Differential Expression and Signal Transduction Analysis of MicroRNA Related to Kidney Yang Deficiency in Kidney Tissues of Rats / 中国实验方剂学杂志

Objective: Microarray chip was used to detect the differentially expressed microRNA (miRNA) in kidney tissues of rats with kidney-Yang deficiency induced by adenine,and its significance was analyzed by bioinformatics method. Method: Rats with kidney-Yang deficiency were induced by intragastric administration of 150 mg·kg-1 adenine in model group, while rats in normal group were given the same amount of saline.Kidney tissues were taken for hematoxylin-eosin (HE) staining pathological sections after anesthesia and blood urea nitrogen (BUN), creatinine (SCr) in blood and 24-hour urinary protein (24U-TP) in urine were measured. μParaflo microfluidic chip technology was used to investigate differential expression miRNA in kidney tissues, and microarray results were verified by Real-time PCR. Bioinformatics database was used to analyze the target genes and functions of differential expression miRNAs. Result: Gene chip results showed that there were 50 differentially expressed microRNAs after modeling. Compared with control group, only 9 miRNAs were highly expressed in kidney tissues with significant difference were detected in model group. Compared with the normal group, the expression of rno-miR-21-5p, rno-let-7i-5p, rno-miR-146b-5p and rno-miR-15b-5p in model group increased significantly(PPPPPConclusion: This experiment found 4 miRNAs involved in the regulation of renal interstitial fibrosis (RIF) and 2miRNAs with unknown functions, which provided a new clue for further analysis of the regulatory network of kidney-yang deficiency.

Effect of Zhenwu Tang on regulating of "AVP-V2R-AQP2" pathway in NRK-52E cells / 中国中药杂志

This study was aimed to investigate the effect and mechanism of Zhenwu Tang on AVP-V2R-AQP2 pathway in NRK-52E cells . Forty eight male SD rats were randomly divided into eight groups with 6 animals in each group. Distilled water or 22.68 g·kg⁻¹·d⁻¹ Zhenwu Tang(calculated by raw drug dosage meter) was given by gavage. Blood samples were collected by cardiac puncture， and the medicated serum was centrifuged from the blood by 3 000 r·min⁻¹. NRK-52E cells were treated with different medicated serum or dDAVP. The condition of cell proliferation was detected by RTCA. The distribution of V2R and AQP2 in cells were detected by immunofluorescence. The expression of V2R， PKA and AQP2 were detected by Western blot and AQP2 mRNA level was detected by real-time PCR. Results showed that the level of AQP2 mRNA(<0.01) and protein expression of V2R， PKA and AQP2(<0.05， <0.01， <0.05) of Z7d group which was treated with Zhenwu Tang medicated serum for 24 h were significantly higher than that of normal rat serum group. And the expression level of V2R， p-AQP2 and AQP2(<0.01， <0.05， <0.01) of Z7d+dDAVP group were significantly increased comparing to normal rat serum group. The results indicate that the applying of Zhenwu Tang medicated serum could increase the expression level of V2R， PKA and AQP2 which exist in AVP-V2R-AQP2 pathway in NRK-52E， and there is synergistic effect between Zhenwu Tang medicated serum and dDAVP. So the pathway of AVP-V2R-AQP2 may be one of the mechanism for which Zhenwu Tang regulate balance of water transportation.

Protective effect of β-asarone on AD rat model induced by intracerebroventricular injection of Aβ₁₋₄₂ combined 2-VO and its mechanism / 中国中药杂志

This study was aimed to investigate the protective effect and mechanism of β-asarone on the animal model of Alzheimer's disease(AD) which was induced by intracerebroventricular injection of Aβ₁₋₄₂ combined cerebral ischemia. One hundred and five rats were randomly divided into seven groups including sham-operated group, AD model group, β-asarone 10 mg•kg⁻¹ group, β-asarone 20 mg•kg⁻¹ group, β-asarone 30 mg•kg⁻¹ group, donepezil group(0.75 mg•kg⁻¹) and Ginkgo biloba extract group(24 mg•kg⁻¹). Rats' learning and memory abilities, cerebric regional blood flow, pathological changes in hippocampal CA1 region, the expression level of HIF-1α and serum CAT, SOD and MDA level were detected 4 weeks later. The results showed that the application of intracerebroventricular injection of Aβ₁₋₄₂ joint 2-VO could lead to rats' dysfunction of learning and memory, decrease in regional cerebral blood flow. Neurons in CA1 region were arranged in disorder, and amyloid deposition was increased. The number of cerebral cortical cells expressing HIF-1α was increased as well. The level of serum CAT and SOD decreased, while level of serum MDA increased. However these symptoms were improved by 20 mg•kg⁻¹ and 30 mg•kg⁻¹ β-asarone. The results indicated that β-asarone could effectively relieve the symptoms of the AD model induced by intracerebroventricular injection of Aβ₁₋₄₂ combined cerebral ischemia, and the potential mechanism might be that it could attenuate damage of MDA to the body by improving the level of CAT and SOD, meanwhile the level of HIF-1α decreased as the decline of hyperoxide which might attenuate its damage to neuron, so it finally achieved alleviating Alzheimer's disease.

Protective effect of β-asarone on PC12 cells injury induced by Aβ₁₋₄₂ astrocytic activation / 中国中药杂志

This study was aimed to investigate the protective effect and mechanism of β-asarone on PC12 cells injury induced byAβ₁₋₄₂ activated astrocytes, and provide experimental basis for β-asarone application in the prevention and control of Alzheimer's disease (AD). Firstly, RA-h and PC12 cells were co-cultured in the special transwell chamber, and the Real time cell analysis (RTCA) system was used to real-time observe its effect on PC12 cells survival rate in the co-culture system after astrocytes injury induced by Aβ₁₋₄₂. The best intervention time of β-asarone was selected according to the survival curve and parameters generated automatically. β-asarone with different concentrations was used for intervention on astrocytes, then the changes of PC12 cells survival rate in the co-culture system were observed. Secondly, MTT assay was used to detect the effect of Aβ₁₋₄₂ on PC12 cells survival rate as well as the intervention effect of β-asarone, and verify the testing results of RTCA. The levels of IL-1β, TNF-α and BDNF in culture media of the lower chamber were detected by ELISA. The NF-κB activity and phosphorylation levels of ERK, p38 and JNK were detected by Western blot. Results showed that β-asarone (55.5 mg•L⁻¹) could significantly slowdown the decline of PC12 cells survival rate caused by Aβ₁₋₄₂-induced RA-h activation (P<0.01), significantly reduce the levels of IL-1β, TNF-α and the phosphorylation levels of ERK, p38 and JNK in culture media of the lower chamber (P<0.01). β-asarone(166.7 mg•L⁻¹) could promote the release of BDNF in culture media of the lower chamber(P<0.05). These results indicated that Aβ₁₋₄₂ could induce RA-h activation and its release of IL-1β, TNF-α and other inflammatory factors to aggravate the PC12 cells injury; β-asarone could reduce the levels of IL-1β, TNF-α, promote the release of BDNF, and inhibit the NF-κB activity as well as phosphorylation levels of ERK, p38 and JNK protein in PC12 cells.

Adverse drug reactions induced by Mailuoning injection: literature review on 162 cases / 中国中药杂志

To understand the adverse drug reaction (ADR) induced by Mailuoning injection. 162 ADRs due to the drug were retrieved from national medical journals of 1988-2007 for statistics. It was shown that there was no relationship between ADR and dosage, but ADR appeared mostly in middle-aged and old groups, and more in male than in female. The occurrence of ADR was commonly within 30 min after injection. It involved injuries of various systems and organs. As for the 123 cases of allergy, 38 were anaphylactic shock (accounting for 23.46%), two people died. ADR characterized by immediate type on initial use and tachy type. It should be paid more attention to the Mailuoning injection for the immediate hypersensitivity reaction (such as anaphylactic shock).

Effect of 2-methoxycinnamaldehyde on activity of COX and PGE2 release in cerebral microvascular endothelial cells stimulated by IL-1 / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the effect of 2-methoxycinnamaldehyde (isolated from fraction A of Guizhi Tang) on activity of COX and PGE2 release in rat cerebral microvascular endothelial cells (rCMEC) stimulated by IL-1.</p><p><b>METHOD</b>rCMEC were cultured, and identified by immunohistochemistry for von Willebrand factor (VIII factor, a marker for all endothelial cells) in cytoplasm of the cells. Different concentrations of 2-methoxycinnamaldehyde were added respectively and incubated for 3 hours, then stimulated for another 12 hours by IL-1. Activities of COX-1 and COX-2 in rCMEC, and production of PGE2 in the conditioned media were measured by ELISA.</p><p><b>RESULT</b>Positive immunostaining for VIII factor was present diffusely in the cytoplasm of > 90% rCMEC. After being exposed to 30 ng x mL(-1) IL, the activity of COX-2 in rCMEC and the production of PGE2 in conditioned media were higher than those of control group, while there was no difference on activity of COX-1 in the two groups. 2-methoxycinnamaldehyde could down-regulate them in concentration-dependently, and significant differences on the activity of COX-2 and amount of PGE2 were showed in 200 microg x mL(-1) concentration.</p><p><b>CONCLUSION</b>2-methoxycinnamaldehyde can affect the PGE2 release in rCMEC induced by IL-1, which might be related with its inhibition on the activity of COX-2.</p>