Effect of Thymoquinone on Renal Damage Induced by Hyperlipidemia in LDL Receptor-Deficient (LDL-R-/-) Mice.

Hyperlipidemia is a well-established risk factor for kidney injury, which can lead to chronic kidney disease (CKD). Thymoquinone (TQ) is one of the most active ingredients in Nigella sativa seeds. It has various beneficial properties, including antioxidant and anti-inflammatory activities. TQ also exerts positive effects on doxorubicin- (DOX-) induced nephropathy and ischemia-reperfusion-induced kidney injury in rats. Therefore, in this study, we investigated the possible protective effects of TQ against kidney injury in low-density lipoprotein receptor-deficient (LDL-R-/-) mice. Eight-week-old male LDL-R-/- mice were randomly divided into the following three groups: normal diet (ND group), high-fat diet (HFD group), and HFD combined with TQ (HFD+TQ group). The mice were fed the same diet for eight weeks. After eight weeks, we performed serological analysis of the mice in all three groups. We histologically analyzed the kidney tissue and also investigated the expression of proinflammatory cytokines in the kidney tissue. Metabolic characteristics, including total cholesterol (TC), low-density lipoprotein-cholesterol (LDL-C), and creatinine (CRE) levels, were lower in the LDL-R-/- HFD+TQ mice than in the HFD mice. Periodic acid-Schiff (PAS) and Masson's trichrome staining revealed excessive lipid deposition and collagen accumulation in the kidneys of the LDL-R-/- HFD mice, which were significantly reduced in the LDL-R-/- HFD+TQ mice. Furthermore, macrophages and levels of proinflammatory cytokines were lower in the kidney tissues of the LDL-R-/- HFD+TQ mice than in those of the LDL-R-/- HFD mice. Moreover, profibrosis- and oxidative stress-related protein expression was lower in the kidney tissues of the LDL-R-/- HFD+TQ mice than in those of the LDL-R-/- HFD mice. These results indicate that TQ may be a potential therapeutic agent for kidney damage caused by hyperlipidemia.

Effects of thymoquinone against angiotensin II­induced cardiac damage in apolipoprotein E­deficient mice.

Herbal medicines have attracted much attention in recent years and are increasingly being used as alternatives to pharmaceutical medicines. Thymoquinone (TQ) is one of the most active ingredients in Nigella sativa seeds, which has several beneficial properties, including anti­inflammatory, anti­oxidative stress, anti­hypertensive, anti­apoptotic and free radical­scavenging effects. Angiotensin II (Ang II) is involved in cardiovascular diseases. The present study aimed to investigate the potential protective effects of TQ against Ang II­induced cardiac damage in apolipoprotein E­deficient (ApoE­/­) mice. Briefly, 8­week­old male ApoE­/­ mice were randomly divided into four groups: Control, TQ, Ang II and Ang II + TQ groups. Osmotic minipumps, filled with either a saline vehicle or an Ang II solution (1,000 ng/kg/min), were implanted in ApoE­/­ mice for up to 4 weeks. The serum levels of high­sensitivity C­reactive protein (hs­CRP) and histopathological alterations in heart tissue were assessed. In addition, the mRNA and protein expression levels of molecules associated with fibrosis (collagen I and III), oxidative stress and apoptosis (Nox4 and p53), and inflammation [tumor necrosis factor (TNF)­α, interleukin (IL)­1ß and IL­6] were analyzed by reverse transcription­quantitative PCR (RT­qPCR) and western blotting. In the in vitro study, H9c2 cells were incubated with different concentrations of Ang II, and the expression levels of pro­inflammatory cytokines were evaluated using RT­qPCR, whereas the protein expression levels of phosphorylated­extracellular signal­regulated kinase (p­ERK) were determined using western blotting. Western blotting was also performed to detect the expression levels of collagen I, collagen III, Nox4 and p53 in H9c2 cells. The results revealed that TQ inhibited the Ang II­induced increases in serum hs­CRP levels. TQ also significantly inhibited the high levels of TNF­α, IL­1ß, IL­6, collagen I, collagen III, Nox4 and p53 in Ang II­treated mice. Furthermore, TQ protected against Ang II­induced cardiac damage by inhibiting inflammatory cell infiltration, proinflammatory cytokine expression, fibrosis, oxidative stress and apoptosis by suppressing activation of the p­ERK signaling pathway. In conclusion, TQ could be considered a potential therapeutic agent for Ang II­induced cardiac damage.

Neuroprotective effect of Trichosanthes kirilowii cassia twig on cerebral ischemia-reperfusion injury in rats.

In this study, in-depth observation and investigation of blood-brain barrier permeability and neuroprotective effect of Trichosanthes kirilowii cassia twig particles on rats with cerebral ischemia-reperfusion injury were performed. Focal cerebral ischemia-reperfusion injury model was established by middle cerebral artery occlusion method, reperfusion was implemented 2 hours after ischemia; qualitative analysis and investigation of trichosanthes kirilowii cassia twig particles in plasma, brain tissue and cerebrospinal fluid in normal and middle cerebral artery occlusion (MCAO) rats were done by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS); changes in neurological deficits, cerebral infarction stereometry, blood-brain barrier permeability and histopathological changes of MCAO model rats were observed. Qualitative analysis by HPLC-MS/MS results showed that ingredients, paeoniflorin, albiflorin, liquiritin in Trichosanthes kirilowii cassia twig particles can reach the brain through the blood-brain barrier. In the model group, glycyrrhizin and glycyrrhizic acid can be detected in brain tissue or cerebrospinal fluid. In addition, Trichosanthes kirilowii cassia twig particles can significantly lower neurological deficits of rats in middle cerebral artery occlusion model, reduce the Evans blue penetration, contract infarct size, and reduce pathological tissue injury of cerebral ischemia reperfusion. The ingredients of Trichosanthes kirilowii cassia twig particles can reach the brain tissue through the blood-brain barrier and play a role in neuroprotection of rats with cerebral ischemia-reperfusion injury, which has important research significance and brings scientific experimental, theoretical basis for clinical drug use.