Methylation Characteristics of Perivisceral Fat Gene in Obese Rats with Phlegm-dampness Syndrome and the Effect of Wen Dan Decoction

Objective To observe the characteristics of gene methylation in obese rats with phlegm-dampness syndrome induced by the high-fat diet, and to study the effect of Wen Dan Decoction on gene methylation after the intervention. Methods Methylation sites of genes were detected by the MeDIP-seq method. Bioinformatics method was used to analyze the gene methylation characteristics of obesity with phlegmdampness syndrome and the effect of Wen Dan Decoction. Results (1) There were 3 242 methylation differential loci in dietinduced obesity with phlegm-dampness syndrome, of which 1 243 were down-regulated and 1 999 were up-regulated, involving 1 579 differential genes. GO analysis showed that "offactory receptor activity" and others were enriched. The possible signal pathways involved were "Olfactory transduction""Tuberculosis""Systemic lupus erythematosus" and "Ribosome".(2) After the intervention of Wen Dan Decoction in obesity with phlegmdampness syndrome, 4 046 different methylation loci were obtained, including 1 067 down-regulated loci and 2 979 up-regulated loci, involving 2 068 genes. GO analysis showed that "offactory receptor activity" and others were enriched. These genes involved seven signaling pathways, such as "Metabolic pathways".(3) Between diet-induced obesity with phlegm-dampness syndrome and Wen Dan Decoction intervening obesity with the phlegm-dampness syndrome, 582 common genes of methylation differential genes were obtained. After the intervention of Wen Dan Decoction, the number of GO enrichment items was more than that of obesity with phlegm-dampness syndrome, and even the same GO enrichment items involved more genes. Conclusions The phlegm-dampness syndrome of obesityinduced by diet had the characteristics of gene methylation changes, and the intervention of Wen Dan Decoction could also affect the status of gene methylation. The genes affected by Wen Dan Decoction were closely related to the methylation gene of phlegm-dampness syndrome of obesity-induced by diet but covered a wider range.

Study on the Resolving Phlegm Effect of D-Limonene in Mice with Spleen Deficiency and Phlegm-Dampness Syndrome

Objective:According to Traditional Chinese Medicine theory,spleen deficiency and phlegm-dampness syndrome(SDPDS)are caused by abnormal water metabolism in the body because of spleen dysfunction.The purpose of this article was to evaluate the efficacy of D-limonene(DL)in resolving phlegm in mice with SDPDS from the perspective of regulating the level of aquaporin 3(AQP3).Methods:The model of SDPDS was induced in mice using the multifactor modeling method,which combines internal and external dampness.An artificial climate box was used to create a humid environment,whereas the irregular diet was caused by different feeding methods on odd-even days.The mice were divided into blank control,model group,DL low-dose,DL high-dose,and positive groups.The mice were modeled and treated for 7 day.Levels of gastrin and amylase(AMS)in the serum,mucus secretion in the trachea,and AQP3 in the tissue near the gastric cardia.Results:DL significantly reduced mucus secretion in the trachea(P<0.001).It also increased the level of AMS in the serum(P<0.01)and decreased the level of AQP3 in the gastric tissue(P<0.01).Conclusions:Mice with SDPDS exhibited disturbed water metabolism and significantly increased AQP3 levels.DL can restore the levels of AQP3 and plays an important role in resolving phlegm.This study may also help expand the efficacy of natural drugs containing DL and improve the utilization of natural drug resources.

Effect of Electro-acupuncture on Expression of IRS-1/PI3K/GLUT4 Pathway in Ovarian Granulosa Cells of Infertile Patients with Polycystic Ovary Syndrome-Insulin Resistance of Phlegm-Dampness Syndrome

Objective:To evaluate the effect of electro-acupuncture(EA)in infertile patients with phlegmdampness polycystic ovary syndrome-insulin resistance(PCOS-IR).Methods:Seventy-six PCOS-IR patients who underwnet in vitro fertilization and embryo transfer(IVF-ET)were equally assigned to two groups according to a random digital table:the EA group and the control group,with 38 cases in each group.Before undergoing IVF,the two groups were treated with EA or pseudo-acupuncture,respectively,for 3 menstrual cycles.The intervention was 25 min twice a week until the day of oocyte collection.The selected acupoints were Zhongwan(RN 12),Tianshu(ST 25),Daheng(SP 15),Daimai(GB 26),Qihai(CV 6),Guanyuan(CV 4),and bilateral points including Xuehai(SP 10),Fenglong(ST 40),Zusanli(ST 36),and Yinlingquan(SP 9).Evaluation of phlegm-dampness syndrome score and IR score were carried out before and after treatment.Additionally,the number of oocytes retrieved,transplantable embryo rate,high-quality embryo rate,clinical pregnancy rate and live birth rate were compared between the two groups.Real-time polymerase chain reaction analysis was used to monitor the m RNA expression of the insulin receptor substrate(IRS-1),phosphatidylinositiol 3-kinase(PI3 K)and glucose transport factor 4(GLUT4)in ovarian granulosa cells.Results:EA treatment reduced the phlegm-dampness syndrome score as well as the IR scores compared with the control group(P<0.05).No significant differences in the number of oocytes retrieved and clinical pregnancy rate between the two groups(P>0.05).Moreover,the transplantable embryo rate[49.0%(284/580)vs.41.9%(273/652)],high-quality embryo rate[36.6%(104/284)vs.27.8%(76/273)],and live birth rate[50%(19/38)vs.26.3%(10/38)]in the EA group were significantly higher than in the control group(P<0.05).Gene expression analyses revealed significantly elevated IRS-1,PI3 K and GLUT4 m RNA in ovarian granulosa cells of the EA group compared with the control group(P<0.05).Conclusions:EA may ameliorate the effects of phlegm-dampness syndrome and ovarian IR in PCOS-IR patients.Mechanistically,this effect might be through an upregulation of the IRS-1/PI3 K/GLUT4 signaling pathway,which may result in improved oocyte quality and embryonic development potential.(Registration No.Chi CTR1800015453)