The present study aimed to analyze the effects and underlying mechanisms of microRNA (miR)-29-3p on the proliferation and secretory abilities of prolactinoma cells by targeting insulin-like growth factor (IGF)-1/β-catenin. The relationship between miR-29a-3p and the survival of prolactinoma cells was analyzed with the Kaplan-Meier method in reference to The Cancer Genome Atlas. The expression levels of miR-29a-3p and IGF-1 in MMQ and GH3 cells were detected. A dual-luciferase reporter gene assay was performed to verify the combination of miR-29a-3p and IGF-1. Cells were transfected with a miR-29a-3p mimic and/or IGF-1 pcDNA3.1 to analyze the effects on the proliferation, apoptosis and secretion of prolactin (PRL) and growth hormone (GH) of prolactinoma cells. The effects on β-catenin in the cytoplasm and nucleus were investigated by western blot analysis. The results showed that miR-29a-3p expression was low in MMQ and GH3 cells. Overexpression miR-29a-3p inhibited IGF-1 mRNA and protein expression. miR-29a-3p inhibited cell proliferation and PRL and GH expression, and promoted apoptosis by inhibiting IGF-1. Increasing the expression of miR-29a-3p increased β-catenin levels in the cytoplasm, whereas IGF-1 promoted β-catenin activation and entry into the nucleus, and reversed the inhibitory effects of miR-29a-3p on β-catenin. To conclude, miR-29a-3p inhibited the proliferation and secretory abilities of prolactinoma cells by inhibiting nuclear translocation of β-catenin via a molecular mechanism that is inseparable from IGF-1.
Central precocious puberty (CPP) severely affects children’s physical and mental health and needs to be treated promptly and effectively. This article aimed to research the therapeutic effect of Shugan Xiehuo Formula (SXF) on CPP. A female CPP rat model was established and then treated with leuprolide and different doses of SXF. Sex organ volume and index were measured. Ovaries and uteri were visualized by hematoxylin-eosin staining. The concentrations of follicle-stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), and estradiol (E2) in peripheral blood were determined. The expression levels of gonadotropin-releasing hormone (GnRH), gonadotropin-releasing hormone receptor (GnRHR), estrogen receptor alpha (ERα), and G protein-coupled receptor 30 (GPR30) in the hypophysis were investigated by Real-Time Quantitative Reverse Transcription PCR and western blot. GnRH expression in the hypothalamus and GnRHR expression in the ovary were detected by immunohistochemistry. SXF reduced the volume of the bilateral ovaries, as well as the volumes of the uterus, hypothalamus, and hypophysis in the female CPP rats and diminished the index of the ovary, uterus, hypothalamus, and hypophysis in the female CPP rats ( P < 0.05 or P < 0.01 ). SXF treatment inhibited follicle maturation and uterine wall thickening in the female CPP rats. SXF decreased the concentrations of FSH, LH, PRL, and E2 in the peripheral blood in the female CPP rats ( P < 0.01 or P < 0.001 ). SXF suppressed the expressions of GnRH, GnRHR, ERα, and GPR30 in the hypophysis ( P < 0.05 ), the expression of GnRH in the hypothalamus ( P < 0.01 ), and the expression of GnRHR in the ovaries ( P < 0.001 ) of the female CPP rats. Overall, our study revealed that SXF had therapeutic effects on CPP in female rats. This is worthy of promoting clinically.
Laggera pterodonta is a famous Chinese herbal medicine in China, which has the effect of treating viral infection, bronchial and sore throat. Yunnan Province of China is the main production area, and Laggera pterodonta grows in a wide range of areas in Yunnan Province. The climates of the respective growing places are also different, which may be different for the accumulation of medicinal components in it, and the evaluation of different the quality of the traditional Chinese medicine of Laggera pterodonta of origin is of great significance. At present, there was still a lack of reliable technology for the origin identification of Laggera pterodonta, especially the origin of Laggera pterodonta cannot be identified from the appearance, which was very difficult to evaluate the efficacy of Laggera pterodonta from different origins. Therefore, it is of great significance to develop a quick and easy method for the origin identification of Laggera pterodonta. Thirty-four samples of Laggera pterodonta from five main production areas of Dali, Jianshui, Lancang, Simao and Yuanjiang in Yunnan Province were used as the research objects. UPLC and PDA detector were used to collect fingerprints, the common peaks were obtained using the "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2012 Edition)", and then carry out PCA and OPLS-DA analysis by R and SIMCA-P, respectively. Laggera pterodonta from different origins can be distinguished by OPLS-DA, which was consistent with the PCA analysis results. Through the comparison of VIP values, 8 characteristic components with great differences are obtained, and the UV absorption of these characteristic components was mainly concentrated at 320-350 nm and 250-280 nm, possibly characteristic UV absorption of terpenes, sesquiterpenes, and alkaloids. Through the comparison of the standard products, it was confirmed that the two characteristic components were chrysosplenetin and artemisetin, and the quantitative analysis of these two compounds in the Laggera pterodonta from different origins was carried out. The results of variance analysis showed that the areas with the most significant difference in chrysosplenetin content were Lancang and Honghe (p=9.03×10-5), and the areas with the most significant difference in artemisetin content were Lancang and Simao (p=5.10×10-7). The Simao and Dali areas have lower levels of artemisetin than other areas, with the highest levels being the Lancang area. The content of chrysosplenetin in Dali area was lower than that in other areas, and the content was higher in Lancang area. It was speculated that Laggera pterodonta in Lancang area may be better than other origins in terms of quality and efficacy of Chinese medicinal materials. Chrysosplenetin can also be used as one of the important indicators for quality control and origin traceability of Laggera pterodonta. Using UPLC combined with chemometric methods, Laggera pterodonta from different regions of China’s Yunnan Province, can be effectively identified.
Background: Central precocious puberty (CPP) severely affected children’s physical and mental health, which needs to be treated promptly and effectively.Objective: To research the therapeutic effect of Shugan Xiehuo Formula (SXF) on CPP.Methods: CPP female rat model was established and treated by leuprolide and different dose of SXF. Sex organs volume and index were measured. The ovary and uterus was experienced HE staining. Concentration of follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL) and estradiol (E2) in peripheral blood was determined. Expression of gonadotropin-releasing hormone (GnRH), gonadotropin-releasing hormone receptor (GnRHR), estrogen receptor alpha (ERα) and G protein-coupled receptor 30 (GPR30) in hypophysis was investigated by qRT-PCR and Western Blot. GnRH expression in hypothalamus and GnRHR expression in ovary was detected by immunohistochemistry.Results: SXF reduced the volume of bilateral ovaries, uterus, hypothalamus and hypophysis in CPP female rat, and diminished the index of ovary, uterus, hypothalamus and hypophysis in CPP female rat (P < 0.05 or P < 0.01). SXF treatment inhibited follicles maturation and uterine wall thickening in CPP female rat. SXF declined FSH, LH, PRL and E2 concentration in peripheral blood in CPP female rat (P < 0.01or P < 0.001). SXF suppressed the expression of GnRH, GnRHR, ERα and GPR30 in hypophysis, the expression of GnRH in hypothalamus and GnRHR in ovary of CPP female rat (P < 0.05, P < 0.01 or P < 0.001).Conclusions: SXF had effective therapeutic effects on CPP female rat, which was worthy of promotion clinically.
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