Integrated metabolomics and transcriptomics analysis reveals the mechanism of Tangbi capsule for diabetic lower extremities arterial disease.

Objective: Tangbi capsule (TBC) is a traditional Chinese medicine prescription, which has the potential to improve the vascular insufficiency of lower extremities and limb numbness in diabetes. However, the potential mechanism remains unknown. This study aims to investigate the pharmacological effects and mechanism of TBC on rats with diabetic lower extremities arterial disease (LEAD). Methods: The mechanism of TBC on diabetic LEAD was investigated through metabolomics and transcriptomics analysis, and the main components of TBC were determined by mass spectrometry. The efficacy and mechanism of TBC on diabetic LEAD rats were investigated through in vitro experiments, histopathology, blood flow monitoring, western blot, and real-time polymerase chain reaction. Results: Mass spectrometry analysis identified 31 active chemical components in TBC including (2R)-2,3-Dihydroxypropanoic acid, catechin, citric acid, miquelianin, carminic acid, salicylic acid, formononetin, etc. In vitro analysis showed that TBC could reduce endothelial cell apoptosis and promote angiogenesis. Histopathological analysis showed that TBC led to an obvious improvement in diabetic LEAD as it improved fibrous tissue proliferation and reduced arterial wall thickening. In addition, TBC could significantly increase the expression levels of HIF-1α, eNOS, and VEGFA proteins and genes while reducing that of calpain-1 and TGF-ß, suggesting that TBC can repair vascular injury. Compared with the model group, there were 47 differentially expressed genes in the whole blood of TBC groups, with 25 genes upregulated and 22 downregulated. Eighty-seven altered metabolites were identified from the serum samples. Combining the changes in differentially expressed genes and metabolites, we found that TBC could regulate arginine biosynthesis, phenylalanine metabolism, pyrimidine metabolism, arachidonic acid metabolism, pyrimidine metabolism, arachidonic acid metabolism, nucleotide metabolism, vitamin B6 metabolism and other metabolic pathways related to angiogenesis, immune-inflammatory response, and cell growth to improve diabetic LEAD. Conclusion: TBC improved vascular endothelial injury, apoptosis, lipid accumulation, liver and kidney function, and restored blood flow in the lower extremities of diabetic LEAD rats. The mechanism of TBC in the treatment of diabetic LEAD may be related to the modulation of inflammatory immunity, lipid metabolism, and amino acid metabolism. This study presented preliminary evidence to guide the use of TBC as a therapy option for diabetic LEAD.

Effect and Safety of Herbal Medicine Foot Baths in Patients with Diabetic Peripheral Neuropathy: A Multicenter Double-Blind Randomized Controlled Trial.

OBJECTIVE: To evaluate the effect and safety of foot baths with Tangbi Waixi Decoction (TW) in treating patients with diabetic peripheral neuropathy (DPN). METHODS: It is a multicenter double-blinded randomized controlled trial. Participants with DPN were recruited between November 18, 2016 and May 30, 2018 from 8 hospitals in China. All patients received basic treatments for glycemic management. Patients received foot baths with TW herbal granules either 66.9 g (intervention group) or 6.69 g (control group) for 30 min once a day for 2 weeks and followed by a 2-week rest, as a therapeutic course. If the Toronto Clinical Scoring System total score (TCSS-TS) ⩾6 points, the patients received a total of 3 therapeutic courses (for 12 weeks) and were followed up for 12 weeks. The primary outcome was change in TCSS-TS score at 12 and 24 weeks. Secondary outcomes included changes in bilateral motor nerve conduction velocity (MNCV) and sensory nerve conduction velocity (SNCV) of the median and common peroneal nerve. Safety was also assessed. RESULTS: Totally 632 patients were enrolled, and 317 and 315 were randomized to the intervention and control groups, respectively. After the 12-week intervention, patients in both groups showed significant declines in TCSSTS scores, and significant increases in MNCV and SNCV of the median and common peroneal nerves compared with pre-treatment (P<0.05). The reduction of TCSS-TS score at 12 weeks and the increase of SNCV of median nerve at 24 weeks in the control group were greater than those in the intervention group (P<0.05). The number of adverse events did not differ significantly between groups (P>0.05), and no serious adverse event was related with treatment. CONCLUSION: Treatment of TW foot baths was safe and significantly benefitted patients with DPN. A low dose of TW appeared to be more effective than a high dose. (Registry No. ChiCTR-IOR-16009331).

[Comparative study of enhancing effect on mRNA expression of hematopoietic growth factors in rat bone marrow mesenchymal stem cells by ginseng polysaccharide and ginsenoside].

OBJECTIVE: To study mRNA expression levels of main hematopoietic growth factors in bone marrow mesenchymal stem cells (BM-MSC), and to compare effect on mRNA expression levels treated by ginseng polysaccharide and ginsenoside. METHODS: Relative quantification real-time polymerase chain reaction (RT-PCR) was used to observe mRNA expression levels of IL4, Csf2, Kitlg, Csf1, IL6, Lif, Csf3, IL11, Epo, and IL3, etc. in rat BM-MSC treated with ginseng polysaccharide (20 microg/mL) or ginsenoside (20 microg/mL) at 12, 24, and 36 h. RESULTS: IL4 and Csf2 mRNA expressions were not detected. Relative expression of Kitlg, Csf1, IL6, Lif, Csf3, IL11, Epo and IL3 mRNA ranked in an attenuating order when compared with Gapdh mRNA. mRNA expression of Epo and IL3 was not significantly changed at any time point by treatment of ginseng polysaccharide or ginsenoside in rat BM-MSC (P > 0.05). mRNA expression of Csf1, IL6, Lif, Csf3 and IL11 were significantly enhanced at 12 and 36 h by treatment of ginseng polysaccharide (P < 0.05) and that of Csf1, IL6, Lif, Csf3, and Kitlg were significantly enhanced at 24 h in rat BM-MSC (P < 0.05). The enhanced mRNA expression was Csf3 at 12 h, Csf3, IL6 and Lif at 24 h, and Csf3, IL6, Lif, IL11, and Kitlg, respectively at 36 h by treatment of ginsenoside in rat BM-MSC. CONCLUSIONS: The enhancement of ginseng polysaccharide was stronger than that of ginsenoside on mRNA expression of hematopoietic growth factors in the initial stage. As time went by, the enhancement of ginsenoside gradually increased and exceeded that of ginseng polysaccharide.