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Objective:The purpose of this study was to explore the therapeutic effect of modified Xiaoke prescription on patients with Yin deficiency and heat excessive type 2 diabetes mellitus (T2DM), and its influence on TCM syndrome scores, pancreatic islet function and oxidative stress.Methods:Randomized controlled trial. Eighty patients with Yin deficiency and heat excessive T2DM treated in the hospital between January and July 2021 were selected, and divided into observation group (41 cases) and control group (39 cases) by random number table method. Patients in the control group were treated with conventional western medicine, and patients in the observation group were treated with modified Xiaoke Prescription on the basis of the control group. Both groups were treated for 1 month. TCM syndrome scores were performed before and after treatment. Fasting plasma glucose (FPG) and 2 hPG were measured by glucose oxidase method. Serum HbA1c, malondialdehyde (MDA), 8-hydroxy-2'-deoxyguanosine (8-OHdG) levels and SOD activity were measured by ELISA. The levels of low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C) and total cholesterol (TC) were detected by colorimetry.Results:The total effective rate of the observation group was 92.68% (38/41), and that of the control group was 76.92% (30/39). The difference between the two groups was statistically significant ( χ2=3.89, P=0.048). After treatment, the scores of tiredness and fatigue, thirst and appetite, overeating and hunger, redness of tongue and lack of saliva and total scores in the observation group were significantly lower than those in the control group ( t=4.46, 16.89, 13.37, 8.58, 8.38, P<0.01). After treatment, the levels of serum FPG [(7.31±0.90) mmol/L vs. (8.72±1.50) mmol/L, t=5.13], 2 hPG [(9.64±2.05) mmol/L vs. (12.85±1.20) mmol/L, t=8.49], HbA1c [(7.64±0.58)% vs. (8.11±1.35)%, t=2.04] in the observation group were significantly lower than those in the control group ( P<0.05); MDA [(3.96±1.00) mmol/L vs. (5.04±0.73) mmol/L, t=5.49], 8-OHdG [(203.41±30.70) ng/L vs. (234.50±59.00) ng/L, t=2.98] levels were significantly lower than those in the control group ( P<0.05); The activity of serum SOD [(48.64±5.05) mU/L vs. (41.75±3.58) mU/L, t=7.01] was significantly higher than that of the control group ( P<0.01); The serum LDL-C [(2.01±0.11) mmol/L vs. (2.56±0.25) mmol/L, t=12.84], TC [(4.75±0.20) mmol/L vs. (5.12±0.07) mmol/L, t=10.93] levels were significantly lower than those in the control group ( P<0.01); The serum HDL-C [(1.62±0.18) mmol/L vs. (1.24±0.42) mmol/L, t=5.31] level was significantly higher than that of the control group ( P<0.01). Conclusion:The modified Xiaoke Prescription can improve clinical symptoms, curative effect and pancreatic function, and relieve oxidative stress on the patients with T2DM.
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ObjectiveTo investigate the effect of Zuoguiwan on pancreatic islet function in offspring of gestational diabetes mellitus （GDM） maternal rat model and explore the mechanisms of Zuoguiwan in improving pancreatic islet function based on postpartum pancreatic regeneration. MethodHealthy female SD rats with normal blood glucose levels were paired with male rats in a 2∶1 ratio and housed together. Pregnancy was confirmed based on vaginal plugs or vaginal smears. The pregnant rats were divided into the following groups： normal group， model group， insulin group （insulin Detemir， 20 U·kg-1）， low-dose Zuoguiwan group （1.89 g·kg-1）， and high-dose Zuoguiwan group （3.78 g·kg-1）. The GDM rat model was induced using streptozotocin in rats except for those in the normal group. The model was confirmed by blood glucose testing in the maternal rats. Except for the normal and model groups， the other groups received daily administration of corresponding treatments. At 21 days after birth， fasting blood glucose （FBG） and fasting serum insulin （FINS） levels were measured in 6 offspring from each group. The homeostasis model assessment of insulin resistance （HOMA-IR） was calculated， and an oral glucose tolerance test （OGTT） was performed on additional 12 offspring from each group. Blood samples were taken from the abdominal aorta of the offspring at postnatal day 22， and enzyme-linked immunosorbent assay （ELISA） was used to measure insulin， glucagon （GC）， pancreatic polypeptide （PPY）， and somatostatin （SS） levels in the serum. Hematoxylin-eosin （HE） staining was performed to observe pathological changes in the pancreatic tissue of the offspring. Immunofluorescence （IF） was used to observe the area and structure of the pancreatic islets. Western blot was used to detect the expression of key proteins involved in the development and functional expression of pancreatic β-cells， namely pancreatic and duodenal homeobox factor 1 （Pdx1）， Nkx6.1， and Glucose transporter 2 （Glut2）. ResultCompared with the normal group， the model group showed significant increases in FBG and FINS levels， and HOMA-IR （P<0.01）. Compared with the model group， the insulin group showed significant decreases in FBG levels and HOMA-IR （P<0.01）， the low-dose Zuoguiwan group showed a significant decrease in FBG levels （P<0.05）， and the high-dose Zuoguiwan group showed significant decreases in FBG and FINS levels， and HOMA-IR （P<0.01）. Compared with the normal group， the model group showed significant increases in OGTT 60-min blood glucose levels and AUC index （P<0.05， P<0.01）. Compared with the model group， the high-dose Zuoguiwan group showed significant decreases in OGTT60-min blood glucose levels and area under the curve（AUC） index （P<0.05， P<0.01）. HE staining of pancreatic tissue showed that compared with the normal group， the model group had a reduced number of islets and a loose arrangement of acinar cells. Compared with the model group， the groups with drug treatment showed increased number of islets and a compact arrangement of acinar cells. Compared with the normal group， the model group had significantly increased levels of insulin， GC， PPY， and SS in the serum （P<0.01）. Compared with the model group， the low-dose and high-dose Zuoguiwan groups and the insulin group showed significantly decreased serum levels of insulin， GC， PPY， and SS （P<0.05， P<0.01）. IF results showed that compared with the normal group， the model group had a significantly lower positive rate of insulin （P<0.05）. Compared with the model group， the low-dose and high-dose Zuoguiwan groups showed a significant increase in the positive rate of insulin （P<0.05）. There was no significant difference in the positive rate of GC among the groups. In terms of the proportion of insulin and GC in individual islets， compared with the normal group， the model group showed a significant decrease in the proportion of insulin （P<0.01） and a significant increase in the proportion of GC （P<0.01）. Compared with the model group， the low-dose and high-dose Zuoguiwan groups showed significantly increased proportion of insulin （P<0.01） and significantly decreased proportion of GC （P<0.01）. Compared with the normal group， the model group showed significantly decreased expression levels of Pdx1， Nkx6.1， and Glut2 proteins in the pancreatic tissue of GDM offspring （P<0.05）. Compared with the model group， the insulin group and the low-dose Zuoguiwan group showed significant increases in the expression levels of Pdx1 and Nkx6.1 proteins in the pancreatic tissue of GDM offspring （P<0.05）， and the low-dose and high-dose Zuoguiwan groups showed significant increases in the expression levels of Glut2 protein （P<0.05）. ConclusionZuoguiwan can promote pancreatic islet development in offspring of GDM maternal rat model， improve pancreatic islet morphology and function， and alleviate insulin resistance. Its mechanism of action may be related to the regulation of Pdx1， Nkx6.1， and Glut2 protein expression in the pancreatic tissue of offspring.
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Objective To investigate the isolation and protective effect of a new islet purification solution (IPS)-Optiprep solution on the islet in mouse models. Methods The digested pancreatic islets were divided into the IPS and UW groups according to the islet volume. The pancreatic islets were isolated by the continuous gradient density centrifugation using IPS-Optiprep or UW-Optiprep solutions. The purification efficiency and isolated islet activity of purification solution were compared between two groups. The diabetic mouse models were successfully induced and randomly assigned into three groups. In the experimental group (n=10), the mice received pancreatic islet transplantation using islets isolated and purified by the IPS-Optiprep solution. In the control group (n=10), the mice underwent pancreatic islet transplantation using islets isolated and purified by the UW-Optiprep solution. In the sham surgery group (n=5), the mice merely underwent surgery without pancreatic islet transplantation. Postoperative blood glucose levels were detected and compared among three groups. The blood glucose levels of intraperitoneal glucose tolerance test at postoperative 21 d were statistically compared between the experimental and control groups. The cost of the preparation of two isolation solutions was also compared. Results Compared with the UW group, the islet equivalent (IEQ), islet purity, recovery rate and islet integrity were significantly higher in the IPS group. Islet morphological observation revealed that the islet membrane was complete and the islet diameter in the IPS group was considerably larger than that in the UW group. The activity of purified islets in the UW group was significantly higher than that in the IPS group [(88±5)% vs. (84±3)%, P<0.01]. Compared with the UW-Optiprep solution, identical in vivo islet function was obtained in the IPS-Optiprep solution.The cost of IPS-Optiprep solution was significantly less than that of the UW-Optiprep solution. Conclusions The new IPS-Optiprep solution yields higher islet isolation efficiency, purification, integrity and recovery rate and significantly reduces the purification cost compared with the UW-Optiprep solution. Nevertheless, IPS-Optiprep solution exerts a less protective effect on the activity of islet cells, which is probably correlated with the high islet integrity and the endotoxin in the IPS-Optiprep solution.
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This study was aimed to evaluate effect of Qinghua Granules (QHG) on glycometabolism, pancreatic islet function and oxidative stress in type-2 diabetics with heat syndrome. A total of 60 cases of type-2 diabetics with heat syndrome (according to the Syndrome Element Syndrome Differentiation) were enrolled in the clinic of the Department of Endocrinology and Metabolism, Shuguang Hospital Affiliated to Shanghai University of Tradi-tional Chinese Medicine. The average age of enrolled cases was (57.9 ± 6.9) years. Enrolled cases were randomly divided into the treatment group and the control group. The original hypoglycemic plan was continued to use. In the treatment group, QHG was administrated. And in the control group, placebo was given. The administration dosage in both groups was one package per day. The treatment course was 12 weeks. The fasting and postpran-dial (120 min after standard meal) blood samples before and after medication were collected. The main evalua-tion indexes were fasting plasma glucose (FPG), postprandial plasma glucose (PPG) and hemoglobin A1c (HbA1c). The secondary evaluation indexes were homeostasis model assessment (HOMA2-%B, HOMA2-%S, HOMA2%-IR), superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), maleic dialdehyde (MDA), total cholesterol (TC), triglyceride (TG), high-density lipoprotein (HDL) and low-density lipoprotein (LDL). The anal-ysis of variance was used in the comparison of efficacy between two groups . The results showed that HbA1c in the treatment group was obviously reduced, and HOMA2-%B was obviously increased. There was no significant changes in the control group ( P = 0 . 044 , P = 0 . 016 ) . In the treatment group , SOD increased obviously , MDA reduced obviously. There was no significant change in the control group. There was difference b etween two groups (P = 0.011, P = 0.049). There was no change on blood lipids or other evaluation indexes. It was conclud-ed that QHG is effective in the improvement of glycometabolism, islet β-cell functions and oxidative stress in type-2 diabetics with heat syndrome .