1.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.
2.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.

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