Decoding the key compounds and mechanism of Shashen Maidong decoction in the treatment of lung cancer.

BACKGROUND: Lung cancer is a malignant tumour with the fastest increase in morbidity and mortality around the world. The clinical treatments available have significant side effects, thus it is desirable to identify alternative modalities to treat lung cancer. Shashen Maidong decoction (SMD) is a commonly used traditional Chinese medicine (TCM) formula for treating lung cancer in the clinic. While the key functional components (KFC) and the underlying mechanisms of SMD treating lung cancer are still unclear. METHODS: We propose a new integrated pharmacology model, which combines a novel node-importance calculation method and the contribution decision rate (CDR) model, to identify the KFC of SMD and to deduce their mechanisms in the treatment of lung cancer. RESULTS: The enriched effective Gene Ontology (GO) terms selected from our proposed node importance detection method could cover 97.66% of enriched GO terms of reference targets. After calculating CDR of active components in key functional network, the first 82 components covered 90.25% of the network information, which were defined as KFC. 82 KFC were subjected to functional analysis and experimental validation. 5-40 µM protocatechuic acid, 100-400 µM paeonol or caffeic acid exerted significant inhibitory activity on the proliferation of A549 cells. The results show that KFC play an important therapeutic role in the treatment of lung cancer by targeting Ras, AKT, IKK, Raf1, MEK, and NF-κB in the PI3K-Akt, MAPK, SCLC, and NSCLC signaling pathways active in lung cancer. CONCLUSIONS: This study provides a methodological reference for the optimization and secondary development of TCM formulas. The strategy proposed in this study can be used to identify key compounds in the complex network and provides an operable test range for subsequent experimental verification, which greatly reduces the experimental workload.

Protective Effects and Mechanisms of Shenhua Tablet () on Toll-Like Receptors in Rat Model of Renal Ischemia-Reperfusion Injury.

OBJECTIVES: To investigate the protective effects and potential mechanisms of Shenhua Tablet (, SHT) on the toll-like receptors (TLRs)-mediated signaling pathways in a rat model of kidney ischemia-reperfusion injury (IRI). METHODS: Sixty male Wistar rats were randomly divided into 5 groups: sham surgery, model control, astragaloside (150 mg•kg-1•d-1), low- and high-dose SHT (1.5 and 3.0 g•kg-1•d-1, repectively) groups. One week after drug treatment, rats underwent surgery to establish the IRI models. At 24 h and 72 h after the modeling, serum creatinine (Scr) and blood urea nitrogen (BUN) were analyzed; pathological damage were scored after periodic acid-Schiffstaining. TLR2, TLR4 and myeloid differentiation factor 88 (MyD88) protein and mRNA expressions were detected by inmmunohistochemistry, Western blot and qPCR. Tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6) protein expressions were detected by enzyme linked immunosorbent assay. RESULTS: Compared with the sham group, the model group exhibited severe change in renal function (Scr: 189.42±21.50, P<0.05), pathological damage (damage score: 4.50±0.55, P<0.05), and the expression levels of TLR2, TLR4, MyD88, TNF-α, IL-6 were significantly higher than other groups. Meanwhile, the levels of TLRs in model group showed upward tendency from 24 to 72 h, unparalleled with pathological and functional changes. The aforementioned parameters were alleviated to a certain extent, and, in addition to TLRs, presented the obvious downward trending from the 24 to 72 h after the intervention in the SHT and astragaloside groups relative to the model (P<0.05); in particular, the most significant mitigation of these changes was observed in the SHT-H group (P<0.05). CONCLUSION: TLRs may be an important spot to treat and research in acute kidney injury. SHT could effectively mitigate renal injuries and promote recovery of IRI injuries through suppression of degeneration induced by up-regulation of TLR2 and TLR4 expression levels in the MyD88-dependent signaling pathway and exhibit some dose dependence.

Protective effects of astragaloside in rats with adriamycin nephropathy and underlying mechanism.

The present study was designed to determine the mechanism underlying the treatment of nephrotic syndrome using astragaloside by observing the effects of astragaloside on the expression of nephrin and podocin proteins and genes in kidneys of rats with adriamycin nephropathy. The rats were injected with adriamycin and, after successful model establishment, randomly divided into a model group, a Methylprednisolone (MP) group, and an astragaloside group. The 24-h complete urine samples were collected. Biochemical indicators were monitored, and kidney tissues were collected for pathological analysis using light microscopy and electron microscopy. The mRNA expression of nephrin and podocin was measured in the kidney tissues using the real-time qPCR, and the protein expression levels of nephrin and podocin were detected using Western blot analysis. At the end of 12 weeks of drug intervention, the urinary protein level was lower in the MP and astragaloside groups than that in the model group (P = 0.008 and P = 0.01, respectively). Serum albumin was higher in the MP and astragaloside groups than in the model group (P < 0.001 and P = 0.012, respectively). Podocytes in the MP group were nearly normal, and fusion of podocytes in the astragaloside group was significantly less than that in the control group. The nephrin and podocin mRNA and protein expression levels in the intervention groups were higher (P < 0.05) than that in the model group. Due to the increased expression of podocyte-related nephrin and podocin proteins, astragaloside maintained slit diaphragm integrity and decreased the level of proteinuria in rats with adriamycin nephropathy.

Upregulation of arginase activity contributes to intracellular ROS production induced by high glucose in H9c2 cells.

Arginase is upregulated in some tissues under diabetes states. Arginase can compete with nitroxide synthase (NOS) for the common substrate L-arginine and thus increases oxidative stress by NOS uncoupling. We want to analyze whether arginase is upregulated and contribute to oxidative stress in H9c2 cells during high glucose treatment. H9c2 cells were cultured in normal or high glucose DMEM. Arginase activity increased in parallel with increased cell death and oxidative stress. Arginase inhibitor N ω-hydroxy-nor-l-arginine (nor-NOHA) and NOS inhibitor N ω-nitro-l-arginine methyl ester (L-NAME) could reverse these effects. Despite of upregulated NOS activity, NO production was impaired which could be preserved by nor-NOHA, suggesting a decreased substrate availability of NOS due to increased arginase activity. L-arginine supplementation decreased superoxide production while it could not protect cells from death. Upregulated arginase activity in H9c2 treated with high glucose can cause NOS uncoupling and subsequently reactive oxygen species augmentation and cell death. These findings suggest that arginase will be a novel therapeutic target for treatment of diabetic cardiomyopathy.

Enhanced growth inhibition effect of resveratrol incorporated into biodegradable nanoparticles against glioma cells is mediated by the induction of intracellular reactive oxygen species levels.

Resveratrol (Res) has been reported to elicit many cellular responses including cell cycle arrest, differentiation, and apoptosis. This makes it a novel and potential anticancer agent. Moreover, the lipophilicity of Res raises the possibility of its application as a potential model drug in nanoparticle based delivery systems. Resveratrol is incorporated into mPEG-PCL based nanoparticles with high encapsulation efficiency due to its lipophilicity. The significant finding of the current study is that Res-loaded nanoparticles at lower concentration could lead to significantly higher cell death compared to equivalent dose of free Res and this difference of cytotoxicity could not be abrogated by the antioxidant Vitamin E. Furthermore, free Res shows significant less cytotoxicity than the equivalent dose of Res-loaded nanoparticles with the preconditioning of Vitamin E. Meanwhile, reactive oxygen species (ROS) determination revealed the significantly lower intracellular ROS levels in Res-treated cells compared to nanoparticle-treated cells. Therefore, the differential cytotoxicity between Res and Res-loaded nanoparticles may be mediated by the discrepancy of intracellular ROS levels. The present results suggest that Res-loaded nanoparticles could be a potential useful chemotherapeutic formulation for malignant glioma therapy and the development of traditional Chinese medicine with nanoscale drug formation warrants more intensive research in order to evaluate its clinical feasibility.

[Regression of vascular remodeling in renovascular hypertensive rats by tetrandrine and enalapril].

AIM: To observe the regression effect of tetrandrine (Tet) and enalapril (Ena) on vascular morphological changes in renovascular hypertensive (RH) rats. METHODS: Renovascular hypertension was induced by two kidney one clip (2K1C) operation. The morphometric measurements were performed in the aorta, caudal artery, renal arterioles, coronary arterioles and mesenteric arterioles. RESULTS: The wet weight of aorta, caudal artery and femoral artery of RH rats (18 weeks after 2K1C operation) were greater than those of sham-operated rats. The media thickness, lumen diameter, cross section of media, media over lumen ratio and the wet weight of abdomen aorta, caudal artery, coronary arterioles, renal arterioles and mesenteric arterioles were significantly increased, which were more significant in arterioles with the diameter smaller than 70 microns. There were no significant change in the number of the smooth muscle cells (VSMC) in most vessel wall, except in renal arterioles, where the number of smooth muscle cells were significantly increased. After Tet (50 mg.kg-1.d-1, p.o.) or Ena (6 mg.kg-1.d-1, p.o.) treated for 9 weeks from week 9 after 2K1C operation, almost all the changes in the media thickness, the media to lumen ratio, the cross section of media and the wet weight were ameliorated. CONCLUSION: In RH rats, mainly a hypertrophic and rearrangement remodeling in the wall of arteries and arterioles was observed with a proliferation of VSMC in renal arterioles. Tet and Ena were shown to regress vascular remodeling by markedly attenuating these changes in renovascular hypertensive rats.