Tongqiao Huoxue Decoction Research Articles

Effect of Tongqiao Huoxue Decoction on cAMP-PKA Signaling Pathway in Rats with Vascular Dementia

Effect of Tongqiao Huoxue Decoction on cAMP-PKA Signaling Pathway in Rats with Vascular Dementia

Tongqiao Huoxue Decoction ameliorates learning and memory defects in rats with vascular dementia by up-regulating the Ca2+-CaMKII-CREB pathway

The present study was aimed at determining the effects of Tongqiao Huoxue Decoction (TQHXD) on the Ca2+-CaMKII-CREB pathway and the memory and learning capacities of rats with vascular dementia (VD). The rat VD model was established by using an improved bilateral carotid artery ligation method. The Morris water maze experiment was used to evaluate the ethology of the VD rats following treatments with TQHXD at 3.01, 6.02, and 12.04 g·kg−1 per day for 31 days. At the end of experiment, the hippocampus were harvested and analyzed. Western blotting and RT-PCR were used to measure the expression levels of calmodulin-binding protein kinase II(CaMKII), protein kinase A(PKA), cAMP-response element binding protein(CREB), and three N-methyl-D-aspartic acid receptor subunits (NR1, NR2A, and NR2B). Our results revealed that TQHXD could alleviate the loss of learning abilities and increase the memory capacity (P < 0.05 and P < 0.01 vs the model group, respectively). The treatment with 6.02 and 12.04 g·kg−1 of TQHXD significantly up-regulated the Ca2+-CaMKII-CREB pathway in the hippocampus. In conclusion, TQHXD showed therapeutic effects on a bilateral carotid artery ligation-induced vascular dementia model, through the up-regulation of calcium signalling pathways.

Effects of Muscone on the Expression of P-gp, MMP-9 on Blood-Brain Barrier Model In Vitro.

Muscone is the main chemical ingredient in Musk which is main crude drug in Tongqiaohuoxue decoction (TQHXD), and TQHXD has a protective effect on damaged neurons, so we hypothesize that muscone can alter blood-brain barrier (BBB) permeability via the modulation of P-glycoprotein (P-gp) and matrix metalloproteinase-9 (MMP-9) expression. In this study, astrocytes (AC) and human umbilical vein endothelial cells (ECV304) were co-cultured to simulate the BBB model in vitro. Leak testing, transmembrane resistance experiments, and BBB-specific enzyme testing were used to test whether the model was successful. Different concentrations of muscone permeating the BBB were detected by gas chromatography (GC). The change of the transendothelial electrical resistance (TEER) on the BBB in vitro after treating with muscone was detected by Millicell-ERS. The protein expression of P-gp, MMP-9 in normal, and oxygen/glucose deprivation (OGD) BBB model was determined by western blotting to inquire that the mechanism of muscone penetrates the BBB model in vitro. The results show that muscone was detected in the lower medium of the BBB model by GC; the values of TEER were no significant difference before and after muscone (8 μM) was added to the BBB model; the expression of P-gp significantly decreased after the BBB model treatment with muscone (4, 8, and 16 μM) for 24 h; the expression of P-gp and MMP-9 in different concentrations of muscone groups had different degrees of reduction compared with the BBB in the state of OGD. In conclusion, muscone could permeate the BBB model, and it was associated with the inhibition of P-gp and MMP-9 expression. An understanding of the mechanisms of muscone across the BBB is crucial to the development of therapeutic modalities for cerebral vascular diseases.

Clinical Evidence of Chinese Herbal Medicine for Treatment of Idiopathic Sudden Sensorineural Hearing Loss from Chinese Literature

Purpose: To provide a comprehensive summary of all clinical evidence on Chinese herbal medicine (CHM) for idiopathic sudden sensorineural hearing loss (ISSHL) published in Chinese literature. Methods: We systematically searched randomized clinical trials (RCTs), clinical controlled trials (CCTs), case series (CSs) and case reports (CRs) which reported CHM for ISSHL through four main Chinese electronic databases from their inception to March 2014. We bibliometrically analyzed the studies and assessed themethodological quality of RCTsusing the Cochrane risk of bias tool. Results: A total of 299 clinical studies with involving 22,237 participants were identified including 150 RCTs, 42 CCTs, 80 CSs and 27 CRs. The number of publications increased obviously per year from 1995, with the peak in 2011. Among 145 different herbal formulae tested, the most popular prescribed herbal formulae were Longdan Xiegan decoction and TongqiaoHuoxue decoction, and the top three frequently used Chinese herbs were Rhizoma Chuanxiong, Radix Bupleuri and Radix Puerariae Lobatae. The most frequently reported outcome was improvement of hearing in 286 (95.7%), followed by improvement of tinnitus (139, 46.5%), improvement of dizziness (99, 33.1%). Among the 150 RCTs, randomizationmethods were described in only 12 trials (8.0%). No trial reported allocation concealment and only four mentioned blinding. Among 146 RCTs (97.3%) and 37 CCTs (88.1%) reporting improvement of hearing as the outcome measurement, all showed significant difference favoring CHM. Of 16 trials reporting adverse events, only five trials reported mild adverse events related to CHM and the remaining stated that none had occurred. Conclusion: The quantity of clinical research on CHM for ISSHL is substantial, but methodological quality of RCTs is generally suboptimal. Future clinical studies would need to report structurally and based on the CONSORT and TREND Statements. Quality of life, adverse events, depression and anxiety should be addressed as outcome measures. Contact: Chun-Xiang Su, susu18182004@126.com Corresponding Author: Jian-Ping Liu (Center for Evidence-Based Chinese Medicine, Beijing University of Chinese Medicine, Chaoyang District, Beijing 100029, China), jianping l@hotmail.com

Effect of Tongqiao-Huoxue decoction on hippocampus neuron apoptosis and blood serum nidogen of vascular dementia rats

Objective To study the effects of Tongqiao-Huoxue decoction on neuron apoptosis in the CA1 region of the hippocampus and the serum nidogen expression of vascular dementia（VaD） rats and to investigate its mechanism. Methods 40 healthy male SD rats were randomly divided into a normal group, a sham operation group, a model group, a TCM group, and a placebo group. VaD rat models were established by Olsson method. After the models were successfully prepared, the rats in the TCM group were fed Tongqiao-Huoxue decoction, the rats in the placebo group were fed distilled water. Morris water maze test was adopted to investigate their learning and memory abilities, TUNEL was used to assay hippocampal CA1 neurons apoptosis, ELISA kits was used to detect Nestin serum levels . Results The neuron apoptosis index in the CA1 region of the hippocampus in the TCM group was（37.01±2.23）%,which was（55.15±1.54）%in the placebo group;the difference was statistically significant（P〈0.05）.The serum nidogen was（4.47±0.31）ng/L in the TCM group, which was（3.42±0.43）ng/L in the placebo group;the difference was statistically significant（P〈0.05）. Compared to the VaD group, escape latency in the TCM group in the third, forth, and fifth day was significantly shorter（P〈0.05）, the number of cross platforms were significantly increased（t=3.521, P=0.000）. Conclusion Tong qiao-Huoxue decoction can improve learning and memory impairment in VaD rats, which may be associated with its increasing serum nidogen expression and decreasing the neuron apoptosis in hippocampal CA1. Key words: Vascular dementia; Nidogen; Neuronal apoptosis; Spatial memory

Serum containing Tongqiaohuoxue decoction suppresses glutamate-induced PC12 cell injury.

Glutamate application is an established method of inducing PC12 cell injury. PC12 cells were cultured with serum containing Tongqiaohuoxue decoction consisting of moschus, Carthamus tinctorius, Rhizoma chuanxiong, Semen pruni persicae, and Radix Paeoniae Rubra. After 24 hours of co-cultivation, glutamate (12.5 mM) was added to the culture medium. We found that serum containing Tongqiaohuoxue decoction prevented the increase in reactive oxygen species, and the decreases in superoxide dismutase and Na+-K+-ATPase activity, induced by glutamate. It also reduced the concentration of malondialdehyde, enhanced the mitochondrial transmembrane potential, inhibited the elevation of cellular calcium, and decreased phosphorylation of calmodulin-dependent protein kinase II. Thus, serum containing Tongqiaohuoxue decoction had protective effects on cell proliferation and membrane permeability in glutamate-injured PC12 cells.

Neuroprotective effects of serum with Tongqiao Huoxue decoction (TQHXD)against glutamate-induced neurotoxicity in PC12 cells

To observe the protective effect of serum with Tongqiao Huoxue decoction (TQHXD) on PC12 cells damaged by glutamate(Glu) and provide clinical proof of the formulae. Sprague Dawley rats underwent intragastric administration of Tongqiao Huoxue decoction twice a day for five days, the administration dose for rats was 4 g x kg(-1). Thus the serum containing TQHXD was prepared. The model of neurocytes damaged by Glu had been established by adding 12.5 mmol x L(-1) Glu to culture medium of PC12 cells. Photic microscope had been used to observe the morphologic changes of cells, the proliferation and activity of PC12 cells had been detected by MTT. Cell membrane permeability had been investigated by the methods of the coloration of trypan blue and AO/EB, the activities of LDH and the contents of NO in the supernatant of PC12 cells had been detected after 5%, 10%, 20% TQHXD being added to the culture medium of PC12 cells damaged by Glu. The morphologic changes in cells were observed under the inverted microscope. Normal PC12 cells in control group were full and bright, but the cells exposed to Glu were shrank; moreover, some cells were disrupted, configuration of the cells in the TQHXD group was closed to that of normal group. Compared with model group, the ratio of living cells in the group being treated by TQHXD after trypan blue staining had a significant increase. After AO/EB staining, cells in TQHXD groups were less being stained by EB. The absorbance (A) values of TQHXD group increased largely. LDH and NO in cell culture supernatant had decreased, it had shown the marked difference. TQHXD showed an apparent protective effect on neurocytes damaged by Glu on proliferation activity and membrane permeability.