| ObjectiveGuanghuoxiang, a famous traditional Chinese materia medica, has been widely used in prescription drug for dispelling dampness and curing gastrointestinal disease with the history of more than1,000years. To date, it have been also used as the main compositions in more than40kinds of traditional Chinese medicine preparations. Additional uses of this material include making herbal tea and cooking medical soup to deal with the muggy and moisture weather in daily life. According to the market reports, the amounts of annual production and consumption of this material are relatively large. Traditionally, the medical origins of Guanghuoxiang are the dry stem and leaf of Pogostemon cablin (P. cablin), cutting and removing the underground part. Considering that the underground part contributes large proportions of P. cablin in morphology, it would be of great value to utilize the underground part, including minimizing the processing costs of Guanghuoxiang and maximizing the resource utilization of this plant. However, to date there exists no pharmacological research of the underground part of P. cablin and it remains unclear whether the underground part possess potential utilization values. Therefore, this work aimed to analyzing the chemical constituents and examining the pharmacological effects of the ethanol extract from the underground part of P. cablin (ERP) for the further development and utilization of P. cablin.Methods1. Preliminary analysis of chemical constituent of ERP The chromatographic data of main ingredients in ERP were separated and collected via UPLC-PAD analysis system. The authentications and identifications of main ingredients were based on the retention time and the ultraviolet (UV)-absorption (190to800nm) of the known standard compounds. And the contents of identified compounds were quantitative analyses with peak areas under the standard curves.2. Preliminary evaluation of anti-inflammatory property of ERPKM mice was with pretreatment with ERP (120,240and480mg/kg, p. o.) for7days (q. d.).1h after the last administration, the xylene-induced mouse ear edema model was duplicated, the weight difference of ears was measured and the degree of ear edema and the inhibition rate were evaluated.1h after the last administration, the acetic-acid-induced vascular permeability enhancement model was duplicated, the0D difference of peritoneal lavage fluids was measured and the degree of vascular permeability enhancement and the inhibition rate were evaluated.1h after the last administration, the carrageenan-induced mouse paw edema model was duplicated, the volume difference of mouse paw was measured and evaluating the degree of edema and the inhibition rate were evaluated.3. Preliminary elucidation of anti-inflammatory mechanism of ERPKM mice was with pretreatment with ERP (120,240and480mg/kg for mouse, p. o.) for7days (q. d.).1h after the last administration, the carrageenan-induced mouse paw edema model was duplicated.4h after the carrageenan challenge, mouse was sacrificed. The paw was collected for the analysis of MPO, MDA, TNF-a, IL-1β, IL-6, NO, PGE2, iNOS and COX-2as well as the histopathological evaluation. The liver tissue was collected for the analysis of SOD, GPx and GRd.4. Preliminary study of acute toxicity of ERPThe acute toxicity study was performed via combining median lethal (LD50) experiments and maximum tolerance dose (MTD) tests for a single dose toxicity study.Results1. Data of HPLC-PAD analysis profile indicated that3main ingredients, verbascoside, rosmarinic acid and pogostone were indentified in ERP, and their relative contents were listed as pogostone (more than45mg/g)> rosmarinic acid (more than45mg/g)> verbascoside(more than45mg/g). 2. Data of anti-inflammatory property evaluation indicated that oral administration of ERP (120,240and480mg/kg) significantly attenuated xylene-induced ear edema, decreased acetic acid-induced capillary permeability, and reduced carrageenan-induced paw edema, in a dose-dependent manner.3. Data of preliminary mechanism elucidation demonstrated that ERP abated inflammatory responses in histopathologic examination, suppressed the activities of MPO, COX-2and iNOS, attenuated the levels of TNF-α, IL-1β, IL-6, NO and PGE2, up-regulated the activities of SOD, GPx and GRd.4. Data of preliminary acute toxicity indicated that the LD50was uncountable and the MTD was more than4g/kg for a single dose toxicity study.ConclusionIn summary, this paper analyzed the chemical composition of ERP, and three known compounds:rosmarinic acid, verbascoside and pogostone, were quantified by UPLC-PAD. Preliminary acute toxicity study showed that ERP has a lower toxicity with a relative high safety. Oral administration of ERP possessed considerable anti-inflammatory effects, and its mechanisms may be related to the declining of NO and PGE2via inhibitions of COX-2and iNOS, the down-regulations of pro inflammatory cytokines (TNF-α, IL-1β, IL-6) and the up-regulations of antioxidant enzymes (SOD, GPx and GRd). |
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