Analysis of the chemical compositions of Hirudinaria manillensis

Aim To determine and analyze fatty acids, amino acids and trace elements of Hirudinaria manillensis. Methods Contents of fatty acids, amino acids and trace elements were determined by GC/MS, HPLC and ICP-AES. Results H. manillensis contained sixteen fatty acids, among which the percentage of unsaturated fatty acids is 34.05%. The eighteen amino acids were detected, which included eight essential amino acids in human body with a ratio of 33.53 %. There were eight trace elements （Ca, Cr, Cu, Fe, Mg, Mn, V and Zn） from H. manillensis, and the content of Ca, Zn and Fe were apparently higher than those of other elements. Conclusion These methods are simple and accurate. This study reports routine quantitative methods for the analysis of multiple components in H. manillensis by GC/MS, HPLC and ICP-AES.

热带水蛭Hirudinaria manillensis睡液提取物的体内降糖作用(英文)

The anticoagulant effect of leech saliva was traditionally employed in the treatment of diabetes mellitus complications such as peripheral vascular complications.This study was carried out to examine the effect of leech saliva extract(LSE)on blood glucose levels in alloxan-induced diabetic rats.First,LSE was collected from leeches which were fed on a phagostimulatory solution.Second,total protein concentration was estimated using the Bradford assay.Third,diabetic rats were injected subcutaneously(sc)with LSE at doses of 500 and 1 000μg·kg 1body weight(bw).Other diabetic rats were injected sc with insulin at doses of 10 and 20 U·kg 1bw.Another group was injected simultaneously with LSE(250μg·kg 1bw)and insulin(10 U·kg 1bw).Fasting blood glucose(FBG)concentrations were monitored during a study period of eight hours at regular intervals.Findings showed that both doses of LSE resulted in a significant and gradual decrease in FBG starting from 10%18%downfall after two hours of injection reaching the maximal reduction activity of 58%after eight hours.Remarkably,LSE was sufficient to bring the rats to a near norm-glycemic state.The high dose of insulin induced a severe hypoglycemic condition after 2 4 h of injection.The lower dose was able to decline FBG for 2 6 h in rats which became diabetic again after 8 h.On the other hand,the concurrent injection of low doses of LSE and insulin produced a hypoglycemic effect with all rats showing normal FBG levels.Taken together,these findings indicated that the subcutaneous injection of LSE of the medicinal Malaysian leech was able to provide better glycemic control compared with insulin.Moreover,the synergism between LSE and insulin suggests that LSE could be utilized as an adjuvant medication in order to reduce insulin dosage or to achieve better control of blood glucose.

Identification and characterization of a novel elastase inhibitor from Hirudinaria manillensis

A large number of protease inhibitors have been found from leeches,which are essential in various physiological and biological processes.In the curret study,a novel elastase inhibitor was purified and characterized from the leech of Hirudinaria manillensis,which was named HMEI-A.Primary structure analysis showed that HMEI-A belonged to a new family of proteins.HMEI-A exerted inhibitory effects on elastase and showed potent abilities to inhibit elastase with an inhibition constant(Ki)of 1.69×10^(−8) mol·L^(−1).Further study showed that HMEI-A inhibited the formation of neutrophil extracellular trap(NET).These results suggested that HMEIA from the leech of H.manillensis is a novel elastase inhibitor which can suppress NET formation.It may play a significant role in blood-sucking of leeches and is a potential candidate as an anti-inflammatory agent.

Antibiotic susceptibility of body surface and gut micro flora of two aquatic leech species(Hirudinaria manillensis and Hirudinaria javanica)in Malaysia

Objective:To elucidate the antibiotic susceptibility of body surface and gut associated microflora of two local aquatic leech species Hirudinaria manillensis and Hirudinaria javanica.Methods:Four commercially available antibiotics(doxycycline,chloramphenicol,tetracycline and ciprofloxacin)were used in this study.A total of 13 isolated gut and two surface micro flora from Hirudinaria manillensis and two gut and two surface micro flora from Hirudinaria javanica were tested for their antibiotic susceptibility.Results:Based on the susceptibility,it was observed that all the isolated bacteria were found to be susceptible to at least three of the antibiotics except Microbacterium resistens,Serratiamarcescens and Morganella morganii.This study also found that the bacterial species Bacillus fusiformis has displayed resistance against tetracycline and Tsukamurella inchonensis against chloramphenicol.Conclusions:Among all the antibiotics tested,ciprofloxacin was found to be the best bactericidal agent.The immersion of leeches in ciprofloxacin before the application to the patient may be beneficial to prevent invasive infection of the patient.Further study is needed to sterilize the live leech by immersion/oral mode of administration for the tested antibiotics.