Regulation of Bcl-2 and the NF-kB Signaling Pathway by Succinyl Rotundic Acid in Livers of Rats with Alcoholic Hepatitis

In this study, the protective effects of succinyl rotundic acids on alcoholic hepatitis in irradiated rats as well as the effects of Bcl-2-Bax-caspase-3 and the NF-kB signal pathways were studied. SD rats were divided into four groups randomly: normal; model; and succinyl rotundic acid low-, middle-, and high-dose groups. Distilled water, 60% ethanol and 60% ethanol +SRA, respectively, were given for 30 days. ELISA was used to measure serum levels of LDH, AST, ALT, NOS, NO, MDA, GSH and TG. Western blotting was used to measure protein levels of Bcl-2, Bax, caspase-3, NF-kB p 65, IKBA, HO-1, Nrf2 and CYP2E1. Compared with the model group, LDH, AST, ALT, NOS, NO, MDA and TG levels were lower in serum of low-, middle-, and high-dose groups (P < 0.01, P < 0.05 and P < 0.05 in all); GSH content was greater in serum of low-, middle- and high-dose groups (P < 0.05). Levels of Bcl-2, HO-1, and Nrf2 were greater (P < 0.01 in all); those of Bax, caspase-3, NF-kB p65, IKBA, and CYP2E1 were lower (P < 0.01 and P < 0.001 in all). These findings suggest that succinyl rotundic acid reduces inflammatory reactions by reducing levels of NOS and NO, regulating levels of Bcl-2, Bax, caspase-3, NF-kB, and anti-oxidative stress pathways, and has an antagonistic effect on alcoholic liver injury. The agent has potential to treat clinical alcoholic liver disease. © 2021 Friends Science Publishers

Rotundic Acid Regulates the Effects of Let-7f-5p on Caco2 Cell Proliferation

Background & Objective: Nowadays, the interaction between natural products and microRNAs provides a promising field for exploring the chemo preventive agents for various cancers.As a member of microRNAs, the expression of let-7f-5p is universally down regulated in colorectal cancer (CRC). The present study aimed to uncover the function of let-7f-5p in the proliferation of human colon cancer cell line Caco2 and explored chemo preventive agents from natural resources that can prevent the development of CRC. Methods: Herein, Caco2 cells were transfected with let-7f-5p mimic and inhibitor to manipulate let-7f-5p levels, and the expression of let-7f-5p wasper formed by RT‑qPCR. Next, we determined how let-7f-5p regulates Caco2 cell proliferation by using MTT, wound-healing, cell cycle,and colony formation assays.Besides, to further understand the effect of let-7f-5p, we evaluated the protein level of AMER3 and SLC9A9 by using western blotting assays. Results: The results showed a suppressive function of let-7f-5p on Caco2 cell proliferation and then put forward a triterpenoid (rotundic acid, RA) which significant antagonized the effect of cell proliferation, restitution after wounding,and colony formation caused by let-7f-5p. Moreover, the western blot results further indicated that the inhibitory effect of RA might be due to its suppressive role in let-7f-5p-targeted AMER3 and SLC9A9 regulation. Conclusion: Our validation study results confirmed that let-7f-5p was a potent tumor suppressor gene of Caco2 cell proliferation,and RA showed as a regulator of the effect oflet-7f-5p on cell proliferation and then could be a potential chemo preventive agent for CRC treatment.

Succinyl rotundic acid inhibits growth and promotes apoptosis in the HeLa cervical cancer cell line

Rotundic acid (RA) is a kind of pentacyclic triterpene saponins, which widely exists in holly and other plants. It has anti-tumor and lipid-lowering effects, and has been widely used in the prevention and treatment of vascular diseases. Succinyl RA (SRA) is a kind of circular acid derivative with strong water solubility. However, SRA’s effect and mechanism about the therapy of cervical cancer is still not clarified. The inhibitory effect of SRA on Hela cells and its molecular mechanism were investigated in this study. Hela cell line was used to establish an in vitro model to study the effect of SRA. The effects of SRA on cell proliferation, cell cycle and apoptosis were analyzed by flow cytometry and detected by 3-(4,5-dimethyl thiazole-2-yl)-2, 5-diphenyltetrazole bromide. The experimental results show that SRA can inhibit the growth of Hela and diffusion, inducing cell apoptosis. SRA can also reduce the expression of CyclinD 1 and p21. It was also decreased the expression of apoptosis inhibitor B cell lymphoma (Bcl-2), and increased the expression of Bcl-2-related X-protein and Caspase-3. This study provides evidence for the progress of SRA in inhibiting Hela cells and presents potential applications of SRA as an alternative therapy for cervical cancer.

Rotundic Acid Protects against Metabolic Disturbance and Improves Gut Microbiota in Type 2 Diabetes Rats

Rotundic acid (RA) is a major triterpene constituent in the barks of Ilex rotunda Thunb, which have been widely used to make herbal tea for health care in southern China. RA has a variety of bioactivities such as anti-inflammation and lipid-lowering effect. However, little is known about the effects and mechanisms of RA on metabolic disturbance in type 2 diabetes (T2D) and its effect on gut microbiota. A T2D rat model induced by high fat diet (HFD) feeding and low-dose streptozotocin (STZ) injection was employed and RA showed multipronged effects on T2D and its complications, including improving glucolipid metabolism, lowering blood pressure, protecting against cardiovascular and hepatorenal injuries, and alleviating oxidative stress and inflammation. Furthermore, 16s rRNA gene sequencing was carried out on an Illumina HiSeq 2500 platform and RA treatment could restore the gut microbial dysbiosis in T2D rats to a certain extent. RA treatment significantly enhanced the richness and diversity of gut microbiota. At the genus level, beneficial or commensal bacteria Prevotella, Ruminococcus, Leuconostoc and Streptococcus were significantly increased by RA treatment, while RA-treated rats had a lower abundance of opportunistic pathogen Klebsiella and Proteus. Spearman’s correlation analysis showed that the abundances of these bacteria were strongly correlated with various biochemical parameters, suggesting that the improvement of gut microbiota might help to prevent or attenuate T2D and its complication. In conclusion, our findings support RA as a nutraceutical agent or plant foods rich in this compound might be helpful for the alleviation of T2D and its complications through improving gut microbiota.