf2/ARE pathway in hindbrain tissue after the traumatic brain injury (TBI) and its anti-oxidative stress effect in the secondary nerve injury. Methods The mice with Nrf2 gene knockout were used for the establishment of brain injury model. The experimental animals were divided into four groups: (Nrf2+/+) sham-operation group, (Nrf2+/+) brain injury group, (Nrf2−/−) sham-operation group and (Nrf2−/−) brain injury group. The specimen 24 h after cerebral trauma was selected. Then RT-PCR method was adopted to detect the expression of Nrf2 mRNA in brain; Western blotting method was adopted to detect the levels of Nrf2, HO-1 and NQO1 proteins in brain; ELISA method was adopted to detect the oxidative stress indicators: protein carbonyls, 4-hydroxy-2-nonenal (4-HNE) and 8-hydroxy-2′-deoxyguanosine (8-OHdG). Results The Nrf2 mRNA and protein of Nrf2−/− mice were not expressed, and the difference of the relative amount of Nrf2 mRNA between Nrf2+/+ TBI group and Nrf2+/+ sham-operation group was not statistically significant (P>0.05); the level of Nrf2 protein in Nrf2+/+ TBI group increased significantly compared with the Nrf2+/+ sham-operation group (P<0.01); in the sham-operation groups, the levels of HO-1 and NQO1 proteins of Nrf2−/− mice decreased obviously compared with the Nrf2+/+ mice (P<0.01); after brain injury, the levels of HO-1 and NQO1 proteins of Nrf2+/+ mice increased obviously compared with the corresponding sham-operation group (P<0.01); the levels of HO-1 and NQO1 proteins of Nrf2−/− mice in TBI group had no obvious change compared with the corresponding sham-operation group (P>0.05); there was only a little amount of expression of protein carbonyls, 4-HNE and 8-OHdG proteins in brain tissues in the Nrf2+/+ and Nrf2−/− sham-operation groups, and the difference was not statistically significant (P>0.05); after brain injury, the three oxidative stress indicators were significantly up-regulated in the Nrf2+/+ and Nrf2−/− groups, and the up-regulation of the latter group was more significant (P<0.01). Conclusions After TBI the Nrf2/ARE pathway is activated and the activity of Nrf2 transcription regulation increases. However, the regulation dose not occur in the gene transcription level and only could increase the Nrf2 protein level, while the mRNA expression level has no obvious change. The nerve cell protective effect of Nrf2/ARE pathway in TBI achieves through inhibiting the oxidative stress injuries.

315-319

Authors: Samiran Bandyopadhyay, Pabitra H Patra, Achintya Mahanti, Dipak K Mondal, Premanshu Dandapat, Subhasis Bandyopadhyay, Indranil Samanta, Chandan Lodh, Asit K Bera, Debasish Bhattacharyya, Mihir Sarkar, Kishore K Baruah

Objective To evaluate the antimicrobial efficacy of berberine, a plant alkaloid. Methods Five multi-drug resistant (MDR) STEC/EPEC and five MDR ETEC isolates from yaks with haemorrhagic diarrhoea were selected for the study. Antibacterial activity of berberine was evaluated by broth dilution and disc diffusion methods. The binding kinetics of berberine to DNA and protein was also enumerated. Results For both categories of enterovirulent Escherichia coli (E. coli) isolates, berberine displayed the antibacterial effect in a dose dependent manner. The MIC50 of berberine chloride for STEC/EPEC isolates varied from 2.07 μM to 3.6 μM with a mean of (2.95 ± 0.33) μM where as for ETEC strains it varied from 1.75 to 1.96 μM with a mean of (1.87 ± 0.03) μM. Berberine bind more tightly with double helix DNA with Bmax and Kd of (24.68±2.62) and (357.8±57.8), respectively. Berberine reacted with protein in comparatively loose manner with Bmax and Kd of (18.9±3.83) and (286.2±113.6), respectively. Conclusions The results indicate clearly that berberine may serve as a good antibacterial against multi drug resistant E. coli.

325-332

Authors: Loganathan Karthik, Gaurav Kumar, Kokati Venkata Bhaskara Rao

Objective To investigate the antioxidant activity of marine actinobacteria. Methods The content of total phenolics, the level of antioxidant potential by DPPH radical scavenging activity, metal chelating activity, FRAP method, β carotene assay and NO scavenging activity in extract were determined. Results In all the methods the extract exhibited good scavenging activity except NO scavenging activity. The IC50 values of marine actinobacteria extract on DPPH radical were found to be 41.09 μg/mL. The zone of color retention was 12 mm in β-carotene bleaching assay. DNA protective efficiency of the extracts was also studied using UV-photolysed H2O2-driven oxidative damage to pBR322. HPLC analysis identified some of the major phenolic compounds in extracts, which might be responsible for the antioxidant potential and cyto-protection. It showed a 100% cytotoxic effect in brine shrimp lethality assay within 10 mins. The novel actinobacteria was identified as Streptomyces LK-3 (JF710608) through 16S rDNA Sequencing. Conclusions The results obtained suggest that the extracts bear anti-cancer metabolites and could be considered as a potential source for anti-cancer drug development.