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Objective:To observe the clinical effects of Traditional Chinese Medicine (TCM) Tangnaikang granules, supplementing to conventional treatment methods, on type 2 diabetic foot ulcer.Methods:A total of 67 patients, from Beijing University of Chinese Medicine subsidiary Dongfang Hospital, Tuanjie Lake Community Health Service Center, Qingta Community Health Service Center between August, 2019 and November, 2020 were assigned to treatment group (37 patients) and control group (30 patients) using sealed envelope randomization method. The control group underwent conventional treatment methods that manage blood sugar and treat infections. The treatment group received, in addition to the conventional treatment methods administered to the control group, Tangnaikang granules. Both groups were continuously treated for 90 days. The TCM symptom scores, granulation tissue scores, visual analogue scale (VAS), itching scores and Generic Quality of Life Inventory-74 (GQOL-74) score, wound area were observed pre- and post-treatment. The ELISA kits were used to detect the Vascular Endothelial Growth Factor (VEGF), Epidermal Growth Factor (EGF), Basic Fibroblast Growth Factor (bFGF), fasting blood glucose level, glycated hemoglobin test (HbA1c), fasting insulin level, postprandial 2 hour insulin level, insulin resistance index. And all possible adverse events were noted. The overall clinical treatment effects and TCM symptom treatment effects were assessed.Results:Total effective rate for the treatment group 89.2% (33/37), was significantly different from that of the control group 70.0% (21/30) with χ2=3.90 and P<0.01. The TCM symptom effective rate was 86.5% (32/37) for the treatment group and 63.3% (19/30) for the control group. The difference was statistically different with χ2=4.88 and P<0.01. After treatment, whole blood viscosity, plasma viscosity, fibrinogen and erythrocyte sedimentation rate (ESR) of the treatment group were significantly improved than those of the control group ( t=-23.38, -8.01, 18.10, -18.93,all Ps<0.01). The ulcer closure area (5.43±1.65 cm 2vs. 4.65±1.14 cm 2, t=2.20) and ulcer closure area percentage (59.14%±3.37% vs. 42.42%±3.21%， t=20.63) were statistically different of both groups ( P<0.05 or P<0.01). The VEGF, EGF, and bFGF at 90 day of the treatment group were significantly higher than those of the control group ( t=3.19, 40.59, 28.53, all Ps<0.01). The post-treatment fasting blood glucose level, insulin resistance index of the treatment group were significantly lower than those of the control group ( t=-8.55, -21.38, all Ps<0.01). TCM symptoms of thirsty, feverish sensation in chest, palms and soles in the treatment group were significantly improved than the control group ( χ2=4.38, 4.48, all Ps<0.01). Conclusion:TCM compound Tangnaikang granules can relieve diabetic foot ulcer, reduce wound healing time and improve overall treatment outcome, when it was combined of the conventional treatment.
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Objective: To observe the regulatory effect of Tangnaikang (TNK) on imbalance between neutrophil elastase (NE) and α1-antitrypsin (α1-AT) in ob/ob mice with type 2 diabetes mellitus (T2DM). Method: Thirty-two male SPF ob/ob mice were randomly divided into model group (DM, normal saline) and high-dose TNK group (TNKH, TNK solution 16.04 g·kg-1), middle-dose TNK group (TNKM, TNK solution 8.02 g·kg-1) and low-dose TNK group (TNKL, TNK solution 4.01 g·kg-1). Another 8 C57BL/6J mice were included in normal group (Con, saline). The experiment lasted for four weeks. The general state, body weight (BW) and fasting blood glucose (FBG) of the mice were recorded weekly, the oral glucose tolerance (OGTT) test was performed on the 25th day, the insulin tolerance (ITT) test was performed on the 27th day, and the area under the curve (AUC) was calculated. After the end of the experiment, serum was used to detect the level of fasting insulin (Fins), insulin resistance index (HOMA-IR), total triglyceride (TG), total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL), NE and α1-AT. Adipose tissue was used to detect the expressions of NE, α1-AT, phosphor-insulin receptor substrate 1 antibody (p-IRS1) and glucose transporter 4 (GLUT4) proteins. Result: Compared with the Con group, the BW of the ob/ob mice of the model group increased significantly, the glucose and lipid metabolism indexes showed diabetes, the serum and adipose tissue NE increased significantly (Pα1-AT decreased significantly (PPPogtt and AUCITT were significantly decreased (PPα1-AT increased significantly (PPConclusion: TNK can reduce the BW of ob/ob mice, improve glycolipid metabolism, increase α1-AT level, decrease NE level, and regulate IRS1-GLUT4 signaling pathway, which may be one of its mechanisms in improving IR of adipose tissue mediated by neutrophil.
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The stage of impaired glucose regulation is a necessary stage in the development of diabetes.Nearly a quarter of patients without diabetes intervention develop into diabetes within four to seven years without intervention.Therefore,the intervention of pre-diabetes treatment in the prevention and control of diabetes is of great significance.This study was to investigate the effect of Tangnaikang granule at the early stage of type 2 diabetes mellitus.The age,course of disease,sex,symptom score of TCM were recorded,so were the blood lipid,blood glucose,liver and kidney function,electrocardiogram were recorded.Under the condition of low sugar and low salt diet and healthy exercise,the patients were divided into the control group (n--31) and the treatment group (n=43).The patients in the double-blind control group were treated with Kangnaikang granule and Tangnaikang granule to treat 74 patients with type 2 diabetes mellitus three months.Three months after the detection of fasting and postprandial blood glucose,glycosylated hemoglobin,glucose tolerance,insulin tolerance and other indicators to examine the efficacy of both.Tangnaikang granules were superior to placebo in terms of effective rate of treatment,TCM symptom score and blood sugar recovery rate,and taking Tangnaikang granule without liver and kidney function and other injuries.Tangnaikang is safe and effective for pre-diabetes patients.
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This study aimed to investigate the anti-apoptotic effects of Tangnaikang (TNK) on islet β cells in Zucker diabetic fatty (ZDF) rats.Six male fa/+ ZDF rats were took as the control group,while other thirty male fa/fa ZDF rats were divided into five groups at random:the model group,the metformin group,the high-,mediumand low-dose TNK groups,depending on their body weight and random blood glucose.Prior to the administration,fasting blood glucose and fasting insulin were measured by drawing blood with inner canthusl.Materials were prepared when administered for six weeks.Fasting blood glucose and fasting insulin were detected again.When the sections of the rat pancreatic tissue were embedded,the morphological changes of the islet were observed via HE staining,and the apoptosis of islet β cell were observed using TUNEL.Positive expression of Caspase-3,the transduction enzyme of cell death signal,was tested by immunohistochemical method.It was found that the fasting blood glucose of the (fa/fa) ZDF rats in the high-,medium-and low-dose TNK group was significantly improved after administration (P ＜ 0.01).The serum insulin of rats in the high-,medium-and low-dose TNK group arised compared with the model group,while the high-and low-dose TNK group showed differences in a statistical sense.Compared with the model group,the HOMA-IR of all the treatment group decreased,while significant difference was presented between the high-dose TNK group and the metformin group.HE staining showed that the morphology of the islet β cell of the rats in all the treatment group was improved.The results of TUNEL showed significantly apoptotic changes on islet β cell of the fa/fa ZDF rats.Compared with the model group,the positive expression of TUNEL in the metformin group and the high-dose TNK group were significantly reduced (P ＜ 0.05).The result of immunohistochemistry method showed that the protein levels of Caspase-3 in the metformin group and the high-dose TNK group decreased (P ＜ 0.05).In conclusion,it was demonstrated that TNK effectively reduced the apoptosis of islet β cells in fa/fa ZDF rats,which presented a protective effect.
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This article was aimed to study the content determination methods ofTang-Nai-Kang (TNK) granules. The content of rosemary acid in TNK granules was determined by HPLC method, using C18 chromatographic column (150 mm×4.6 mm, 5μm), the mobile phase of 0.1% trifluoroacetic acid - methanol (60:40),λ of 330 nm. The theoretical plate number was more than 2 000. The standard curve method was used in the determination. Contents of ginsenosides Rg1, Re and Rb1 were determined by HPLC method, using C18 chromatographic column (150 mm× 4.6 mm, 5μm), the mobile phase of water-acetonitrile, gradient elution method,λ of 203 nm. The theoretical plate number was more than 2 000. The standard curve method was used in the determination. The results showed that rosmarinic acid was linear in the range of 0.300-0.500 mg·mL-1 with good linearity. The regression equation wasY = 1E + 07X -7 334,R2 = 0.999. Its sample recovery was 97.32% (RSD 1.25%). Ginsenoside Rg1 was linear in the range of 0.138-0.220 mg·mL-1 with good linearity. The regression equation wasY = 2E + 06X +34 864,R2 = 0. 999. Ginsenoside Re was linear in the range of 0.126-0.250 mg·mL-1 with good linearity. The regression equation wasY = 2E + 06X + 39 879,R2 = 0. 999. Ginsenoside Rb1 was linear in the range of 0.132-0.220 mg·mL-1 with good linearity. The regression equation wasY = 2E + 06X + 39 070,R2 =0. 999. Their sample recoveries were Rg1 97.97% (RSD 1.83%), Re 101.80% (RSD 1.83%), Rb1 98.35% (RSD 1.82%). It was concluded that the content determination method established for the quality control of TNK granules was simple, fast and reliable.
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This study was aimed to explore the effect of Tang-Nai-Kang (TNK) on trans-differentiation of renal tubular epithelial cell in KKAy mice in order to discuss the possible mechanism. Fifty 12-week-old male KKAy mice were randomly divided into the model group, valsartan group, TNK high-dose, middle-dose and low-dose group, with 10 rats in each group. Ten C57BL/6J mice were used in the normal group. Rats in the model group and normal group were given 0.9% sodium chloride solution. Rats in other groups were given the corresponding drugs. After 8 weeks of gavage administration, kidneys of all mice were sampled and given Mosson and PAS dyeing. Expression distribution of α-smooth muscle actin (α-SMA) and E-cadherin in kidney tissues were observed under immunohistochemical staining. Expression of transforming growth factor-β1 (TGF-β1) was measured by western blot. The results showed that compared with the normal group, the area of renal fibrosis in the model group was significantly increased (P < 0.01); the expression of α-SMA was stronger; and the expression of E-cadherin was weaker. Compared with the model group, the area of renal fibrosis in the valsartan group, TNK high-dose, middle-dose and low-dose groups were significantly decreased (P< 0.01); the expression of α-SMA was weaker (P< 0.01);and the expression of E-cadherin was obviously increased (P < 0.05). The TGF-β1 expression in the model group was significantly higher than that in the normal group (P < 0.01). Compared with the model group, the TGF-β1 expression in the valsartan group, TNK low-dose, middle-dose and high-dose groups were significantly lowered (P<0.01). And the TGF-β1 expression in the TNK high-dose group was even lower than that in the valsartan group. It was concluded that TNK was able to suppress the epithelial-mesenchymal transition (EMT) of renal tubular epithelial cell, and lessen the renal tubule interstitial fibrosis, in order to protect the kidney.
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Objective To explore the effect of Tangnaikang (TNK) on extracellular matrix expression of human tubular epithelial cell HK-2 induced by TGF-β1 and explore its mechanism on the renal fibrosis. Methods The HK-2 cells were cultured by DMEM/F12 (1∶1) with 10%fetal bovine serum and divided into control group, TGF-β1 group (TGF-β110 ng/mL), rat serum control group (TGF-β110 ng/mL +10% rat serum), TNK-containing rat serum therapy groups (TGF-β110 ng/mL+5% Tangnaikang, or +10%Tangnaikang, or +20% TNK). After 24 h of administration, the expression of ColⅠ, Col Ⅲ and FN mRNA were tested by fluorescence quantitative PCR assay. Results The expression of ColⅠ, Col Ⅲ and FN mRNA of HK-2 cultured with TGF-β1 were much higher than the control, and significantly decreased in HK-2 cultured with TGF-β1 plus Tangnaikang compared with only TGF-β1 (P<0.05), but rat serum control had no effect. Conclusion TNK could inhibit the expression of ColⅠ, Col Ⅲ and FN mRNA of HK-2 cell induced by TGF-β1, and prevent the development of renal fibrosis to some extent.
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Objective Through studying the effect of Tangnaikang (TNK) on the expression of Smad 2, 3, 7 mRNA of human renal tubular epithelial cells HK-2 induced by transforming growth factor-β1 (TGF-β1), to explore the mechanism of TNK on prevention and treatment of renal fibrosis. Methods The HK-2 cells were cultured by DMEM/F12 (1∶1) with 10% fetal bovine serum and divided into control group, TGF-β1 group (TGF-β1 10 ng/mL), blank serum control group (TGF-β1 10 ng/mL + 10% animal serum), TNK drug-containing serum therapy groups (TGF-β1 10 ng/mL + 5% TNK or + 10% TNK or + 20% TNK). After 24 h, the expression of Smad 2, 3, 7 mRNA were tested by fluorescence quantitatiye PCR assay. Results After the HK-2 cells were induced by TGF-β1, the expression of Smad 2, 3 mRNA were increased and the expression of Smad 7 mRNA was decreased compared with the control group (P<0.05). The expression of Smad 2, 3 mRNA were decreased and the expression of Smad 7 mRNA was increased in TNK drug-containing serum therapy groups compared with TGF-β1 group (P<0.05), but blank serum control group had no such effect. Conclution TNK could prevent the development of renal fibrosis to some extent through regulating the expression of Smads signaling pathway.
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Objective To investigated the effect of TangNaiKang (TNK) on VEGF protein expression of GK rats Thoracic aorta. Methods 51 male GK rats were divided into five groups randomly: model group, pioglitazone group, and TNK treatment group (low, immediate and high dose). Another 10 male Wistar rats were served as normal control group. GK rats were fed with high-grease forage, while normal control group was fed with a standard diet. Fasting blood glucose, general HE staining and VEGF protein expression were detected by immunohistochemistry. Results The fasting glucose had a significant decline in TNK treatment groups. HE staining showing TNK can ameliorate intima thickness, reduce hyperplasia of shallow vascular smooth muscle cell, and improve wavy and plexiform arrangement of elastic lamina. Immunohistochemistry also showed that TNK decreased VEGF protein expression of great vessels. Conclusion TangNaiKang can prevent and cure diabetic vascular complication of GK rats.
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Objective To observe the effects of Tangnaikang on interleukin 6 (IL-6) and minor necrosis factor-α(TNF-α) levels of Diabetic KKAy mice. Methods After diabetic KKAy mice were treated with Tangnaikang for 8 weeks,FBG was determined by glucose test-paper, IL-6 and TNF-αconcentratious in serum and adipocytes were determined by ELISA. Results IL-6 and TNF-α levels were reduced in serum and adipocytes of KKAy mice. Conclusion Tangnaikang can significantly reduce the levels of IL-6 and TNF-α in Diabetic KKAy mice, and the reduction of them can help to decrease FBG level.