A Novel Glycated Hemoglobin A1c-Lowering Traditional Chinese Medicinal Formula, Identified by Translational Medicine Study

<div><p>Diabetes is a chronic metabolic disorder that has a significant impact on the health care system. The reduction of glycated hemoglobin A1c is highly associated with the improvements of glycemic control and diabetic complications. In this study, we identified a traditional Chinese medicinal formula with a HbA1c-lowering potential from clinical evidences. By surveying 9,973 diabetic patients enrolled in Taiwan Diabetic Care Management Program, we found that Chu-Yeh-Shih-Kao-Tang (CYSKT) significantly reduced HbA1c values in diabetic patients. CYSKT reduced the levels of HbA1c and fasting blood glucose, and stimulated the blood glucose clearance in type 2 diabetic mice. CYSKT affected the expressions of genes associated with insulin signaling pathway, increased the amount of phosphorylated insulin receptor in cells and tissues, and stimulated the translocation of glucose transporter 4. Moreover, CYSKT affected the expressions of genes related to diabetic complications, improved the levels of renal function indexes, and increased the survival rate of diabetic mice. In conclusion, this was a translational medicine study that applied a “bedside-to-bench” approach to identify a novel HbA1c-lowering formula. Our findings suggested that oral administration of CYSKT affected insulin signaling pathway, decreased HbA1c and blood glucose levels, and consequently reduced mortality rate in type 2 diabetic mice.</p></div

Effect of CYSKT on blood glucose levels in normal mice.

<p>Normal mice were administered orally various amounts of CYSKT. Glucose (4 g/kg) was injected intraperitoneally 15 min later, blood samples were collected at intervals, and blood glucose levels were measured by a glucometer (A). (B) AUC of glucose tolerance assay. Values are mean ± SD (<i>n</i> = 5). *<i>p</i><0.05, **<i>p</i><0.01, compared with mock.</p

Effects of CYSKT on renal function indexes and survival rate in type 2 diabetic mice.

<p>(A) CYSKT (200 mg/kg) was orally given to type 2 diabetic mice for 30 consecutive days. The levels of BUN and creatinine in sera were measured by an autoanalyzer. Values are mean ± SD (<i>n</i> = 5). **<i>p</i><0.01, compared with mock. (B) Long-term survival rate. Type 2 diabetic mice were administered orally with 200 mg/kg CYSKT and/or 20 mg/kg TZD for 120 consecutive days. The number of death was observed daily.</p

Effects of CYSKT on IR phosphorylation and GLUT-4 translocation.

<p>(A) Phospho-IR ELISA. Left panel: HepG2 cells were treated with 0.5 µM insulin or various amounts of CYSKT. Ten minutes later, cellular proteins were collected and the levels of phosphorylated IR were measured by ELISA. Values are mean ± SD (<i>n</i> = 6). Right panel: BALC/c mice were given orally with various amounts of CYSKT. One hour later, mice were sacrificed, and livers were collected for analysis. Values are mean ± SD (<i>n</i> = 6). *<i>p</i><0.05, ***<i>p</i><0.001, compared with mock. (B) Phospho-IR ELISA. Type 2 diabetic mice were administered orally 200 mg/kg CYSKT for 30 days. Proteins were extracted from intestines and muscles, and the levels of phosphorylated IR were measured by ELISA. Values are mean ± SD (<i>n</i> = 5). *<i>p</i><0.05, **<i>p</i><0.01, ***<i>p</i><0.001, compared with mock. (C) IHC. Type 2 diabetic mice were administered orally 200 mg/kg CYSKT for 30 consecutive days. Muscle tissues were collected and the sections were stained by IHC using antibody against GLUT-4 (100× and 400× magnification). Photos are representative images (<i>n</i> = 5).</p

DataSheet1_Anti-inflammatory sesquiterpene and triterpene acids from Mesona procumbens Hemsley.PDF

Mesonaprocumbens Hemsley is a plant conventionally processed to provide popular food materials and herbal medicines in Asia. In this study, six triterpene acids, including five new ones (mesonaic acids D-H, 1–5), and one proximadiol-type sesquiterpene (7) were isolated from the methanolic extract of the air-dried M. procumbens. Chemical structures of 1‒7 were established by spectroscopic methods, especially 2D NMR techniques (1H–1H COSY, HSQC, HMBC, and NOESY) and HRESIMS. Concerning their biological activities, compounds 1, 2, 6, and 7 were examined manifesting high inhibition toward the pro-inflammatory NO production with EC50 values ranging from 12.88 to 21.21 µM, outrunning the positive control quercetin (24.12 µM). The mesoeudesmol B (7) identified from M. procumbens is the very first example, which exhibited high anti-inflammatory activity diminishing the level of the lipopolysaccharide-induced NO in RAW264.7 macrophage cells, thereby suppressing the secretion of pro-inflammatory cytokines TNF-α and IL-6 and the level of two critical downstream inflammatory mediators iNOS and COX-2.</p

Effects of CYSKT on the levels of HbA1c and blood glucose in type 2 diabetic mice.

<p>Type 2 diabetic mice were administered orally 200 mg/kg CYSKT for 30 consecutive days. Blood samples were collected every 10 days and measured for HbA1c levels (A) and fasting blood glucose levels (B). Glucose tolerance test was performed on 30th day after CYSKT administration. Mice were fasted for 4 h, glucose (1 g/kg) was injected intraperitoneally, and the blood glucose levels at intervals were measured (C). Values are mean ± SD (<i>n</i> = 5). *<i>p</i><0.05, **<i>p</i><0.01, ***<i>p</i><0.001, compared with mock.</p

Gene-expression connection of CYSKT treatments with disease states^*.

<p>*Type 2 diabetic mice were given orally with 200 mg/kg CYSKT for 30 consecutive days. Total RNAs were extracted from muscle tissues and analyzed by microarray.</p>†<p><i>p</i> values were calculated by geneSetTest function implemented in the limma package.</p

Demographic information of participants in this study.

<p>Demographic information of participants in this study.</p

Pathways significantly regulated by CYSKT in type 2 diabetic mice^*.

<p>*Type 2 diabetic mice were given orally with 200 mg/kg CYSKT for 30 consecutive days. Total RNAs were extracted from muscle tissues and analyzed by microarray.</p>†<p>Pathways associated with glucose and lipid metabolism are shown.</p>‡<p><i>p</i> values were calculated by geneSetTest function implemented in the limma package.</p

NF-κB-dependent bioluminescence in living mice and individual organs following 5-FU administration.

<p>(A) <i>In vivo</i> imaging. Transgenic mice were injected intraperitoneally with PBS or 5-FU and imaged at indicated periods. Results are expressed as relative intensity, which is presented as the comparison with the NF-κB-dependent bioluminescent signal relative to mock. Values are mean ± standard error (<i>n</i> = 6 per group). *<i>p</i><0.05, compared with mock. (B) <i>Ex vivo</i> imaging. Transgenic mice were injected intraperitoneally with PBS (mock) or 5-FU. Two days later, mice were sacrificed and organs were subjected to image. The color overlay on the image represents the photon/sec emitted from the organs, as indicated by the color scale. Photos are representative images (<i>n</i> = 6 per group). (C) Quantification of photon emission from the organs. Values are mean ± standard error (<i>n</i> = 6 per group). **<i>p</i><0.01, compared with mock. (D) NF-κB-dependent bioluminescence in the intestine following 5-FU and/or 5-ASA administration. Transgenic mice were administered with 5-FU and/or 5-ASA and imaged 2 days later. The color overlay on the image represents the photon/sec emitted from the intestine, as indicated by the color scale. Photos are representative images (<i>n</i> = 6 per group). Quantification of photon emission from the intestine was shown on the top. Values are mean ± standard error. *<i>p</i><0.05, compared with mock. ^#<i>p</i><0.05, compared with 5-FU treatment.</p