Effect of Gecko Extract on Neuroinflammation in Rats with Reserpine-induced Depression via TLR4/NF-κB Pathway / 中国实验方剂学杂志

Objective:To investigate the effects and mechanism of Gecko extract for treatment of depression in rats. Method:The depression rats were induced by intraperitoneal injection of reserpine （0.5 mg·kg^-1）. The successfully modeled rats were randomly divided into model group， fluoxetine group （1.8 mg·kg^-1）， high dose and low dose groups of Gecko extract （12， 6 g·kg^-1）. The rats were given corresponding dose of drugs once a day for 10 days. After administration， the levels of neurotransmitters and inflammatory factors in serum and prefrontal cortex of rats were detected by enzyme-linked immunosorbent assay （ELISA）. The cell changes in hippocampal tissues were observed by hematoxylin-eosin （HE） staining. The mRNA levels of interleukin-6 （IL-6）， nuclear factor-<italic>κ</italic>B （NF-<italic>κ</italic>B）， and tumor necrosis factor （TNF-<italic>α</italic>） in the hippocampus of rats were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The protein levels of Toll-like receptor 4 （TLR4） and NF-<italic>κ</italic>B in hippocampal tissues of rats were detected by Western blot. Result:Compared with the normal group， Gecko extract significantly shortened the immobility time of tail suspension and swimming in mice. Compared with model group， Gecko extract significantly reduced blepharoptosis and retention time in circles for the rats （<italic>P</italic><0.05）， increased the levels of 5-hydroxytryptamine （5-HT） and dopamine （DA） in serum （<italic>P</italic><0.05）， decreased the levels of Monoamine oxidase （MAO）， IL-6， and TNF-<italic>α</italic> in serum （<italic>P</italic><0.05） and prefrontal cortex （<italic>P</italic><0.05）， decreased the mRNA levels of inflammatory cytokines IL-6， NF-<italic>κ</italic>B and TNF-<italic>α</italic> and the protein expressions of TLR4 and NF-<italic>κ</italic>B in the hippocampus of rats （<italic>P</italic><0.05，<italic> P</italic><0.01）， and improved the pathological symptoms of the hippocampus. Conclusion:Gecko extract can significantly alleviate the pathological damage of depression and improve the symptoms of depression， and its mechanism may be due to inhibiting TLR4/NF-<italic>κ</italic>B signaling pathway and reducing the expression of NF-<italic>κ</italic>B， IL-6 and other inflammatory factors in the hippocampus of rats.

DNA barcoding and its utility in commonly-used medicinal snakes / 中国中药杂志

Identification accuracy of traditional Chinese medicine is crucial for the traditional Chinese medicine research, production and application. DNA barcoding based on the mitochondrial gene coding for cytochrome c oxidase subunit I (COI), are more and more used for identification of traditional Chinese medicine. Using universal barcoding primers to sequence, we discussed the feasibility of DNA barcoding method for identification commonly-used medicinal snakes (a total of 109 samples belonging to 19 species 15 genera 6 families). The phylogenetic trees using Neighbor-joining were constructed. The results indicated that the mean content of G + C(46.5%) was lower than that of A + T (53.5%). As calculated by Kimera-2-parameter model, the mean intraspecies genetic distance of Trimeresurus albolabris, Ptyas dhumnades and Lycodon rufozonatus was greater than 2%. Further phylogenetic relationship results suggested that identification of one sample of T. albolabris was erroneous. The identification of some samples of P. dhumnades was also not correct, namely originally P. korros was identified as P. dhumnades. Factors influence on intraspecific genetic distance difference of L. rufozonatus need to be studied further. Therefore, DNA barcoding for identification of medicinal snakes is feasible, and greatly complements the morphological classification method. It is necessary to further study in identification of traditional Chinese medicine.

DNA barcoding of COI gene in some medicinal animals of Lacertilia / 中国中药杂志

To identify some medicinal animals of Lacertilia, in total 59 individuals belonging to 12 species 7 genera 3 families, we used the universal barcoding primers to sequence these species, compared with other homologous sequences (564 bp) obtaining from the GenBank and finally constructed phylogenetic trees using Neighbor-joining, Maximum parsimony and Bayesian inference, respectively. As a result, the mean content of G + C (46.5%) was lower than that of A + T (53.5%). As calculated by Kimera-2-parameter model, the whole individuals mean distance for interspecies and intraspecies was 35. 5% and 1.7%, respectively. The mean distance for interspecies was 21 times as much as that for intraspecies. The mean distance for intraspecies of Gekko swinhonis, Hemidactylus frenatus and G. gecko was greater than 2%, respectively. Further analyses suggested that geographical groups of the three species might be of different subSpecies, even species. Of course, incorporating morphological characters and other unlinked genetic markers in future studies will offer further insights into the divergence. On the basis of phylogenetic trees constructed by COI, our results indicated that the taxonomy of the category (family, genus, and species) by DNA barcoding is consistent with morphological characters. Therefore, DNA barcoding is a useful tool for both identification and phylogeny of medicinal animals of Lacertilia, particularly for nonprofessor identifying authentication of Chinese crude drugs of these species.