Research progress on in vitro models of cardiomyocyte injury / 中国中药杂志

Cardiovascular diseases seriously endanger human health and life. The accompanying myocardial injury has been a focus of attention in society. Chinese medicine,serving as a natural and precious reservoir for the research and development of new drugs,is advantageous in resisting myocardial injury due to its multi-component,multi-pathway,and multi-target characteristics. In recent years,with the extensive application of culture method for isolated cardiomyocytes,a cost-effective,controllable in vitro model of cardiomyocyte injury with uniform samples is becoming a key tool for mechanism research on cardiomyocyte injury and drug development.A good in vitro model can reduce experimental and manpower cost,and also accurately stimulate clinical changes to reveal the mechanism. Therefore,the selection and establishment of in vitro model are crucial for the in-depth research. This study summarized the modeling principles,evaluation indicators,and application of more than ten models reflecting different clinical conditions,such as injuries induced by hypoxia-reoxygenation,hypertrophy,oxidative stress,inflammation,internal environmental disturbance,and toxicity. Furthermore,we analyzed advantages and technical difficulties,aiming to provide a reference for in-depth research on myocardial injury mechanism and drug development.

Research progress in building and applying cardiomyocytes injury model / 中南大学学报(医学版)

Cardiomyocytes injury model has been widely used in the study for the molecular mechanism of cardiovascular diseases and drug action. It is very important to select the appropriate model due to the different formation mechanisms for various models. Clinical cardiovascular pathological change is relatively complex. Currently used models according to the characteristics of clinical cardiovascular diseases mainly include hydrogen peroxide-induced myocardial cell damage model, hypoxia reoxygenation injury model, adriamycin-induced myocardial cell damage model, high sugar high fat-induced myocardial cell damage model, and isoprenaline-induced myocardial cell damage model. Every model has its advantages as well as its disadvantages. The suitable model of myocardial cell injury can be selected according to the research purpose.

Effects of enteral ecological nutrition on intestinal immune function in intestinal mucosa of rats with small intestinal injury / 中国中西医结合急救杂志

Objective To investigate the effects of enteral ecological nutrition on intestinal immune function and Hh protein expression in intestinal mucosa of rats with small intestinal injury and explore the mechanism. Methods Thirty male clean grade Wistar rats were selected as study subjects. The rats were randomly divided into a model group and an enteral ecotrophic group with 15 rats in each group. The small intestinal injury model was prepared by trauma method. After successful modeling, Six hours after successful establishment of the model, the rats in the two groups were fed with 753.12 kJ·kg-1·d-1 energy, 3 times a day. The rats in the model group were fed with conventional diet, while the rats in the enteral ecotrophic group were fed with enteral nutrition emulsion +Lactobacillus, bifidobacterium triple viable bacteria by intragastric administration of 1×107 cfu/d. After 10 days, the rats were killed, the small intestine of the two groups was dissected and stained with hematoxylin-eosin (HE) staining, and the morphological changes of small intestinal mucosa (villus height, glandular recess depth, mucosal thickness) were observed in the two groups; the expressions of CD3+, CD4+, CD8+ positive T cells in small intestinal mucosa were evaluated by immunohistochemistry; the expression of Hh protein in small intestinal mucosa was detected by Western Blot in the two groups. Results On the first day after the establishment of the model, the weight of rats in both groups was lower than that before the modeling [model group (g): 118.0±4.2 vs. 121.7±5.2, enteral ecotrophic group (g):117.5±4.7 vs. 120.8±5.0, P > 0.05], from the fifth day after the modeling, the weight of the rats in the enteral ecotrophic group was significantly higher than that of the model group (g: 127.1±5.0 vs. 123.2±4.2, P < 0.05), continued to 10 days (g: 142.5±6.6 vs. 135.3±6.2, P < 0.05). After the establishment of the model for 10 days, the small intestinal villus height, glandular recess depth, mucosal thickness and percentages of CD3+, CD4+ and CD8+ positive T cells in enteral ecotrophic group were significantly higher than those in model group [villus height (μm): 221.7±25.0 vs. 159.5±20.8, glandular recess depth (μm): 79.39±12.65 vs. 67.87±7.79, mucosal thickness (μm): 254.7±51.8 vs. 209.0±27.2, CD3+: 0.193±0.035 vs. 0.125±0.031, CD4+: 0.130±0.027 vs. 0.104±0.015, CD8+: 0.165±0.026 vs. 0.137±0.027, all P < 0.05]. The expression of Hh protein in the enteral ecotrophic group was obviously higher than that in model group (Hh/β-actin: 0.16±0.04 vs. 0.04±0.02, P <0.05). Conclusion Enteral ecological nutrition may promote the repair of intestinal mucosa and the improvement of immune function level by enhancing the expression of Hh protein in small intestinal mucosa of rats with small intestinal injury.

Effect of sciatic nerve pulsed radiofrequency glial activation levels in spinal dorsal horn in chronic constriction injury rat models and its analgesia effect / 吉林大学学报(医学版)

Objective:To observe the effect of sciatic nerve pulsed radiofrequency (PRF) on the activation levels of microglia cells and astrocytes in spinal dorsal horn in the chronic constriction injury (CCI) rat models, and to explore the relationships between the analgesic mechanism of sciatic nerve PRF and the activation levels of microglia cells and astrocytes in the spinal dorsal horn.Methods:Forty male SD rats were randomly divided into CCI shamoperation+RPF sham group (SS group) , CCI sham-operation+RPF group (SP group) , CCI+RPF sham group (CS group) , CCI+RPF group (CP group) (n=10) .PRF was applied on the sciatic nerve on the 4th day after CCI operation for 120s, with a maximum temperature of 42℃.The mechanical withdrawal thresholds (MWT) and thermal paw withdrawal latencies (TWL) were measured to evaluate mechanical hyperalgesia and thermal hyperalgesia 1dbefore operation (D0) and 1, 3, 5, 7dafter operation (D1, D3, D5, D7) .Western blotting method was used to detect the protein expression levels of ionized calcium binding adapter molecule 1 (Iba-1) and glial fibrillary acidic protein (GFAP) in the ipsilateral spinal dorsal horn of L3-5after pain behavioral test (D7) .Results:Compared with SS group, the rats in CS group after CCI had the significant behavioral changes, such as hallux valgus, lameness, toe bending, and foot raising during walking;MWT and TWL were decreased significantly (P＜0.01) ;the expression levels of Iba-1and GFAP proteins in the ipsilateral spinal dorsal horn were increased significantly (P＜0.05) .Compared with CS group, the behavioral changes of the rats in CP group (D5, D7) such as hallux valgus, lameness, toe bending, and foot raising during walking were alleviated significantly;MWT and TWL were increased significantly (P＜0.01) ;the Iba-1protein expression level in spinal dorsal horn was downregulated significantly (P＜0.05) and the GFAP protein expression level did not change significantly (P＞0.05) .Conclusion:PRF on sciatic nerve can relieve the neuropathic pain (NP) of the CCI rat models;PRF on sciatic nerve can inhibit activation of the microglia cells in the spinal dorsal horn.The analgesic mechanism of PRF on sciatic nerve may be associated with the inhibition of the activation of microglia cells in the spinal dorsal horn.

Studies on characteristics of liver injury model induced by rifampicin and isoniazid in mice / 中国药理学通报

Aim: To establish an ideal anti-tuberculosis drug induced liver injury model and provide a suitable animal model for its pharmacodynamic evaluation. Methods To explore the contribution rate and interaction of isoniazid (INH) and rifampicin (RIF) on liver injury, mice received intragastric administration of RIF 100 nig · kg-1, 300 mg · kg-1 once, or RIF 300 mg · kg-1, INH 150 mg · kg-1, and RIF 300 mg · kg-1 + INH 150 mg · kg-1 for 1 -3 weeks. Then the biochemical and pathological indexes were determined and analyzed by factorial design ANOVA. Results After single intragastric administration of rifampicin, the serum bilirubin levels gradually increased, but no other indicators were affected in mice. Continuous intragastric administration of RIF 300 mg · kg-1, INH 150 mg · kg-1 or RIF 300 mg · kg-1 +INH 150 mg · kg-1 for 1 ∼3 weeks could significantly increase the liver index of mice. RIF alone or combined with isoniazid could significantly increase the level of ALT, AST and TBIL, resulting in vacuolar lesions in liver tissues in mice. The analysis of variance demonstrated that the combined use of RIF and INH for two weeks or three weeks showed significant antagonism in liver index and the level of ALT, and marked antagonism in TBIL at three weeks. The pathological results were basically consistent with the biochemical indicators. Conclusions RIF is the leading cause of liver injury in mice, and its hepatotoxicity is related to cholestasis. INH has a significant antagonistic effect on liver toxicity of RIF when they are combined, and the deep action mechanisms remains to be further explored.

Effect of sciatic nerve pulsed radiofrequency glial activation levels in spinal dorsal horn in chronic constriction injury rat models and its analgesia effect / 吉林大学学报(医学版)

Objective: To observe the effect of sea tic nerve pulsed radiofrequency (PRF) on the activation levels of microglia cells and astrocytes in spinal dorsal horn in the chronic constriction injury (CCD rat models, and to explore the relationships between the analgesic mechanism of sciatic nerve PRF and the activation levels of microglia cells and astrocytes in the spinal dorsal horn- Methods: Forty male SD rats were randomly divided into CCI shamoperation+RPF sham group (SS group). CC1 sham-operation + RPF group (SP group). CCI+RPF sham group (CS group). CCI+ RPF group (CP group) ( n 10). PRF was applied on the sciatic nerve on the 4th day after CC1 operation for 120 s, with a maximum temperature of 42"C. The mechanical withdrawal thresholds (MWT) and thermal paw withdrawal latencies 0. 05). Conclusion- PRF on sciatic nerve can relieve the neuropathic pain (NT) of the CCI rat models: PRF on saatic nerve can inhibit activation of the microglia cells in the spinal dorsal horn. The analgeac mechanism of PRF on sciatic nerve may be associated with the inhibition of the activation of microglia cells in the spinal dorsal horn.

Valproic acid inhibits the expressions of MMP-9 and AQP-4 proteins in the brain tissue of rats with traumatic brain injury / 医学研究生学报

Objective Few studies are reported on the protective effect of valproic acid (VPA) against traumatic brain injury (TBI) by down-regulating the protein expressions of matrix metalloproteinase-9 (MMP-9) and aquaporin-4 (AQP-4) in the brain tissue. This study aimed to investigate the neuroprotective effects of different doses of VPA against TBI in experimental rats. Methods We randomly divided 100 adult male rats into five groups of equal number, sham operation, TBI model, and low- (30 mg), medium- (150 mg) and high-dose (300 mg) VPA treatment. At 1, 3, 7 and 14 days after modeling by controlled cortex impact, we obtained the modified Neurological Severity Scores (mNSS), measured the VPA concentration in the venous blood, and then killed the rats and harvested the brain tissue for determination of the water content using the dry-wet method and the expressions of MMP-9 and AQP-4 by Western blot and immunohistochemistry. Results At 1, 3, 7 and 14 days after modeling, the mNSSs in the high-dose VPA group were 4.6 ± 1.3, 3.8 ± 1.3, 3.0 ± 0.7 and 1.8 ± 0.8, respectively, significantly lower than 8.4 ± 0.9, 7.0 ± 0.7, 5.8 ± 1.0 and 4.5 ± 1.3 in the TBI group (P < 0.05), decreasing in a time-dependent manner, with statistically significant difference between any two dose groups (P < 0.05). At 1, 3 and 7 days, the water contents in the brain tissue were (76.2 ± 0.7)%, (76.9 ± 1.7)% and (73.9 ± 1.3)% in the high-dose VPA group, significantly lower than (79.6 ± 0.8)%, (82.6 ± 0.8)% and (78.6 ± 0.7)% in the TBI group (P < 0.05), also decreasing in a time-dependent manner, with statistically significant difference between any two dose groups (P < 0.05). At 1 and 3 days, the expressions of MMP-9 and AQP-4 in the brain tissue were markedly down-regulated in the VPA groups in a dose-dependent manner as compared with those in the TBI group (P < 0.05), with statistically significant difference between any two dose groups (P < 0.05), and meanwhile immunohistochemistry showed large numbers of cells with positive expressions of MMP-9 and AQP-4, which were reduced with the increased dose of VPA. Conclusion VPA has a neuroprotective effect against TBI in rats by inhibiting the expressions of MMP-9 and AQP-4 proteins in the brain tissue and alleviating brain edema. Within the range of the doses studied, higher-dose VPA produces a better effect.

Effects of Dihydroquercetin on Hemorheology and Other Relevant Indexes in Local Cerebral Ischemic Injury Model Rats / 中国药房

OBJECTIVE： To observe the effects of dihydroquercetin （DHQ） on hemorheology and other relevant related indexes in local cerebral ischemic injury model rats. METHODS： SD rats were randomly divided into sham operation group， model group， nimodipine group （positive control， 20 mg/kg） and DHQ low-dose， medium-dose and high-dose groups （15， 30， 60 mg/kg）， with 10 rats in each group. Administration groups were given relevant medicine intragastrically， sham operation group and model group were given constant volume of 0.4％ Sodium carboxymethyl cellulose solution， once a day， for consecutive 14 d. After last administration， local cerebral ischemic injury model was induced by bilateral common carotid artery ligation in other groups except for sham operation group. After 24 h of cerebral ischemia， histopathological changes of brain tissue in rats of each group were observed； the levels of hemorheology indexes [whole blood viscosity （low， medium and high shear）， whole blood reduced viscosity （low， medium and high shear）， plasma viscosity]， erythrocyte parameters （hematocrit， EAI， DI， IR）， coagulation function indexes （APTT， PT， TT， FIB） were detected. RESULTS： Compared with sham operation group， the cells in the brain tissue of model group were loose， the gap was obvious， and the neurons around the ischemic area were damaged obviously； the levels of whole blood viscosity， whole blood reduced viscosity， plasma viscosity， hematocrit， EAI， IR and FIB were increased significantly， while the levels of DI， APTT， PT and TT were decreased or shortened significantly （P＜0.05 or P＜0.01）. Compared with model group， above symptoms of administration groups were improved to different extents， whole blood viscosity， plasma viscosity， EAI and IR of nimodipine group， whole blood viscosity and hematocrit of DHQ high-dose group， plasma viscosity and EAI of DHQ groups， and IR of DHQ medium-dose and high-dose groups were decreased significantly； DI， APTT， PT and TT of nimodipine group， DI， APTT and TT of DHQ groups and PT of DHQ high-dose group were increased or prolonged significantly （P＜0.05 or P＜0.01）. There was no statistical significance in other indexes among those groups （P＞0.05）. CONCLUSIONS： DHQ can protect against local cerebral ischemic injury model rats， the mechanism of which may be associated with improving hemorheology indexes and coagulation function disorder.

A study on goat models for training combat medics' key first aid skills on battle field / 中国中西医结合急救杂志

Objective To train the combat medics to learn key battle field operative technologies such as tracheotomy, thoracic close drainage, control of massive hemorrhage, etc by practicing the above skills on goats' traumatic models. Methods From 2012 to 2014 for consecutive 3 years, a group army trained the combat medics to learn how to treat war trauma first aid skills every year. 30 combat medics were randomly selected from 136 combat medics who had received goat model training to be tested. Goat model preparation methods: 35 healthy adult goats were anaesthetized with ketamine, then the suffocation models were reproduced by wadding the goat mouths and noses;pneumothorax models were replicated by cutting goat chest cavities; massive hemorrhagic models were produced by cutting goat femoral arteries by scissors. 136 combat medics were trained to perform tracheotomy, tube thoracostomy or thoracic close drainage, and hemorrhagic control by above models. The differences in mastering these skills before training, immediately after training, and one year after training were recorded and compared, and the factors influencing the combat medics training grade were analyzed. Results At the end of the training, the 30 combat medics' successful rates of performing cricothyrotomy or tracheotomy, tube thoracostomy, hemorrhage control were significantly higher than those before the training [respectively was 63.3% (19/30) vs. 10.0% (3/30), 66.7% (20/30) vs. 13.3% (4/30), 86.7% (26/30) vs. 53.3% (16/30), all P < 0.05]. After 1 year of training, the success rates of tracheotomy and thoracic close drainage were 33.3% (9/27) and 37.0% (10/27) respectively, which were significantly lower than those immediately after the end of training; the success rate of hemostasis after femoral artery rupture was 70.4% (19/27), which was lower than that at the end of training, but the difference was not statistically significant (P > 0.05). The education level had effect on the combat medics' performance after training. The success rate of combat medics with higher or above higher education was significantly higher than that of them with high school and below [88.9% (24/27) vs. 65.1% (41/63), P < 0.05]. Cross-sectional survey result showed that in 107 combat medics simultaneously received multimedia teaching, high analogue simulation human model teaching and animal model teaching, 85 combat medics (79.4%) chose the goat models as the first option for training. Conclusion By performing battle field key first aid techniques on goat trauma models, the combat medics' skills can be obviously elevated, they approve this animal model training as the first option, but repetition of the training is necessary to maintain the skills long lasting.

A novel zebrafish vascular injury model for asseessing drug efficancy of Yangxue Qingnao granules / 中国药理学与毒理学杂志

OBJECTIVE To assess the efficancy of Yangxue Qingnao granules on simvastatin-induced vascular injury model in zebrafish. METHODS Since statins can inhibit vascular development in zebrafish,in this study,we developed a novel animal model using 1 to 3 day post-fertilization larval zebrafish by optimizing the doses and duration of simvastatin exposure.Five pro-angiogenic drugs with a variety of mechanisms were tested to validate the newly developed zebrafish model. Zebrafish was treated with different concentration of Yangxue Qingnao granules( 62.5,125 and 250 μg·mL-1)for 2 d then tested for the area of subintestinal vein vessels. RESULTS Vascular regeneration promoting effect of five pro-angiogenic drugs (calycosin, astragaloside, chlorogenic acid, ferulic acid and Panax Notoginseng Saponins)were 8-48%,24-51%,35-58%,28-75% and 37-69%,respectively.In 62.5,125 and 250 μg·mL-1Yangxue Qingnao granules group,vascular regeneration promoting effect were 21% (P>0.05), 84%(P<0.01) and 53%(P<0.01). CONCLUSION Our results demonstrate that the zebrafish vascular injury model validated in this study could be used for in vivo angiogenesis studies and drug screening and for assessing pro-angiogenic drugs with different mechanisms.Yangxue Qingnao granules could promote the vascular regeneration in zebrafish.

Experimental study on mouse acute toxicity and analgesic pharmacodynamics of new compound L11 / 中国药理学通报

Aim To observe the effect of new com-pound L11 on the affinity and function of sigma-1 re-ceptor,as well as the mouse acute toxicity and analge-sic effect, so as to provide the experimental basis for its pharmacodynamics and preliminary toxicity evalua-tion. Methods Using binding and function test of sig-ma-1 receptor in vitro, the acute toxicity and formalin model test of mice,as well as the rat chronic constric-tion injury(CCI) model test in vivo,the effects of L11 on the inhibitory rate and function of sigma-1 receptor, LD50,lifting/licking time of mice and mechanical pain threshold of rats were respectively measured to evaluate the analgesic effect and mechanism of L11. Results The inhibitory rate and Kiof L11 on the sigma-1 recep-tor were 103.07% and 4.81 nmol·L-1,respectively. The Kivalue was 8.10 nmol·L-1while adding pheny-toin (sigma-1 receptor allosteric modulator). The in-tragastric administration of L11 in mice was 1 680.03 mg·kg-1LD50,and the 95% confidence interval was (1 559.35 ~1 819.40) mg·kg-1. Compared with model group, the II phase lifting/licking time of mice was significantly reduced and the mechanical pain threshold of rat obviously increased by L11. Conclu-sions The new compound L11 has high affinity to sig-ma-1 receptor, which belongs to the antagonist of sig-ma-1 receptor;L11 is less toxic to intragastric adminis-tration and has obvious analgesic effect on the formalin model of mice and CCI model of rats, which may be relative with the sigma-1 receptor antagonism.

Anti-oxidative stress effects of curcumin on rat models of traumatic brain injury / 中国比较医学杂志

Objective To investigate the changes of content or activity of Nrf2 and anti-oxidative stress-related factors in rat models of traumatic brain injury,and explore the mechanisms of protective effect of curcumin on brain damage and oxidative stress in rats. Methods Twenty healthy SPF male SD rats were divided into 4 groups: the control group, brain injury model group(TBI group),brain injury and solvent-treated group(TBI+S group),brain injury and curcumin-treated group(TBI+C group),5 rats ineach group. The control group received only saline and anesthesia. The TBI,TBI+S and TBI+C groups were given free falling body brain injury modeling device to establish the models and then received curcumin(5 mg/kg), an equal amount of DMSO solvent(0.05%)and an equal amount of physiological saline, respectively. The rats were sacrificed at the next day and the RNA and proteins of brain tissues were extracted. qRT-PCR and Western blot were used to detect the mRNA and protein expression of Nrf2. Chemocolorimetry was used to detect the content or activity of MDA, GSH, CAT and SOD in the brain tissues of rats. ELISA was used to detect the contents of iNOS and HO-1. Results Compared with the control group,the mRNA and protein expressions of Nrf2, the content of MDA,the activity of HO-1 and iNOS were significantly increased,the content of GSH, the activity of SOD and CAT were significantly decreased in the TBI group and TBI+S group,with a significant difference(P< 0.05). Compared with the TBI and TBI+S groups,the mRNA and protein expressions of Nrf2,the content of MDA,the activities of HO-1 and iNOS were significantly decreased,the content of GSH,the activity of SOD and CAT were significantly increased in the TBI+C group,showing a sigfnificant difference(P< 0.05),but there were no significant differences between the TBI group and TBI+S group(P< 0.05). Conclusions Curcumin has an anti-oxidative stress effect on rats with brain injury. It can reduce the expression of Nrf2,change the anti-oxidant stress-related indicators,therefore to protect the TBI-impaired brain tissues.

Study on pharmacokinetics and pharmacodynamics of ginkgolides in rat model with cerebral ischemia injury / 中草药

Objective To study the metabolic process of ginkgolides in rats with cerebral ischemic injury based on pharmacokinetic- pharmacodynamics (PK-PD) binding model. Methods The middle cerebral artery occlusion (MCAO) model was established by the suture method. After reperfusion, rats were randomly assigned to nasal administration, ig administration, and iv administration group.The orbital blood was taken at different time points of 0.25, 0.33, 0.5, 0.75, 1.0, 1.25, 1.5, 2.0, 4.0, 6.0, 9.0, and 12.0 h after the administration of the ginkgolides solution. The drug-time curve of ginkgolide B in plasma were drawn according to the concentration measured by LC-MS. The time-effect curve of superoxide dismutase (SOD) and malondialdehyde (MDA) were drawn based on the value measured by the kit. The pharmacokinetic parameters were calculated by DAS 2.0 software to fit the PK-PD binding model. Results The t1/2 of ginkgolide B of the rats in the administration group was smaller than that in the MCAO model group. The area under the curve (AUC) of nasal administration was significantly higher than intragastric administration and intravenous administration. Conclusion Ginkgolide B has a good protective and mitigating effect on ischemic stroke. The pharmacokinetics of nasal administration is better than iv and ig administration, which can provide reference for the development of nasal administration of ginkgolide B.

Quality evaluation of Gualou Guizhi decoction based on chemical compositions and biological effects / 中国中药杂志

The paper was aimed to establish a quality evaluation model for Gualou Guizhi decoction based on the chemical compositions and biological effects. Ultra high performance liquid chromatograph-mass spectrometer was used to analyze and determine 24 kinds of chemical compositions in Gualou Guizhi decoction, and then, biological activity effect was quantitatively assessed in a zebrafish neuronal injury model which was induced by mycophenolate mofetil (MMF). As a result, the established method for quality evaluation of Gualou Guizhi decoction based on the chemical compositions and biological effects is feasible, stable and reliable, which can provide reference for quality control of compound Chinese medicines.

Validation Study on Efficacy Difference of Green Forsythiae Fructus and Ripe Forsythiae Fructus in the Classic Formula / 中国中医药信息杂志

Objective To compare the efficacy difference of Green Forsythiae Fructus and Ripe Forsythiae Fructusby using Lianqiao powder and Yinqiao powder in the classic formula; To provide experimental evidence for the guidance of one for dual-use of the Forsythiae Fructus. Methods The guinea pig sore model was made with Staphylococcus aureus. 40 guinea pigs were randomly divided into blank group, model group, Lianqiao powder Green Forsythiae Fructus group and Lianqiao powder Ripe Forsythiae Fructus group. The blank group and the model group were fed with normal saline, while Lianqiao powder Green Forsythiae Fructus group and Lianqiao powder Ripe Forsythiae Fructus group were treated with 1.2 g raw medicine/kg liquid Lianqiao powder Green Forsythiae Fructus and Lianqiao powder Ripe Forsythiae Fructus for 7 d. The symptom score, blood and pathological changes of guinea pig soreness were observed. The model of acute lung injury was induced by 10% LPS aerosol inhalation. 40 rats were randomly divided into blank group, model group, Yinqiao powder Green Forsythiae Fructus group and Yinqiao powder Ripe Forsythiae Fructus group. The blank group and the model group were fed with normal saline, while Yinqiao powder Green Forsythiae Fructus group and Yinqiao powder Ripe Forsythiae Fructus group were treated with 4 g raw medicine/kg liquid Yinqiao powder Green Forsythiae Fructus and Yinqiao powder Ripe Forsythiae Fructus for 10 d. The levels of IL-6, TNF-α and IL-1β in the lung lavage fluid were detected. Results The effect of Lianqiao powder Green Forsythiae Fructus on the wound healing of guinea pig sore wound was faster than that of Lianqiao powder Ripe Forsythiae Fructus, but there was no significant difference between Lianqiao powder Green Forsythiae Fructus group and Lianqiao powder Ripe Forsythiae Fructus group in inhibiting the secretion and pathological changes of guinea pig sore wound. The levels of IL-6, TNF-α and IL-1β in Yinqiao powder Green Forsythiae Fructus group was lower than that in Yinqiao powder Green Forsythiae Fructus group, without statistical significance. Conclusion It is verified that there is efficacy differences between Green Forsythiae Fructus and Ripe Forsythiae Fructus in the different Chinese herbal compound.

Abirritation of geraniol on neuropathic pain model rat and possible mechanism / 中国药理学通报

Aim To study the analgesic effect of geraniol on neuropathic pain and to explore the possible mechanism.Method A neuropathic pain rat model of Spared Nerve Injury(SNI) was established to measure changes in the threshold of paw withdrawal before and after i.p.administration of geraniol.Patch clamp whole-cell recording was performed to measure activity of sodium channels using ipsilateral L3/L4/L5 dorsal root ganglion(DRG) cells isolated from the SNI rats.In addition, HEK 293 cells expressing hNav1.7 and hTRPA1 channels were used for measuring the changes in channel activities with or without geraniol by whole-cell patch clamp.Results Geraniol had a fast analgesic effect on hypersensitivity of mechanical pain in the SNI model.It significantly inhibited sodium channels on DRGs isolated from SNI rats and hNav1.7 but not hTRPA1 channels expressed by HEK293 cells.However, high concentrations of geraniol facilitated the activation of HTRPA1 channel stimulated by AITC.Conclusion Geraniol may abirritate hypersensitivity of mechanical pain in the SNI model by specifically inhibiting Nav1.7 channel activity on the DRG cells.

Effect of Five Kinds of CYP450 Enzymes in Different Acute Liver Injury Rat Models / 中国药学杂志

OBJECTIVE: To compare the effect on CYP450 isoenzyme in rats with acute liver injury induced by different chemicals. METHODS: Acute liver injury model of rats induced by tetrachloromethane(CCl4), D-aminogalactose(D-GalN)/lipopolysaccharide(LPS), α-naphthyl isothiocyanate(ANIT) respectively whereas the normal Wistar rats were used as controls. After the tail vein injection with Cocktail probe solutions prepared with five CYP450s probe substrates (phenacetin-CYP1A2, omeprazole-CYP2C9, tolbutamide-CYP2C19, dextromethorphan-CYP2D6, midazolam-CYP3A4), the blood samples were collected from the fundus venous plexus of rat at different time point, the blood drug concentration of the five probe substrates were determined by LC-MS/MS, and the pharmacokinetic parameters were calculated by PK Solutions 2™. Compared with normal rats, the changes of the probe drug pharmacokinetics in different rat models were used as the basis for the evaluation of the metabolic activity of CYP450 isoenzyme. RESULTS: Compared with normal rats, the activities of CYP1A2, CYP2C9, CYP2C19, CYP2D6 of CCl4 group rats were significantly inhibited, and the activities of CYP3A4 was slightly inhibited; the activities of CYP2C9, CYP2D6 and CYP3A4 of D-GalN/LPS group rats were significantly induced, and the activity of CYP2D6 and CYP3A4 was slightly induced, and the activity of CYP1A2 was not significantly affected, but the activity of CYP2C19 was significantly inhibited; the activities of CYP2C9, CYP2C19 and CYP3A4 of ANIT group rats were significantly induced, the activity of CYP3A4 were slightly induced, and the activity of CYP2D6 was not significantly affected, but the activity of CYP1A2 was significantly inhibited. CONCLUSION: There are significant differences in the activities of CYP450 isoenzyme in the rat model of acute liver injury induced by different chemicals.

Hypertrophy of rat myocardial cells injury model:a short discuss / 国际药学研究杂志

Myocardiel hypertrophy,a common cardiovascular and cerebrovascular disease,has attracted more and more atten?tion worldwide in recent years. Investigation of its pathogenesis is important for its prevention and treatment. The choice and establish?ment of experimental models play a vital role in these investigations. A proper experimental model can not only reduce costs and labor , but also reflect the nature of the experimental phenomena,which helps to achieve adequate experimental purposes. In the review,we introduce a brief discussion of on currently used primary cellular model of myocardial hypertrophy to provide reference for ventricular remodeling diseases researches.

The Effect of DA-6034 on Intestinal Permeability in an Indomethacin-Induced Small Intestinal Injury Model

BACKGROUND/AIMS: DA-6034 has anti-inflammatory activities and exhibits cytoprotective effects in acute gastric injury models. However, explanations for the protective effects of DA-6034 on intestinal permeability are limited. This study sought to investigate the effect of DA-6034 on intestinal permeability in an indomethacin-induced small intestinal injury model and its protective effect against small intestinal injury. METHODS: Rats in the treatment group received DA-6034 from days 0 to 2 and indomethacin from days 1 to 2. Rats in the control group received indomethacin from days 1 to 2. On the fourth day, the small intestines were examined to compare the severity of inflammation. Intestinal permeability was evaluated by using fluorescein isothiocyanate-labeled dextran. Western blotting was performed to confirm the association between DA-6034 and the extracellular signal-regulated kinase (ERK) pathway. RESULTS: The inflammation scores in the treatment group were lower than those in the control group, but the difference was statistically insignificant. Hemorrhagic lesions in the treatment group were broader than those in the control group, but the difference was statistically insignificant. Intestinal permeability was lower in the treatment group than in the control group. DA-6034 enhanced extracellular signal-regulated kinase expression, and intestinal permeability was negatively correlated with ERK expression. CONCLUSIONS: DA-6034 may decrease intestinal permeability in an indomethacin-induced intestinal injury model via the ERK pathway.

Enhanced Expression of TREK-1 Is Related with Chronic Constriction Injury of Neuropathic Pain Mouse Model in Dorsal Root Ganglion

Neuropathic pain is a complex state showing increased pain response with dysfunctional inhibitory neurotransmission. The TREK family, one of the two pore domain K+ (K2P) channel subgroups were focused among various mechanisms of neuropathic pain. These channels influence neuronal excitability and are thought to be related in mechano/thermosensation. However, only a little is known about the expression and role of TREK-1 and TREK-2, in neuropathic pain. It is performed to know whether TREK-1 and/or 2 are positively related in dorsal root ganglion (DRG) of a mouse neuropathic pain model, the chronic constriction injury (CCI) model. Following this purpose, Reverse Transcription Polymerase Chain Reaction (RT-PCR) and western blot analyses were performed using mouse DRG of CCI model and compared to the sham surgery group. Immunofluorescence staining of isolectin-B4 (IB4) and TREK were performed. Electrophysiological recordings of single channel currents were analyzed to obtain the information about the channel. Interactions with known TREK activators were tested to confirm the expression. While both TREK-1 and TREK-2 mRNA were significantly overexpressed in DRG of CCI mice, only TREK-1 showed significant increase (~9 fold) in western blot analysis. The TREK-1-like channel recorded in DRG neurons of the CCI mouse showed similar current-voltage relationship and conductance to TREK-1. It was easily activated by low pH solution (pH 6.3), negative pressure, and riluzole. Immunofluorescence images showed the expression of TREK-1 was stronger compared to TREK-2 on IB4 positive neurons. These results suggest that modulation of the TREK-1 channel may have beneficial analgesic effects in neuropathic pain patients.