1.Quality Control of Arnebia euchroma inZizhu Ointment
Yefang DING ; Xilin LI ; Guobin LIU ; Dongxia NIU
Chinese Journal of Information on Traditional Chinese Medicine 2015;(1):77-79,80
Objective To establish a method for quality control of Arnebia euchroma inZizhu Ointment.Methods The TLC was used to make qualitative identification to Arnebia euchroma in Zizhu Ointment. The content of shikonin was targeted as the evaluation index and the method of orthogonal design was used to optimize the extraction of shikonin. UV-visible spectrophotometry was used for the determination of shikonin.Results Main spots of TLC were clear, and negative control had no interference. The optimum extraction of shikonin was extracted for three times, 30 min for each time, NaOH volume fraction of 1%, NaOH amount of 50 mL. The absorbance of shikonin showed a good linear relationship in the range of 0.010 2-0.035 7 mg (r2=0.999 7), and the average recovery was 103.27%, RSD=4.17%.Conclusion This method was simple and feasible, and can be used for the quality control and the evaluation ofZizhu Ointment.
2.Simultaneous Determination of Contents of Shikonin and β,β'-dimethylacrylshikonin from Zizhu Ointment by HPLC
Yefang DING ; Xilin LI ; Guobin LIU ; Yuquan ZHANG
Chinese Journal of Information on Traditional Chinese Medicine 2015;22(11):76-78
Objective To establish an HPLC method to simultaneously determine the contents of shikonin andβ,β'-dimethylacrylshikonin inZizhu Ointment.Methods The contents of shikonin andβ,β'-dimethylacrylshikonin inZizhu Ointment were determined by HPLC. The chromatographic conditions were Welch Materials XB-C18 Column (4.6 mm × 250 mm, 18.5μm), with mobile phase of acetonitrile-water-formic acid (75:25:0.05), detection wavelength of 516 nm, temperature of 30℃, flow rate of 1.0 mL/min and the load of 20μL.Results Shikonin showed a good linearity in the range of 0.025 6-0.256μg (r2=0.999 5), and the average recovery rate was 98.12% (RSD=1.92%).β,β'-dimethylacrylshikonin showed a good linearity in the range of 0.174 4-2.616μg (r2=0.999 2), and the average recovery rate was 100.46% (RSD=1.17%).Conclusion This method is simple and accurate, which can be used for the quality control and evaluation of Zizhu Ointment.
3.Yinjia Tablets Improve Infertility in Rats with Chronic Salpingitis by Correcting Peripheral Blood Th17/Treg Balance
HUANG Yefang ; ZHU Boyu ; WANG Yan ; DING Weijun ; HUANG Shujie
Chinese Journal of Modern Applied Pharmacy 2023;40(19):2635-2642
OBJECTIVE To explore whether Yinjia tablets improve infertility in rats with chronic salpingitis by correcting the peripheral blood Th17/Treg balance. METHODS One hundred female Wistar rats, 82 of which were constructed as a rat model of chronic salpingitis. After successful modeling, the rats were randomly divided into model group, levofloxacin group (20 mg·kg-1), low-dose Yinjia tablets group(0.38 g·kg-1·d-1), medium-dose Yinjia tablets group(0.76 g·kg-1·d-1) and high-dose Yinjia tablets group(1.52 g·kg-1·d-1), 16 rats in each group, and other 16 rats were set up as control group. The treatment groups were administered by gavage at different doses, and the model group and the control group were given the same amount of saline(20 mL·kg-1·d-1), once a day. The gavage was stopped for 1 d after 7 d, and 7 d was one cycle, with a total of 3 cycles of gavage. After administration, the rats were observed for conception and blood samples and oviductal tissues were collected. HE staining was used to observe the pathological changes of fallopian tube tissues; transmission electron microscopy was used to observe the ultrastructural changes of fallopian tube tissues; ELISA was used to detect TNF-α and IL-6 in serum; flow cytometry was used to detect the ratio of Treg and Th17 cells in peripheral blood; RT-qPCR and Western blotting were used to detect IL-17, IL-10, Foxp3, RORγt mRNA and protein expression. RESULTS Compared with the control group, the cilia and microvilli in the model group were sparse and the mitochondria were swollen; the TNF-α and IL-6 contents, the proportion of Th17 cells, and the expression of IL-17 and RORγt mRNA and protein in the oviduct tissues were significantly higher, and the fertility rate, IL-10, the proportion of Treg cells, and the expression of Foxp3 mRNA and protein in the rats were significantly lower(P<0.05). Compared with the model group, the high-dose group of Yinjia tablets improved the structure of oviduct of rats; significantly reduced the TNF-α, IL-6 content, Th17 cell proportion and IL-17, RORγt mRNA and protein expression in oviduct tissue of model rats; and significantly increased the conception rate and IL-10, Treg cell proportion and Foxp3 mRNA and protein expression of rats(P<0.05). CONCLUSION Yinjia tablets can effectively regulate Th17/Treg balance, which in turn regulates Treg and Th17 cell-mediated inflammatory responses and improves the conception rate in chronic salpingitis rats.