[Spleen deficiency and phlegm dampness syndrome model in rats treated by Citri Reticulatae Pericarpium based on metabolomics].

The present study analyzed the effect of Citri Reticulatae Pericarpium on endogenous metabolites in spleen deficiency and phlegm dampness syndrome by metabolomics, and explored the underlying mechanism of Citri Reticulatae Pericarpium in the treatment of spleen deficiency and phlegm dampness syndrome.The model of spleen deficiency and phlegm dampness syndrome was induced in rats by the multi-factor modeling method.The intervention effects of Citri Reticulatae Pericarpium on rats with spleen deficiency and phlegm dampness syndrome were preliminarily evaluated by observing the pathological changes of rat liver tissues and measuring the plasma content of pathological and biochemical indexes such as triglyceride(TG), total cholesterol(TC), low-density lipoprotein cholesterol(LDL-C), and high-density lipoprotein cholesterol(HDL-C).Immunohistochemistry was used to detect the expression of AQP2 in the kidney, AQP3 in the colon, and AQP5 in the submandibular gland, and the effect of Citri Reticulatae Pericarpium on aquaporin expression in rats with spleen deficiency and phlegm dampness syndrome was evaluated.Furthermore, UHPLC-ESI-MS/MS was used to analyze the metabolic profiles of rat plasma samples.Multiple methods, such as principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA) were used for pattern recognition.Differential metabolites were screened out by t-test and variable importance in projection(VIP), followed by pathway analysis based on MetaboAnalyst 5.0.As revealed by experimental results, Citri Reticulatae Pericarpium could improve the pathological changes of liver tissues, increase the levels of HDL-C in the plasma, reduce the levels of TC, TG, and LDL-C, and enhance the expression of AQP2 in the kidney, AQP3 in the colon, and AQP5 in the submandibular gland of rats with spleen deficiency and phlegm dampness syndrome.In addition, 87 differential metabolites of spleen deficiency and phlegm dampness syndrome were screened out by UHPLC-ESI-MS/MS(the levels of 39 metabolites increased significantly and the levels of 48 metabolites decreased significantly), with the representatives of glycine, L-isoleucine, N-acetyl-L-tyrosine, xanthine, hypoxanthine, and trigonelline.The differential metabolites were mainly enriched in the pathways of steroid hormone biosynthesis, linoleic acid metabolism, and purine metabolism.This study distinguished and revealed the characteristic metabolic pattern of spleen deficiency and phlegm dampness syndrome by metabolomics.The preliminary construction of the OPLS-DA model provides an objective basis for the differentiation of spleen deficiency and phlegm dampness syndrome in traditional Chinese medi-cine(TCM), as well as ideas and methods for exploring the biological basis of TCM syndrome from the molecular level and the overall level.

Regulatory Effect of Modified Erzhu Erchentang on Patients with Polycystic Ovary Syndrome and Spleen Deficiency and Phlegm Dampness / 中国实验方剂学杂志

Objective:To observe the regulatory effect of modified Erzhu Erchentang on metabolization of polycystic ovary syndrome （PCOS） with spleen deficiency and phlegm dampness syndrome. Method:Patients 140 cases were divided into control group and observation group. Both groups were given metformin hydrochloride tablets， 500 mg/time， 3 times/day. Control group was given Yuejun Erchen pills， 0.5 g/time， 3 times/day， while observation group was given modified Erzhu Erchentang， 1 dose/day. The course of treatment lasted for 24 weeks. Before and after treatment， levels of fasting blood glucose （FBG）， fasting insulin （FINS）， glycosylated hemoglobin Alc （HbA1c）， 2-hour postprandial blood glucose （2 h PG）， blood lipid， waist circumference （WC）， body mass index （BMI）， waist hip ratio （WHR）， luteinizing hormone （LH）， follicle stimulating hormone （FSH）， serum testosterone （T）， estradiol （E₂）， dehydroepiandrosterone sulfate （DHEAS）， sex hormone binding globulin （SHBG）， leptin （LP）， adiponectin （APN）， resistin， visfatin and tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>） were detected. Homeostasis model assessment insulin resistance （HOMA-IR） was calculated， modified Erzhu Erchentang was scored， and recovery of menstruation and ovulation and ovarian volume were recorded. Result:Levels of FBG， 2 h PG， HbA1c， FINS， HOMA-IR， triglyceride （TG）， total cholesterol （TC）， low-density lipoprotein cholesterol （LDL-C）， LH， FSH， T， E₂， DHEAS， LP， resistin， visfatin and TNF-<italic>α</italic> in observation group were lower than those in control group （<italic>P</italic><0.01）， and levels of BMI， WC and WHR were lower than those of control group （<italic>P</italic><0.05）. And levels of high-density lipoprotein cholesterol （HDL-C）， SHBG and APN were higher than those in control group （<italic>P</italic><0.01）. Score of modified Erzhu Erchentang was lower than that in control group （<italic>P</italic><0.01）， and ovarian volume was smaller than that in control group （<italic>P</italic><0.01）. The normal rate of BMI was 49.23% （32/65）， which was higher than 30.30% （20/66） in control group （<italic>χ</italic>²=5.151， <italic>P</italic><0.05）. The normal rate of blood lipid was 93.85% （61/65）， which was higher than 81.82 % （54/66） in control group （<italic>χ</italic>²=4.418， <italic>P</italic><0.05）. The normal rate of blood glucose was 96.92% （63/65）， which was higher than 86.36% （57/66） in control group （<italic>χ</italic>²=4.474， <italic>P</italic><0.05）. Conclusion:In addition to adipocytokines， modified Erzhu Erchentang could regulate adipokines of patients of PCOS with spleen deficiency and phlegm dampness， improve glucose， lipid metabolism and overweight， adjust endocrine hormone， reduce clinical symptoms and improve ovarian structure， so as to create conditions for conception.

Effect of Modified Cangfu Daotantang on Metabolism and Pregnancy in Patients with Polycystic Ovary Syndrome / 中国实验方剂学杂志

Objective:To observe the effect of modified Cangfu Daotantang on metabolism and pregnancy in patients with spleen deficiency and phlegm-dampness type polycystic ovary syndrome （PCOS）. Method:One hundred and twelve patients were randomly divided into control group and observation group according to the random number table. Both groups took non-pharmacological interventions， oral metformin hydrochloride， 500mg/time， 3 times/day； oral ethinyl estradiol and cyproterone tablets， 1 tablet/time， 1 time/day， starting from the third to fifth day of menstruation and lasting for twenty-one days， for a total of 3 menstrual cycles. Patients in control group additionally took Erchen pills orally， 10 g/time， 2 times/day， while patients in observation group additionally took modified Cangfu Daotantang orally， 1 dose/day. The course of treatment was six menstrual cycles in both groups （or termination after conception）. The waist-to-hip ratio （WHR）， body mass index （BMI）， insulin resistance index （HOMA-IR）， pancreatic <italic>β</italic>-cell function （HOMA-<italic>β</italic>）， triglycerides （TG）， low-density lipoprotein （LDL） and non-high-density lipoprotein （nHDL） elevation after treatment were compared. The number of ovulation cycles monitored by B-ultrasound （6 menstrual cycles）， ovulation rate， human chorionic gonadotropin （HCG） day endometrial thickness， follicle diameter， cervical mucus score>8 points and endometrial morphology type A rate were measured and recorded. The recovery of menstruation， pregnancy and early miscarriage were recorded. Luteinizing hormone （LH）， estradiol （E₂）， follicle stimulating hormone （FSH）， dehydroepiandrosterone sulfate （DHEAS）， testosterone （T）， anti-Müllerian hormone （AMH） levels， and insulin before and after treatment -Like growth factor-1 （IGF-1）， leptin （LP）， adiponectin （APN）， growth differentiation factor-9 （GDF-9） and tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>） levels were detected. Result:WHR， BMI and HOMA-IR levels of the observation group were lower than those of the control group （<italic>P</italic><0.05， <italic>P</italic><0.01）. HOMA-<italic>β</italic> level was higher than that in the control group （<italic>P</italic><0.01）. The increase rates of LDL， TG， and nHDL in the observation group were 19.61%（10/51），25.49%（13/51），23.53%（12/51）， respectively， lower than 41.18%（21/51），47.06%（24/51），45.10%（23/51）respectively in the control group （<italic>χ</italic>²=5.607， <italic>χ</italic>²=5.131， <italic>χ</italic>²=5.263， <italic>P</italic><0.05）. The menstrual recovery rate in the observation group was 90.20% （46/51）， higher than 72.55% （37/51） in the control group （<italic>χ</italic>²=5.239，<italic>P</italic><0.05）. The observation group had more ovulation cycles than the control group （<italic>P</italic><0.01）. The pregnancy rate in the observation group was 50.98% （26/51）， higher than 31.37% （16/51） in the control group （<italic>χ</italic>²=4.047，<italic>P</italic><0.05）. On HCG day after treatment， the endometrial thickness and follicle diameter in the observation group were better than those in the control group （<italic>P</italic><0.01）. The proportion of patients with cervical mucus score> 8 points was 78.43% （40/51） in the observation group， higher than 56.86% （29/51） in the control group （<italic>χ</italic>²=5.420，<italic>P</italic><0.05）. The intimal morphology type A rate in the observation group was 52.94% （27/51）， higher than 31.37% （16/51） in the control group （<italic>χ</italic>²=4.864，<italic>P</italic><0.05）. The levels of AMH， E₂， DHEAS， LH， T ， IGF-1， LP and TNF-<italic>α</italic> in the observation group were lower than those in the control group （<italic>P</italic><0.01）， while the APN and GDF-9 levels were superior to those in the control group （<italic>P</italic><0.01）. Conclusion:On the basis of conventional western medicine intervention， modified Cangfu Daotantang can regulate abnormal metabolism and reproductive endocrine in patients with PCOS， improve conception， and regulate the expression of IGF-1， GDF-9， adipocytokines and inflammatory factors， improve ovulation and improve pregnancy rate.