Astragalin Inhibits Apoptosis of Cerebral Cortex Neurons in APP/PS1 Mice / 中山大学学报(医学科学版)

ObjectiveTo investigate the effect of Astragalin （AST） on apoptosis of cerebral cortex neurons in APP/PS1 transgenic mice. MethodsEighteen six-month-old male APP/PS1 transgenic mice were randomly divided into APP/PS1 group， APP/PS1+ 40 mg/kg AST group and APP/PS1+ 20 mg/kg Donepezil （DNP） group， with six mice in each group. At the same time， six male C57BL/6 mice were selected as the normal control group. After intraperitoneal injection of AST once a day and continuous administration for one month， we used Tunel staining to detect the apoptosis of neurons in the cerebral cortex of APP/PS1 mice； immunofluorescent staining to examine the expression of apoptosis-related proteins Bax， Bcl-2， Caspase9 and Cleaved-Caspase3 in the cerebral cortex neurons of APP/PS1 mice； Western blot method to evaluate the changes of the expression of Bax， Bcl-2， Caspase9 and Caspase3. ResultsTunel staining showed that 40 mg/kg AST and 20 mg/kg DNP both reduced the apoptosis of neurons in the cerebral cortex of APP/PS1 mice， AST with more significant inhibition effect. Immunofluorescent staining revealed that 40 mg/kg AST and 20 mg/kg DNP both inhibited the expression of Bax， Caspase9， and Cleaved-Caspase3， and icreased the expression of Bcl-2 in the cerebral cortex neurons of APP/PS1 mice. Western blot results further confirmed that 40 mg/kg AST and 20 mg/kg DNP both down-regulated the expression of Bax （P < 0.05， P < 0.05）， Caspase9 （P < 0.005， P < 0.05） and Caspase3 （P < 0.0001， P < 0.0001） ， and up-regulated the expresstion of Bcl-2 （P < 0.05， P < 0.05） in the cerebral cortex neurons of APP/PS1 mice. ConclusionsAST can inhibit the apoptosis of cerebral cortex neurons in APP/PS1 mice.

Di'ao Xinxuekang alleviates non-alcoholic steatohepatitis in mice by up-regulating Nrf2/HO-1 signaling pathway / 中国中药杂志

The present study investigated the therapeutic effect and mechanism of Di'ao Xinxuekang(DXXK) on non-alcoholic steatohepatitis(NASH) in mice. Sixty-five C57 BL/6 J mice were randomly divided into a normal group and an experimental group for model induction with the high-fat diet for 16 weeks. Then the mice in the experimental group were randomly divided into a model group, an atorvastatin group(4 mg·kg~(-1)·d~(-1)), and high-(200 mg·kg~(-1)·d~(-1)), medium-(60 mg·kg~(-1)·d~(-1)), and low-dose(20 mg·kg~(-1)·d~(-1)) DXXK groups, with 10 mice in each group. Drugs were administered by gavage for eight weeks. Serum lipid, liver lipid, serum alanine aminotransferase(ALT), aspartate aminotransferase(AST), malondialdehyde(MDA), superoxide dismutase(SOD), and glutathione reductase(GSH-Px) were determined. Interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α) were measured by enzyme-linked immunosorbent assay(ELISA). The liver index was calculated. The liver pathological change and lipid accumulation were observed by HE and oil red O staining. The liver ultrastructure was observed by the transmission electron microscope. The mRNA and protein expression of nuclear factor-erythroid 2 related factor 2(Nrf2) and heme oxygenase-1(HO-1) was detected by real-time fluorescence-based quantitative PCR and Western blot, respectively. The results showed that compared with the normal group, the model group displayed serum lipid and liver lipid metabolism disorders, elevated transaminase, lipid deposition, steatosis, and inflammation, suggesting that the NASH model in mice was properly induced. Compared with the model group, the DXXK groups showed decreased serum lipid, liver lipid, ALT, AST, MDA, IL-1β, and TNF-α, increased SOD and GSH-Px, alleviated hepatic steatosis, ballooning, and inflammation, and up-regulated Nrf2 and HO-1 gene and protein expression. In conclusion, DXXK can significantly alleviate NASH in mice, which is related to the inhibition of oxidative stress and inflammatory damage by up-regulating the Nrf2/HO-1 signaling pathway.

Investigation on Anti-atherosclerosis Mechanism of Di'ao Xinxuekang Based on NLRP3 Inflammasome / 中国实验方剂学杂志

Objective:To investigate the effects of Di'ao Xinxuekang （DXXK） on NLRP3 inflammasome in mouse RAW264.7 macrophages and thoracic aorta of rats with atherosclerosis （AS）， so as to explore its anti-AS mechanism. Method:RAW264.7 cells were stimulated with oxidized low density lipoprotein （ox-LDL） and then intervened with MCC950 and DXXK. The contents of tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>） and interleukin-1<italic>β</italic> （IL-1<italic>β</italic>） were determined by enzyme linked immunosorbent assay （ELISA）. The mRNA and protein expression levels of Nod-like receptor protein 3 （NLRP3）， inflammasome adaptor protein apoptosis-associated speck-like protein containing CARD （ASC）， and cysteine-dependent aspartate-directed protease-1 （Caspase-1） were detected by real-time polymerase chain reaction （Real-time PCR） and Western blotting. Sixty male SD rats were randomly divided into the normal group， model group， atorvastatin group （2.0 mg·kg^-1）， as well as high-， medium-， and low-dose （100， 30， and 10 mg·kg^-1） DXKK groups， with 10 rats in each group. The rats were exposed to the high-fat diet and vitamin D₂ for inducing AS. The blood lipid level was measured using an automatic biochemical analyzer， followed by the calculation of AS index （AI）. The contents of serum TNF-<italic>α</italic> and IL-1<italic>β</italic> were determined by ELISA， and the mRNA and protein expression levels of NLRP3， ASC， and Caspase-1 in thoracic aorta were assayed by Real-time PCR and Western blotting. HE staining and Sirius red staining were conducted to observe the pathomorphological changes in the abdominal aorta and aortic sinus. Result:Compared with the normal group， the model group exhibited significantly increased TNF-<italic>α</italic> and IL-1<italic>β</italic> contents and up-regulated NLRP3， ASC， and Caspase-1 mRNA and protein expression in RAW264.7 cells （<italic>P</italic><0.01）. The above indexes in each drug administration group were significantly reduced in contrast to those in the model group （<italic>P</italic><0.05， <italic>P</italic><0.01）. The comparison with the model group showed that cholesterol （TC）， triglyceride （TG）， low-density lipoprotein cholesterol （LDL-C）， and AI in each DXXK group significantly declined， while the high-density lipoprotein cholesterol （HDL-C） was significantly elevated （<italic>P</italic><0.05， <italic>P</italic><0.01）. The levels of serum TNF-<italic>α</italic> and IL-1<italic>β</italic> and the mRNA and protein expression levels of NLRP3， ASC， and Caspase-1 in the thoracic aorta were decreased （<italic>P</italic><0.05， <italic>P</italic><0.01）. Abdominal aortic lesions and fibrous hyperplasia of aortic sinus were significantly improved. Conclusion:DXXK has a significant anti-AS effect， which is possibly related to the inhibition of NLRP3 inflammasome.

Effects of volatile oil from Acori Graminei Rhizoma on glial fibrillary acidic protein and immediate early genes expressions in the basal lateral amygdala of the inflammatory pain rats / 解剖学报

Objective To construct a rat model of inflammatory pain by injecting complete Freund' adjuvant (CFA) to study effects of volatile oil of Acori Graminei Rhizoma on the expression of glial fibrillary acidic protein (GFAP) and immediate early gene c-fos in the basal lateral amygdale (BLA) of the inflammatory pain rats. Methods Thirty-six adult male SD rats were randomly divided into 6 groups; control group, sham group, CFA group, CFA+ 5 g/( kg · day) volatile oil of Acori Graminei Rhizoma group, CFA+10 g/(kg · day) volatile oil of Acori Graminei Rhizoma group, CFA+20 g/(kg · day) volatile oil of Acori Graminei Rhizoma group, six rats in each group were taken gavage for 21 days. Immunofluorescence and Western blotting methods were used to detect the expressions of GFAP and c-fos in the BLA of all rats. Results Immunofluorescence and Western blotting results showed that compared with the control group, the positive expression of GFAP and c-fos in the BLA of the CFA rats were significantly increased (P<0.01). After treatment of the volatile oil from Acori Graminei Rhizoma, the positive expressions of GFAP and c-fos were reduced compared to the CFA group, as well as the expression levels were decreased in the dose-dependent manner (P<0.01). Compared with the low dose group, the positive expression of GFAP and c-fos of high dose group were decreased significantly (P<0.01). Conclusion The volatile oil fraction from Acori Graminei Rhizoma could reduce the expressions of GFAP and c-fos the BLA of CFA-induced chronic inflammatory pain model rats.

Effect of Total Saponin of Dioscoreae Collettii Rhizoma on Toll-like Receptor/Nuclear Factor-κB (TLR/NF-κB) Signaling Pathway Induced by Monosodium Urate in THP-1 Cells / 中国实验方剂学杂志

Objective::To investigate the effect of total saponin of Dioscoreae Collettii Rhizoma (TSD) on Toll-like receptor/nuclear factor-κB (TLR/NF-κB) signaling pathway induced by monosodium urate in THP-1 cells, in order to explore the possible mechanism of anti-gout arthritis. Method::Phorbol 12-myristate 13-acetate (PMA)-induced THP-1 cells were differentiated into macrophages, divided into normal group, model group, low, medium and high-concentration TSD groups (1, 3, 10 mg·L-1) and colchicine group (0.2 mg·L-1). Except the normal group, the other groups were stimulated with 400 mg·L-1 monosodium urate to replicate an inflammation model in vitro. Cell viability was measured by methyl thiazolyl tetrazolium (MTT) assay, the levels of inflammatory factors tumor necrosis factor-α(TNF-α ) and interleukin-1β(IL-1β) were detected by enzyme-linked immunosorbent assay (ELISA). The protein levels of Toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88) and NF-κB were detected by Western blot. The mRNA levels of TLR4, NF-κB and Pro-IL-1β were measured by real-time fluorescence quantitative PCR (Real-time PCR), and the nuclear shift of NF-κB p65 was detected by immunofluorescence. Result::0~32 mg·L-1 TSD has no effect on cell viability. Compared with the normal group, the secretion levels of inflammatory factors TNF-α and IL-1β in the model group were significantly increased (P<0.01), and the expressions of key proteins (TLR4, MyD88 and NF-κB) and genes (TLR4, NF-κB and Pro-IL-1β) were increased (P<0.01). Compared with the model group, 1-30 mg·L-1 TSD significantly down-regulated the secretion of inflammatory factors TNF-α and IL-1β (P<0.01), the expressions of key proteins (TLR4, MyD88 and NF-κB) and genes (TLR4, NF-κB and Pro-IL-1β) were decreased (P<0.05, P<0.01), and the NF-κB p65 partially trans-located to the cytosol and the superposition in the nucleus were decreased, inhibiting the nuclear translocation of NF-κB p65. Conclusion::TSD may exert an anti-inflammatory effect by down-regulating the expressions of TLR4, NF-κB and Pro-IL-1β mRNA and reducing the secretion of inflammatory factors TNF-α and IL-1β.

Regulatory effect of Di'ao Xinxuekang on TLR4/MyD88/NF-κB signaling pathway in atherosclerotic rats / 中国中药杂志

The aim of this paper was to observe the effect of Di'ao Xinxuekang(DXXK) on TLR4/MyD88/NF-κB signaling pathway in atherosclerotic rats, and to explore its anti-atherosclerotic mechanism. Sixty SD rats were randomly divided into normal group, model group, atorvastatin group(4.0 mg·kg~(-1)), and DXXK groups(100, 30, 10 mg·kg~(-1)), with 10 rats in each group. The atherosclerosis model was induced by high fat diet plus vitamin D_2. Experimental drugs were administered intragastrically once daily for 8 weeks starting from the 9 th week. Biochemical analyzers were used to detect levels of triglyceride(TG), total cholesterol(TC), low-density lipoprotein cholesterol(LDL-C) and high-density lipoprotein cholesterol(HDL-C) in blood lipid. The levels of serum tumor necrosis factor(TNF)-α, interleukin(IL)-6 and IL-1β were detected by ELISA. Pathological changes of aortic tissues were observed by using Sudan Ⅳ and HE staining. The mRNA and protein expressions of TLR4, MyD88 and NF-κB p65 in aortic tissues were detected by RT-PCR and Western blot, respectively. As compared with the model group, TC, TG, and LDL-C levels in serum were significantly decreased, HDL-C content was significantly increased, and levels of TNF-α, IL-6, and IL-1β in serum were significantly decreased in atorvastatin group and DXXK high and middle dose groups. Aortic lesions in atorvastatin group and DXXK group were significantly improved, and the mRNA and protein expressions of TLR4, MyD88, NF-κB p65 in the aorta were decreased. DXXK has a preventive and therapeutic effect on atherosclerosis in rats, and its mechanism may be related to inhibiting inflammatory reaction by regulating TLR4/MyD88/NF-κB signal transduction, thereby inhibiting the progression of atherosclerosis.

Evaluation of Anterior Ethmoidal Artery by 320-Slice CT Angiography with Comparison to Three-Dimensional Spin Digital Subtraction Angiography: Initial Experiences

OBJECTIVE: To explore the usefulness of 320-slice CT angiography (CTA) for evaluating the course of the anterior ethmoidal artery (AEA) and its relationship with adjacent structures by using three-dimensional (3D) spin digital subtraction angiography (DSA) as standard reference. MATERIALS AND METHODS: From December 2008 to December 2010, 32 patients with cerebrovascular disease, who underwent both cranial 3D spin DSA and 320-slice CTA within a 30 day period from each other, were retrospectively reviewed. AEA course in ethmoid was analyzed in DSA and CTA. In addition, adjacent bony landmarks (bony notch in medial orbital wall, anterior ethmoidal canal, and anterior ethmoidal sulcus) were evaluated with CTA using the MPR technique oriented along the axial, coronal and oblique coronal planes in all patients. The dose length product (DLP) for CTA and the dose-area product (DAP) for 3D spin DSA were recorded. Effective dose (ED) was calculated. RESULTS: The entire course of the AEA was seen in all 32 cases (100%) with 3D spine DSA and in 29 of 32 cases (90.1%) with 320-slice CTA, with no significant difference (p = 0.24). In three cases where AEA was not visualized on 320-slice CTA, two were due to the dominant posterior ethmoidal artery, while the remaining case was due to diminutive AEA. On MPR images of 320-slice CT, a bony notch in the orbital medial walls was detected in all cases (100%, 64 of 64); anterior ethmoidal canal was seen in 28 of 64 cases (43.8%), and the anterior ethmoidal sulcus was seen in 63 of 64 cases (98.4%). The mean effective dose in CTA was 0.6 +/- 0.25 mSv, which was significantly lower than for 3D spin DSA (1.3 +/- 0.01 mSv) (p < 0.001). CONCLUSION: 320-slice CTA has a similar detection rate for AEA to that of 3D spin DSA; however, it is noninvasive, and may be preferentially used for the evaluation of AEA and its adjacent bony variations and pathologic changes in preoperative patients with paranasal sinus diseases.