Salusin­α alleviates lipid metabolism disorders via regulation of the downstream lipogenesis genes through the LKB1/AMPK pathway.

Lipid metabolism disorders are a major cause of several chronic metabolic diseases which seriously affect public health. Salusin­α, a vasoactive peptide, has been shown to attenuate lipid metabolism disorders, although its mechanism of action has not been reported. To investigate the effects and potential mechanisms of Salusin­α on lipid metabolism, Salusin­α was overexpressed or knocked down using lentiviral vectors. Hepatocyte steatosis was induced by free fatty acid (FFA) after lentiviral transfection into HepG2 cells. The degree of lipid accumulation was assessed using Oil Red O staining and by measuring several biochemical indices. Subsequently, bioinformatics was used to analyze the signaling pathways that may have been involved in lipid metabolism disorders. Finally, semi­quantitative PCR and western blotting were used to verify the involvement of the liver kinase B1 (LKB1)/AMPK pathway. Compound C, an inhibitor of AMPK, was used to confirm this mechanism's involvement further. The results showed that Salusin­α significantly attenuated lipid accumulation, inflammation and oxidative stress. In addition, Salusin­α increased the levels of LKB1 and AMPK, which inhibited the expression of sterol regulatory element binding protein­1c, fatty acid synthase and acetyl­CoA carboxylase. The addition of Compound C abrogated the Salusin­α­mediated regulation of AMPK on downstream signaling molecules. In summary, overexpression of Salusin­α activated the LKB1/AMPK pathway, which in turn inhibited lipid accumulation in HepG2 cells. This provides insights into the potential mechanism underlying the mechanism by which Salusin­α ameliorates lipid metabolism disorders while identifying a potential therapeutic target.

Overexpression of salusin­ß downregulates adipoR1 expression to prevent fatty acid oxidation in HepG2 cells.

Salusin­ß and adiponectin receptor 1 (adipoR1) serve important roles in the development of certain cardiovascular diseases and lipid metabolism. However, to the best of our knowledge, the relationship between salusin­ß and adipoR1, and their underlying mechanisms of action, currently remain unclear. In the present study, lentiviral vectors designed to overexpress salusin­ß or knock down salusin­ß expression were used in 293T and HepG2 cells. Semi­quantitative PCR was performed to investigate the relationship between salusin­ß and adipoR1 mRNA expression in 293T cells. Western blotting was used to assess the protein expression levels of adipoR1, adenosine monophosphate­activated protein kinase (AMPK), acetyl­CoA carboxylase (ACC) and carnitine palmitoyl transferase 1A (CPT­1A) in transfected HepG2 cells. Simultaneously, HepG2 cells were treated with an adipoR1 inhibitor (thapsigargin) or agonist (AdipoRon) and the resultant changes in the expression levels of the aforementioned proteins were observed. Oil Red O staining and measurements of cellular triglyceride levels were performed to assess the extent of lipid accumulation in HepG2 cells. The results demonstrated that salusin­ß overexpression downregulated adipoR1 expression and inhibited the phosphorylation of AMPK and ACC, which led to decreased CPT­1A protein expression. By contrast, salusin­ß knockdown increased adipoR1 expression and promoted the phosphorylation of AMPK and ACC, which conversely enhanced CPT­1A protein expression. Treatment with adipoR1 agonist, AdipoRon, reversed the effects of salusin­ß overexpression. In addition, salusin­ß overexpression enhanced intracellular lipid accumulation in HepG2 cells induced by free fatty acid treatment. These findings highlighted the potential regulatory role of salusin­ß in adipoR1­mediated signaling pathways. To conclude, the present study provided insights into the regulation of fatty acid metabolism by the liver. In particular, salusin­ß may serve as a potential target for the therapeutic intervention of metabolic disorders of lipids.

Clinical Application of the Classical Theory of Traditional Chinese Medicine in Diabetic Nephropathy.

Objective: To explore the clinical application of the classical theory of traditional Chinese medicine (TCM) in diabetic nephropathy (DN). Methods: A total of 100 patients with DN treated in our hospital from May 2019 to June 2021 were included. The patients were randomly assigned to the control group and the study group. The control group received routine treatment, and the study group was treated with the classical theory of TCM. The efficacy, TCM syndrome score, urine proteinuria (UTP), urine albumin-creatinine ratio (UACR), plasma albumin (ALB), hemoglobin A1c (HbA1C), fasting blood glucose (FBG), blood urea nitrogen (BUN), creatinine (Cr), and treatment safety were compared between the two groups. Results: In comparison to the curative effect, the study group was significantly effective in 34 cases, effective in 12 cases, and ineffective in 4 cases, and the effective rate was 92.00%; the control group was significantly effective in 16 cases, effective in 18 cases, and ineffective in 16 cases, and the effective rate was 68.00%. The effective rate in the study group was higher compared to the control (P < 0.05). In comparison to the TCM syndrome scores, there exhibited no significant difference before treatment (P > 0.05), but after treatment, the TCM syndrome scores of the two groups decreased, and the TCM syndrome scores of the study group were lower compared to the control at 6 weeks, 12 weeks, 24 weeks, and 36 weeks of treatment (P < 0.05). There exhibited no significant difference in the indexes of UTP and UACR before treatment, but the indexes of UTP and UACR in the two groups decreased after treatment, and the indexes of UTP and UACR in the study group were lower compared to the control at 6 and 12 weeks after treatment. There was no significant difference in the indexes of ALB, HbA1C, and FBG before treatment, but after treatment, the indexes of ALB increased, the indexes of HbA1C and FBG decreased in both groups, and the indexes of HbA1C and FBG i4n the study group were lower compared to the control, while the index of ALB in the study group was higher. The indexes of BUN and Cr were compared, and there was no significant difference before treatment, but after treatment, the indexes of BUN and Cr in the two groups decreased, and the indexes of BUN and Cr in the study group were lower compared to the control (P < 0.05). In terms of the treatment safety of the two groups, there was no abnormality in blood, urine, stool routine, and liver and kidney function examination in the study group. No obvious adverse reactions were found in all patients. There were 1 case of abnormal liver function and 2 cases of rash in the control group, and there exhibited no significant difference (P > 0.05). Conclusion: Under the guidance of classical theory, the optimization scheme of comprehensive treatment of TCM may improve renal function by improving metabolic disorders, vascular lesions, neurotrophic disorders, antioxidant stress, and other ways to repair nerve injury, improving the changes of TCM syndromes, signs, and indicators of patients, and delay the progress of DN.