1.Content Determination of 7 Constituents in Wikstroemia indica and Its Principal Component and Cluster Analysis
Lili JIA ; Lan WEI ; Jian ZHAO ; Yudi JIA ; Yu QIU ; Chaoying DU ; Lixin SUN
China Pharmacy 2017;28(33):4706-4710
OBJECTIVE:To establish a method for simultaneous determination of 7 constituents in Wikstroemia indica,and to conduct principal component analysis and cluster analysis. METHODS:HPLC method was adopted. The determination was per-formed on Diamonsil Platisil ODS column with mobile phase consisted of acetonitrile-0.15% triethylamine solution(pH adjusted to 6.0 with phosphoric acid,gradient elution)at the flow rate of 1.0 mL/min. The detection wavelength was set at 280 nm,and col-umn temperature was 40 ℃. The sample size was 10 μL. The principal component analysis and cluster analysis were conducted for the results of content determination by SPSS 22.0 statistical software. RESULTS:The linear ranges were 2.688-53.76μg/mL for nar-ingin(r=0.9998),5.052-101.00 μg/mL for myricetin(r=0.9999),2.052-41.04 μg/mL for arctiin(r=0.9999),2.108-42.16 μg/mL(r=0.9999),5.112-102.20 μg/mL(r=0.9999),0.820-16.42 μg/mL(r=0.9999),2.070-41.40 μg/mL(r=0.9999),respec-tively. The limits of quantitation were no higher than 1.0720 μg/mL,the limits of detection were no higher than 0.3318 μg/mL. RSDs of precision,stability and reproducibility tests were all lower than 2.0%. The recoveries were 97.8%-102.5%(RSD=1.8%, n=6),97.2%-102.0%(RSD=2.0%,n=6),95.2%-100.1%(RSD=1.7%,n=6),95.2%-99.3%(RSD=1.6%,n=6), 97.0%-100.8%(RSD=1.3%,n=6),95.5%-98.6%(RSD=1.1%,n=6),95.0%-99.3%(RSD=1.8%,n=6),respectively. Three main components were belong to the samples of 10 batches of medicinal materials. The samples of medicinal materials from 10 pro-ducing area could be divided into 2 categories. The quality of W. indica from Qingyuan Guangdong and Guiyang Guizhou were bet-ter than others. CONCLUSIONS:The method is simple,precise,stable and reproducible,and it can be used for simultaneous de-termination of 7 constituents in medicinal material. The quality of W. indica from different regions are quite different.
2.Effect of Faeces Bombycis Extract on Cajal Interstitial Cells in Diabetic Gastroparesis Rats Based on PI3K/Akt/mTOR Signaling Pathway
Huizhen ZHANG ; Hong ZHANG ; Wei LI ; Kai ZHAO ; Yudi QIU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(8):66-73
ObjectiveTo explore the therapeutic mechanism of Faeces Bombycis on diabetic gastroparesis (DGP) rats based on phosphatidylinositol 3-kinase/protein kinase B/mammalian rapamycin target protein (PI3K/Akt/mTOR) signaling pathway. MethodDGP rat model was prepared by random selection of 15 out of 105 rats as blank group. The rats successfully constructed were randomly divided into model group, high-,medium- and low- dose groups (3.2, 1.6, 0.8 g·kg-1) and moxapride group (1.5 mg·kg-1), with 12 rats in each group, and were given gavage for 4 weeks. The gastric emptying rate and random blood glucose were measured. The morphological changes of gastric antrum were observed by hematoxylin-eosin (HE) staining, and the expression of the c-Kit gene was analyzed by immunohistochemistry. The apoptosis of Cajal interstitial cells was observed by in situ end labeling (TUNEL) staining, and the protein expressions of PI3K, phosphorylation(p)-PI3K, Akt, p-Akt, mTOR, and p-mTOR were detected by Western blot. ResultCompared with the blank group, the gastric emptying rate of the model group decreased significantly (P<0.01), and the glandular structure of the gastric antrum was destroyed. The expression of c-Kit decreased (P<0.01), and the apoptosis of Cajal interstitial cells (ICC) increased. Compared with the model group, the gastric emptying rate in the high, middle, and low-dose groups of Faeces Bombycis extract and mosapride group increased significantly (P<0.01). The glandular structure of the gastric antrum became closer, and the apoptosis of ICC decreased. The expression of c-Kit in the high dose group of Faeces Bombycis extract increased significantly. After Western blot testing, compared with the blank group, the protein expression of p-Akt/Akt, p-PI3K/PI3K, and p-mTOR/mTOR in the model group increased. Compared with the model group, the protein expression of p-Akt/Akt in the high dose group of Faeces Bombycis extract decreased (P<0.01), and the protein expression of p-PI3K/PI3K decreased in the middle and low dose groups of Faeces Bombycis extract and mosapride group decreased (P<0.05, P<0.01). The protein expression of p-mTOR/mTOR decreased in the low dose group of Faeces Bombycis extract (P<0.05). In terms of random blood glucose, compared with the blank group, the random blood glucose in the model group increased significantly (P<0.01), and compared with the model group, the random blood glucose in the high and middle dose groups of Faeces Bombycis extract decreased significantly (P<0.05). Compared with mosapride group, the protein expression of p-Akt/Akt decreased in the high dose group of Faeces Bombycis extract (P<0.05), and the protein expression of p-PI3K/PI3K increased in the high, middle, and low dose groups of Faeces Bombycis extract (P<0.05, P<0.01). ConclusionFaeces Bombycis extract can increase gastric emptying rate, reduce ICC apoptosis, and lower random blood glucose in DGP rats. The high dose group of Faeces Bombycis extract has a significant effect on inhibiting ICC apoptosis, and its mechanism may be related to the regulation of the PI3K/Akt/mTOR signaling pathway.