Acute Developmental Toxicity of Panax notoginseng in Zebrafish Larvae / 中国结合医学杂志

OBJECTIVE@#To evaluate toxicity of raw extract of Panax notoginseng (rPN) and decocted extract of PN (dPN) by a toxicological assay using zebrafish larvae, and explore the mechanism by RNA sequencing assay.@*METHODS@#Zebrafish larvae was used to evaluate acute toxicity of PN in two forms: rPN and dPN. Three doses (0.5, 1.5, and 5.0 µ g/mL) of dPN were used to treat zebrafishes for evaluating the developmental toxicity. Behavior abnormalities, body weight, body length and number of vertebral roots were used as specific phenotypic endpoints. RNA sequencing (RNA-seq) assay was applied to clarify the mechanism of acute toxicity, followed by real time PCR (qPCR) for verification. High performance liquid chromatography analysis was performed to determine the chemoprofile of this herb.@*RESULTS@#The acute toxicity result showed that rPN exerted higher acute toxicity than dPN in inducing death of larval zebrafishes (P<0.01). After daily oral intake for 21 days, dPN at doses of 0.5, 1.5 and 5.0 µ g/mL decreased the body weight, body length, and vertebral number of larval zebrafishes, indicating developmental toxicity of dPN. No other adverse outcome was observed during the experimental period. RNA-seq data revealed 38 genes differentially expressed in dPN-treated zebrafishes, of which carboxypeptidase A1 (cpa1) and opioid growth factor receptor-like 2 (ogfrl2) were identified as functional genes in regulating body development of zebrafishes. qPCR data showed that dPN significantly down-regulated the mRNA expressions of cpa1 and ogfrl2 (both P<0.01), verifying cpa1 and ogfrl2 as target genes for dPN.@*CONCLUSION@#This report uncovers the developmental toxicity of dPN, suggesting potential risk of its clinical application in children.

Extraction of protein concentrate from red bean (Phaseolus vulgaris L.): antioxidant activity and inhibition of lipid peroxidation

Red Bean Protein Concentrate (RBPC) and their hydrolysates were used to evaluate the antioxidant capacity. TheRBPC protein content was in the range of 57.38%–72.68% of the total sample content. RBPC protein profile showeda range of 15–100 kDa. Phaseolin protein was identified with bands of 45 and 50 kDa. Phaseolin protein was foundin all the RBPC samples at the different pHs assayed. In the gastric digestion phase, bands from 60 to 100 kDa weretotally hydrolyzed with pepsin. Phaseolin protein (45 and 50 kDa) presented resistance to gastric hydrolysis. All theRBPCs and gastrointestinal digest presented antioxidant activity using ferric-reducing antioxidant power (FRAP),2,2-azinobis (3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS), oxygen radical absorbance capacity (ORAC), andthiobarbituric acid reactive substances using the in vitro and in vivo methods. RBPC at pH 7.0 presented a value of95.80 µmoL TE/g of RBPC (FRAP); 257.12 µmoL TE/g of RBPC (ABTS), and 1960 µmoL TE/g of RBPC (ORAC).Duodenal digest of RBPC presented high antioxidant activity with 225.77 µmoL TE/g of digest (FRAP); 345.21 µmoLTE/g of digest (ABTS); and 3256 µmoL TE/g of digest (ORAC). Gastric and duodenal digest of RBPC were usedto inhibit lipid peroxidation using the in vitro method presenting a value of 87.95% and 93.0%, respectively. Whenthe in vivo method in zebrafish larvae was used, values were 79.03% and 86.76%, respectively. RBPCs showed noreactive oxygen species (ROS) inhibition. However, RBPCs with gastric and gastrointestinal digests, presented ROSinhibition, 75.30% for gastric digests and 66.40% for gastrointestinal digests.