1.Reproductive toxicity study of rhubarb extract on early pregnancy mice and its preliminary mechanism
Haifeng WANG ; Bing GUO ; Xuping MA ; Ruichen XUE
Chinese Journal of Immunology 2016;(2):184-188
Objective:To explore the abortion effect and mechanism induced by Chinese herbal medicine rhubarb extract.Methods:The pregnant mice at day 3 of gestation were Drenched by Aqueous extract of rhubarb at different doses (7,5,2.5 g/kg)once per day for five days,and mices in control group were oral administration of same volume of saline.All mices at 12 day of gestation were sacrificed.estradiol and progesterone levels in serum were detected by Radioimmunoassay;Content of IFN-γ, IL-2 and TNF-αin uterine lysates were measured by ELISA, macrophages in endometrium were determined by immunohistochemical method, mast cells in endometrium were tested by Toluidine blue staining, respectively.Results: showed as follows:With increasing dose of rhubarb extract,abortion rate and resorption rate were gradually increased and was 100%abortion in 7g/kg treatment groups.Estrogen and progesterone content showed a downward trend,IFN-γ,IL-2 and TNF-αin uterine lysates and the numbers of macrophages and mast cells were significantly higher in mices Drenched by Aqueous extract of rhubarb than in the control.Conclusion: These results show that rhubarb extract not only interfere stability of pregnancy state in pregnant mices because of its diarrhea,but also can directly affect endometrial environment in early embryonic mice,Resulting in abortion.
2.Intervention of Renal Fibrosis with Traditional Chinese Medicine by Regulating Intestinal Microecology Based on "Combination of State and Target"
Xuping LI ; Hailan MA ; Hongbin MA ; Jinhui WEI
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(4):185-191
Renal fibrosis is the main pathological foundation of chronic kidney diseases progressing to end-stage renal diseases. With complex pathogenic factors and prolonged disease course, it threatens the quality of life of patients and brings about heavy financial burden to medical care. In the instance of intestinal flora disturbance, the internal homeostasis is broken, resulting in various "imbalances". The "combination of state and target" endows the syndrome differentiation-based treatment of renal fibrosis with new connotation from the perspective of intestinal flora reconstruction and microbial diversity restoration. In addition, traditional Chinese medicine (TCM)-targeted intervention of intestinal microecology has unique advantages under the principle of "treating different diseases with the same method", which can guide the diagnosis and treatment of renal fibrosis. To be specific, TCM emphasizes macroscopic regulation of state and microscopic targeting. In view of the inflammatory response, accumulation of endotoxin, and excessive deposition of extracellular matrix (ECM) in the process of renal fibrosis, the strategies for treating this disease have been developed, such as alleviating dampness,removing turbid toxin, and relieving deficiency and stasis. Famous prescriptions in ancient books or compound Chinese medicine prescriptions, classical formulas, Chinese medicine monomers, or active components of Chinese medicine target intestinal microecology. Therefore, from the perspective of common pathogenic factors of renal diseases (renal fibrosis) or pathological product-intestinal microecological imbalance, this article combines TCM basic theory with modern medical pathogenesis, and summarizes the research on TCM intervention of renal fibrosis by regulating intestinal microecology and the scientific connotation of renal fibrosis, which is expected to provide ideas and methods for the product development and related preparations and in-depth molecular biological research.
3.Effect of Total Flavone of Litchi Semen on Proliferation, Migration, and Invasion of HepG2 Cells Based on JAK2/STAT3 Signaling Pathway
Minhang LI ; Xiaocong MA ; Yan TANG ; Jingyun LIANG ; Weisheng LUO ; Xuping HUANG
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(22):85-92
ObjectiveTo study the effect of total flavone of Litchi Semen (TFL) on proliferation, apoptosis, migration, and invasion of hepatoma cells HepG2. MethodMethyl thiazolyl tetrazolium colorimetric (MTT) assay was used to detect the effect of different-dose TFL and cisplatin on the proliferation of HepG2 cells. TdT-mediated dUTP nick-end labeling (TUNEL) assay was used to detect the effects of low, medium, and high-dose (70, 140, 210 mg·L-1) of TFL and cisplatin (60 mg·L-1) on the apoptosis of HepG2 cells, thus selecting the optimal dose of TFL for the follow-up experiment. HepG2 cells were divided into a blank group, a TFL group (140 mg·L-1), a TFL+XL019 group (140 mg·L-1 TFL+0.5 μmol·L-1 XL019), and a TFL+TPI-1 group (140 mg·L-1 TFL+1 μmol·L-1 TPI-1). The effect of TFL on migration and invasion of HepG2 cells were examined by wound healing test and Transwell invasion assay, and the effect of TFL on the expression of epithelial-mesenchymal transition (EMT) marker in HepG2 cells were examined by cell immunofluorescence assay. Western blot was used to detect the expression of key proteins in Janus kinase 2(JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathway after the intervention by TFL. ResultMTT assay showed that the proliferation of HepG2 cells was significantly inhibited by TFL and cisplatin at 24 and 48 h as compared with blank group (P<0.01), and the half maximal inhibitory concentration (IC50) of TFL on HepG2 cells was (136.7±2.40) mg·L-1 at 24 h and (106.8±1.11) mg·L-1 at 48 h. The IC50 of cisplatin on HepG2 cells was (58.48±2.04) mg·L-1 at 24 h and (5.15±0.56) mg·L-1 at 48 h. The results of TUNEL assay showed that TFL induced apoptosis of HepG2 cells. The optimal dose of TFL was 140 mg·L-1. The results of wound healing test showed that compared with the blank group, the TFL group, TFL+XL019 group, and the TFL+TPI-1 group significantly inhibited the migration of HepG2 cells (P<0.05, P<0.01). As compared with the TFL group, the inhibitory effect of the TFL+XL019 Group was significantly increased (P<0.05), while that of the TFL+TPI-1 group was significantly decreased (P<0.01). The Transwell invasion assay showed that compared with the blank group, the TFL group, TFL+XL019 group, and the TFL+TPI-1 group significantly inhibited the invasion of HepG2 cells (P<0.01). As compared with the TFL group, the inhibitory effect of the TFL+XL019 group was significantly increased (P<0.05), while that of the TFL+TPI-1 group was significantly decreased (P<0.01). The results of immunofluorescence showed the intervention of TFL up-regulated the expression of E-cadherin, and down-regulated the expression of Vimentin in HepG2 cells, which was stronger in the TFL+XL019 group and weaker in the TFL+TPI-1 group. The results of Western blot showed that compared with the blank group, the TFL group, TFL+XL019 group, and the TFL+TPI-1 group did not affect the expression of JAK2 or STAT3 protein, but significantly decreased the expression levels of phosphorylatied (p)-JAK2 and p-STAT3 (P<0.05, P<0.01). As compared with the TFL group, the expression levels of p-JAK2 and p-STAT3 in the TFL+XL019 group were significantly decreased (P<0.01), while those in the TFL+TPI-1 group were significantly increased (P<0.01). Compared with the blank group, the TFL group significantly increased the expression level of Src-homology domain 2 containing protein tyrosine phosphatase-1(SHP-1) with sh2 domain (P<0.01). ConclusionTFL has the effects of inhibiting the proliferation, promoting apoptosis of HepG2 cells, and reversing the EMT process of HepG2 cells to reduce the migration and invasion, which are presumably related to the activation of SHP-1 by TFL to block JAK/STAT3 signaling pathway.
4. Diagnostic value of serum miRNA let-7a for laryngeal carcinoma and effects of let-7a on proliferation and apoptosis of laryngeal carcinoma cells
En ZHOU ; Lu LUO ; Lijuan MA ; Juan YIN ; Zhiqiang TAN ; Gangyong MIAO ; Jie LIU ; Xuping XIAO
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2019;54(7):534-539
Objective:
To investigate the diagnostic value of serum miRNAlet-7a in laryngeal carcinoma and the effect of let-7a on proliferation and apoptosis of laryngeal carcinoma cells.
Methods:
Real-time quantitative PCR was used to determine the expression level of serum miRNAlet-7a. The miRNA let-7a mimetic was synthesized and transiently transfected into the laryngeal carcinoma Hep-2 cell line by cationic liposome method. The effects of up-regulation of let-7a expression on laryngeal cancer Hep-2 cells were detected by FCM and MTT assays,respectively. The association of let-7a levels with laryngeal cancer and the diagnostic value for laryngeal cancer were analyzed. Measurement data were taken by