DENND1A desensitizes granulosa cells to FSH by arresting intracellular FSHR transportation.

Polycystic ovary syndrome (PCOS) is a complex disorder. Genome-wide association studies (GWAS) have identified several genes associated with this condition, including DENND1A. DENND1A encodes a clathrin-binding protein that functions as a guanine nucleotide exchange factor involved in vesicular transport. However, the specific role of DENND1A in reproductive hormone abnormalities and follicle development disorders in PCOS remain poorly understood. In this study, we investigated DENND1A expression in ovarian granulosa cells (GCs) from PCOS patients and its correlation with hormones. Our results revealed an upregulation of DENND1A expression in GCs from PCOS cases, which was positively correlated with testosterone levels. To further explore the functional implications of DENND1A, we generated a transgenic mouse model overexpressing Dennd1a (TG mice). These TG mice exhibited subfertility, irregular estrous cycles, and increased testosterone production following PMSG stimulation. Additionally, the TG mice displayed diminished responsiveness to FSH, characterized by smaller ovary size, less well-developed follicles, and abnormal expressions of FSH-priming genes. Mechanistically, we found that Dennd1a overexpression disrupted the intracellular trafficking of follicle stimulating hormone receptor (FSHR), promoting its internalization and inhibiting recycling. These findings shed light on the reproductive role of DENND1A and uncover the underlying mechanisms, thereby contributing valuable insights into the pathogenesis of PCOS and providing potential avenues for drug design in PCOS treatment.

Protective Effects of Chaihu Shugan San () on Nonalcoholic Fatty Liver Disease in Rats with Insulin Resistance.

OBJECTIVE: To investigate the protective effects of Chinese medicine formulation Chaihu Shugan San (, CHSGS) on nonalcoholic fatty liver disease (NAFLD) in rats with insulin resistance (IR) and its molecular mechanisms. METHODS: Male Sprague-Dawley rats were randomly divided into six groups: the control group, the model group, Dongbao Gantai group (, DBGT, 0.09 g methionine/kg), CHSGS high-dose group (CHSG-H, 12.6 g crude drug/kg), CHSGS medium-dose group (CHSG-M, 6.3 g crude drug/kg), and CHSGS low-dose group (CHSG-L, 3.15 g crude drug/kg). After establishing the NAFLD rat model and treatment for 8 weeks, total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), free fatty acid (FFA), fasting blood glucose (FBG), fasting insulin (FINS) contents in blood serum, and TC, TG contents in the hepatic homogenate were measured by an automatic biochemical analyzer, and a homeostasis model assessment was applied to assess the status of IR, insulin sensitivity index (ISI), and homeostasis model assessment for insulin secretion (HOMA-IS). The expression levels of adiponectin and leptin mRNA in liver tissue were analyzed by reverse transcription polymerase chain reaction. Pathological changes of livers were observed by hematoxylin-eosin staining of paraffin section. RESULTS: Compared with the model group, the serum levels of TC, TG, FFA, FBG, FINS, IRI, ISI, and the liver levels of TC and TG in CHSG-H, CHSG-M, CHSG-L groups showed significant declines (P<0.01 or P<0.05); the serum levels of HDL-C, HOMA-IS were significantly increased (P<0.01 or P<0.05); the expression of leptin mRNA was dramatically decreased and the expression of adiponectin mRNA was increased in the hepatic tissue (P<0.01 or P<0.05). The fatty deposition of liver cells could also be alleviated. CONCLUSION: CHSGS could up-regulate the expression of adiponectin mRNA and down-regulate the expression of leptin mRNA on the liver, suggesting the CHSGS had positive therapeutic effect on NAFLD in rats with IR.

Anti-myocardial Ischemia Effect and Components of Litchi Pericarp Extracts.

Litchi (Litchi chinensis Sonn.) is a famous fruit in south China, and it is also effective for chest tightness or chest pain, irritability, flatulence, epigastric pain and neuralgic pain, hernia pain and testicular swelling, cough, etc. It is valued because a great amount of polyphenol was found in litchi pericarp. In this paper, we got litchi pericarp pure extract by a simple purification method, then evaluated its activity to clear oxygen free radicals in vitro, and evaluated its myocardial protection effect in vivo through acute myocardial ischemia rat model. The results showed that the pure extract had protective effect on myocardial ischemia injury in a certain dose-effect relationship, which reflected in the electrocardiogram, myocardial pathological morphology and other indicators such as cardiac function enzymes, serum and myocardial antioxidant capacity, and eNOS, Bcl-2 and Bax gene expression. Furthermore, we analyzed the components of pure extract by UPLC-MS, ESI-MS and NMR. The main components of PLPE were procyanidin which were identified as procyanidin B2(1), (-)-epicatechin(2), epicatechin-(4ߠ→ 8,2ߠ→ O â†’ 7)-epicatechin-(4ߠ→ 8)-epicatechin(3), A-type procyanidin trimer(4), B-type procyanidin dimer(5) and procyanidin A2(6).This study proved that litchi pericarp extract may have antioxidant activity and cardioprotection effect. It suggested that litchi pericarp may be good for cardiovascular disease. Copyright © 2017 John Wiley & Sons, Ltd.

[Study on antifungal susceptibility of different extract of Dryopteris fragrans].

OBJECTIVE: To examine the antifungal effect of different extract of Dryopteris fragrans (L.) Schott. in vitro, and screen the effective fraction from those extracts. METHODS: Separated the Dryopteris fragrans extract and got four parts by refluxing extraction,and determined the contents of total phloroglucinol. Disc agar diffusion method and solid agar dilution method were used to determine inhibitory effect. Minimum inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) of different parts of Dryopteris fragrans extract against four strains of common clinical dermatophytes were investigated. RESULTS: The data showed that the contents sequence of total phloroglucinol was in the following order: 95% -ethanol extract > water extract > diethyl ether extract > petroleum ether extract, and the antimicrobial activities against the four dermatophytes were as following order: 95% -ethanol extract > water extract > di-ethyl ether extract > petroleum ether extract. CONCLUSION: The contents of total phloroglucinol in 95% -ethanol extract of Dryopteris fragrans is the highest, and the antifungal activity against dermatophytes in vitro is the strongest. The effective fraction of Dryopteris fragrans is the 95%-ethanol extract.

[Effect of Huangqi gegen decoction (HGD) on TGF-beta1/Smad3 pathway in diabetic cardiomyopathy rats].

OBJECTIVE: To study the effect of HGD on diabetic cardiomyopathy and its mechanism. METHODS: The T2-DM rats model was established by combining high fat diet with STZ. The blood glucose, insulin, myocardial fibrosis and TGF-beta1/Smad3 signaling pathway were observed; TGF-beta1 and Smad3 mRNA expression were detected by RT-PCR method, protein expression detected by immunohistochemical method. RESULTS: HGD obviously reduced fasting blood glucose, insulin, improved insulin resistance, reduced myocardial hydroxyproline contents, lowered cardiac index, significantly inhibited over-expression of TGF-beta1/SMAD3 mRNA and protein in diabetic rats cardiac. CONCLUSION: HGD can obviously prevent experimental diabetic myocardial fibrosis through the regulation effect on TGFbeta1/Smad3 signaling pathway.

[The effect of fufang zhenzhu tiaozhi capsule on lipid metabolism in atherosclerotic rabbits].

OBJECTIVE: To observe the effect of fufang zhenzhu tiaozhi capsule (FTZ) on lipid metabolism in atherosclerotic rabbits. METHODS: 50 rabbits were divided into five groups: normal control group, model group, simvastatin group, the high-dose group and low-dose group of FTZ. The atherosclerotic model was established by feeding the rabbits with high fat diet. The medicine interfered groups were administrated with homologous medicine on the base of high fat diet. All the rabbits were fed for 12 weeks. The blood lipids levels were determined just before the experiment and at the forth, the eighth and the twelfth weekend of the experiment. At the end of the experiment, serum apolipoprotein levels, liver lipid contents and the area of atherosclerotic plaque were determined respectively. RESULTS: Compared with model group, TC, TG, LDL-C and ApoB levels were significantly decreased in FTZ groups (P < 0.05 or P < 0.01). The level of HDL-C and the ratio of ApoA I /ApoB were raised significantly (P < 0.05). The contents of TC,TG in liver tissue and the area percentage of atherosclerotic plaque were also markedly reduced (P < 0.05 or P < 0.01) in the FTZ groups. CONCLUSION: FTZ could efficiently regulate the abnormality of lipid metabolism in experimental atherosclerotic rabbits, and this maybe one mamechanisms of its antiatherogenic effect.

[Simulation study of anti-hypoxia active fraction of Xiangdan injection by support vector machine].

OBJECTIVE: To investigate the anti-hypoxia active fraction of Xiangdan injection. METHODS: The anti-hypoxia experiment under normal pressure with mice was adopted on the basis of orthogonal design, and a support vector machine model was established for pharmacodynamics prediction of TCM prescriptions. Moreover, the anti-hypoxia active fraction of Xiangdan injection was determined by experimental confirmation. RESULTS: The prediction model established in this study could predict the drug actions of different formulas. CONCLUSION: As indicated in model prediction and experimental confirmation, the pharmacodynamic actions of several formulas are superior to that of the original formula, and ADE has the best effect.