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1.
Article in Chinese | WPRIM | ID: wpr-928734

ABSTRACT

OBJECTIVE@#To investigate the mechanism of the in vitro toxicity of doxycycline to myeloma cell line H929 and also the possible pathway involved its toxicity.@*METHODS@#Myeloma cell line H929 was treated with DOX, MEK inhibitor U0126 or RAS agonist ML-098, either alone or in combination. Then, the expression of p-MEK, caspase-3, caspase-9 and c-Jun in H929 were used to detected by Western blot; the cells proliferation and apoptosis were detected by CCK-8 assay and flow cytometry, respectively.@*RESULTS@#DOX significantly increased the levels of cleaved caspase-3 and caspase-9, and down-regulated the level of p-MEK in H929 (P<0.05). MEK antagonist U0126 significantly increased the levels of cleaved caspase-3 and caspase-9, and down-regulated the level of p-MEK (P<0.05). After Dox combined with ML-098 treatment of H929 cells, the apoptosis rate of H929 cells was lower than that of DOX alone treatment group(P<0.05). Compared with DOX alone treatment group, the expressions of p-MEK and p-ERK1/2 in DOX+ML-098 combined treatment group were increased, and the levels of cleaved caspase-3,9 in H929 cells were decreased (P<0.05). The levels of c-Jun mRNA and protein increased in H929 when treated by DOX alone (P<0.05).@*CONCLUSION@#DOX can induce apoptosis of H929 via intrinsic apoptosis pathway, and MEK/ERK pathway and c-Jun possibly play a role in this process.


Subject(s)
Apoptosis , Caspase 3 , Caspase 9/pharmacology , Cell Line, Tumor , Cell Proliferation , Doxycycline/pharmacology , Humans , Mitogen-Activated Protein Kinase Kinases/pharmacology , Multiple Myeloma
2.
Article in Chinese | WPRIM | ID: wpr-927895

ABSTRACT

Objective: To investigate the repair effect and JNK/NF-κB,SOX9 mechanisms of vibration exercise with different frequencies on articular cartilage in rats with early knee osteoarthritis. Methods: Forty-eight adult male SD rats were randomly divided into six groups(n=8):model control group(MC),high frequency vibration group 1 (GP1,60 Hz),high frequency vibration 2 group (GP2,40 Hz),medium frequency vibration group (ZP,20 Hz),minor frequency group(DP,10 Hz)and normal control group(NC). Except for NC group,the rats in each group were made into early knee osteoarthritis model after six weeks of knee joint cavity injection of papain solution and 2% mixture l-cysteine on the 1st,4 th and 7th day. Each exercise group was subjected vibration to 40 minutes a day with amplitude of 2~5 mm and 5 days a week. Four weeks later, the articular cartilage of the lateral femoral condyle of the both back leg knee joints were detected by HE staining,serine O staining and Mankin scores for morphological observation. The expression levels of JNK,NF-κB p65 and Sox9 mRNA in articular cartilage of the medial femoral condyle were detected by RT-qPCR,and the protein expressions of JNK,NF-κB p65 and Sox9 were detected by Western blot. Results: Compared with the NC group,the Mankin score in other groups was significantly higher (P<0.01). Compared with the MC group,the Mankin score of each vibration group was significantly lower(P<0.05),the mRNA and protein expressions of JNK and NF-κB p65 in each vibration training group were significantly lower (P<0.01),the expressions of Sox9 mRNA and protein in vibration training group were increased significantly (P<0.01). Compared with the higher frequency group,the Mankin score,the mRNA and protein expressions of JNK and NF-κB p65 of lower frequency group were significantly lower (P<0.05 or P<0.01). But the expressions of Sox9 mRNA and protein were significantly higher (P< 0.05 or P<0.01). Conclusion: Vibration exercise of different frequencies may present varying degrees of cartilage repair impact in rats with early knee osteoarthritis,and the cartilage repair by low-frequency vibration training is better than that by high-frequency vibration. This can be one of the mechanisms on controlling collagen synthesis by down-regulating JNK/NF-κB expression and increasing SOX9 activity of OA articular cartilage.


Subject(s)
Animals , Cartilage, Articular/metabolism , MAP Kinase Kinase 4 , Male , NF-kappa B/metabolism , Osteoarthritis, Knee/therapy , RNA, Messenger/metabolism , Rats , Rats, Sprague-Dawley , SOX9 Transcription Factor , Vibration
3.
Chinese Journal of Endemiology ; (12): 100-104, 2022.
Article in Chinese | WPRIM | ID: wpr-931501

ABSTRACT

Objective:To investigate the expression and significance of endoplasmic reticulum stress apoptosis pathway related proteins in renal cortex of rats with chronic fluorosis.Methods:Twenty four healthy SD rats were divided into 4 groups (6 rats/group, half male and half female) according to their body mass (100 - 120 g) by random number table method, rats in control group drank tap water (fluoride content < 0.5 mg/L), and in low, medium and high fluoride groups drank tap water with fluoride content (sodium fluoride) of 10, 50 and 100 mg/L, respectively. After 180 days of feeding, dental fluorosis was examined, 24-hour urine sample was collected and the content of fluoride in urine was detected by fluoride ion selective electrode method. Renal tissue was taken after anesthesia, and the pathological changes of renal cortex were observed by hematoxylin-eosin (HE) staining. The expressions of endoplasmic reticulum stress apoptosis pathway related proteins [inositol-requiring enzyme 1α (IRE1α), apoptosis signal-regulating kinase 1 (ASK1), C-Jun N-terminal kinase (JNK) and phosphorylated JNK (P-JNK)] were determined by immunohistochemical staining in rat renal cortex.Results:No dental fluorosis was found in control group. The incidence of dental fluorosis in low, medium and high fluoride groups were 2/6, 5/6 and 6/6, respectively. Compared with control group [(5.707 ± 1.190) mg/L], the urinary fluoride in low, medium and high fluoride groups [(17.028 ± 3.006), (34.378 ± 12.045), (94.759 ± 31.773) mg/L] was significantly higher ( P < 0.05), and the urinary fluoride in high fluoride group was higher than that in low and medium fluoride groups ( P < 0.05). HE staining showed that, compared with control group, the cell volume of renal tubules and glomeruli in medium and high fluoride groups increased, the cells arranged closely, and the eosinophilia of the cytoplasm increased. The immunohistochemical staining results showed that there was no significant difference in the expression of JNK protein in rat renal cortex between control group and low, medium and high fluoride groups ( F = 0.07, P > 0.05). The expressions of IRE1α, ASK1 and P-JNK proteins in rat renal cortex in high fluoride group were higher than those of control, low and medium fluoride groups ( P < 0.05), and the expressions of IRE1α and ASK1 proteins in medium fluoride group were significantly higher than those in control and low fluoride groups ( P < 0.05). Conclusion:Long-term excessive fluoride intake can lead to renal cortex injury in rats, and the mechanism of injury may be related to the activation of IRE1α-ASK1-JNK endoplasmic reticulum stress apoptosis pathway.

4.
Acta Pharmaceutica Sinica B ; (6): 2129-2149, 2022.
Article in English | WPRIM | ID: wpr-929399

ABSTRACT

Cardiometabolic disease (CMD), characterized with metabolic disorder triggered cardiovascular events, is a leading cause of death and disability. Metabolic disorders trigger chronic low-grade inflammation, and actually, a new concept of metaflammation has been proposed to define the state of metabolism connected with immunological adaptations. Amongst the continuously increased list of systemic metabolites in regulation of immune system, bile acids (BAs) represent a distinct class of metabolites implicated in the whole process of CMD development because of its multifaceted roles in shaping systemic immunometabolism. BAs can directly modulate the immune system by either boosting or inhibiting inflammatory responses via diverse mechanisms. Moreover, BAs are key determinants in maintaining the dynamic communication between the host and microbiota. Importantly, BAs via targeting Farnesoid X receptor (FXR) and diverse other nuclear receptors play key roles in regulating metabolic homeostasis of lipids, glucose, and amino acids. Moreover, BAs axis per se is susceptible to inflammatory and metabolic intervention, and thereby BAs axis may constitute a reciprocal regulatory loop in metaflammation. We thus propose that BAs axis represents a core coordinator in integrating systemic immunometabolism implicated in the process of CMD. We provide an updated summary and an intensive discussion about how BAs shape both the innate and adaptive immune system, and how BAs axis function as a core coordinator in integrating metabolic disorder to chronic inflammation in conditions of CMD.

5.
Article in Chinese | WPRIM | ID: wpr-940754

ABSTRACT

ObjectiveTo investigate the mechanism of Huangqi Guizhi Wuwutang (HQGZWWT) in the treatment of diabetic peripheral neuropathy (DPN) in MKR mice via regulating endoplasmic reticulum (ER) stress. MethodThirty-two 8-week-old MKR mice (half were male and half were female) were fed with a high-fat diet for four weeks, and then 1% streptozotocin (STZ) was injected intraperitoneally for five days. After the blood glucose was stabilized, the mice were housed in the cage covered with ice bags for another one hour stimulation per day for four weeks. Mice with fasting blood glucose (FBG) value ≥11.1 mmol·L-1 were randomly divided into model group , Huangqi Guizhi Wuwutang in original dosage group (30 g·kg-1·d-1), Huangqi Guizhi Wuwutang in formula dosage group (6.25 g·kg-1·d-1), and positive drug group (mecobalamin tablets, 0.17 mg·kg-1·d-1). Another eight MKR mice of the same age were set as blank group and eight FVB mice were normal group. After four weeks of intragastric administration in each group, the change in FBG was tested, and hematoxylin and eosin (HE) staining and transmission electron microscope were used for observing the morphology of sciatic nerve tissue. In addition, the expression of c-Jun N-terminal kinase (JNK), phosphorylated c-Jun N-terminal kinase (p-JNK) and inositol requiring enzyme 1α (IRE1α) proteins was determined by immunohistochemical test and Western blot (WB). ResultCompared with the conditions in the normal group and blank group, the time of paw withdrawal, paw licking and tail flick in the model group was shortened (P<0.01), and the conduction velocity of sciatic nerve was decreased (P<0.01). Compared with the conditions in the model group, the behavioral and functional indicators were improved by HQGZWWT (P<0.05,P<0.01). The immunohistochemical test revealed the JNK expression was elevated in the model group compared with the conditions in the normal group and blank group (P<0.05), while that was lowered by HQGZWWT compared with the condition in the model group (P<0.05). However, there was no difference among the treatment groups. According to the WB, the expression of IRE1α and p-JNK in the model group was enhanced compared with the conditions in the normal group and blank group (P<0.05,P<0.01), while that was decreased by HQGZWWT compared with the condition in the model group (P<0.05,P<0.01). No difference was observed between the HQGZWWTO and HQGZWWTF groups. ConclusionHQGZWWT can improve the neurophysiological function and pathological damage of sciatic nerve, which may be related to its delaying the ER stress response of sciatic nerve.

6.
Article in Chinese | WPRIM | ID: wpr-940700

ABSTRACT

ObjectiveTo identify the protective effect and possible mechanism of Gandou Fumu decoction (GDFMD) on liver fibrosis in mice with Wilson's disease. MethodA total of 50 homozygous TXJ mice were randomly divided into five groups, with 10 mice in each group. Ten wild-type mice were selected as a normal group. The GDFMD high, medium, and low-dose groups were given 13.92, 6.96, 3.48 g·kg-1 of GDFMD, respectively. The penicillamine group were given 0.1 g·kg-1 of penicillamine. The model group and the normal group were given the same volume of 0.9% sodium chloride solution once a day for 4 consecutive weeks. The enzyme-linked immunosorbent assay (ELISA) method was performed to detect serum superoxide dismutase (SOD) activity and malondialdehyde (MDA) content. Corresponding kits were used to detect the mitochondrial adenine triphosphate (ATP) content and Na+-K+-ATPase activity in liver tissues. Hematoxylin-eosin (HE) and Masson staining were used to observe the pathological morphology of liver tissue, and transmission electron microscope was used to observe ultrastructural changes of liver tissues in mice. Western blot was used to detect the c-Jun N-terminal kinase, the phosphorylated protein, and the expressions of Caspase-3, B cell lymphoma-2 (Bcl-2), and Bcl-2 associated X protein (Bax) in c-Jun N-terminal kinase (JNK) signaling pathway. ResultCompared with the normal group, MDA content increased and SOD activity decreased in the model group (P<0.05). Compared with the model group, SOD activities in the GDFMD high-, medium-, and low-dose groups and the penicillamine group significantly increased (P<0.01), and MDA content significantly decreased (P<0.05, P<0.01). Compared with the normal group, ATP content and Na+-K+-ATPase activity significantly decrease in the model group (P<0.05). Compared with the model group, ATP content and Na+-K+-ATPase activity in the GDFMD medium and high-dose groups and the penicillamine group significantly increased (P<0.05, P<0.01). The results of the pathological morphology of liver tissue showed that a large number of liver cells degeneration and necrosis, inflammatory cell infiltration, unclear liver lobule structure, and collagen fiber deposition were observed in the model group. Transmission electron microscopy showed that the number of mitochondria in liver tissues significantly reduced, the mitochondria were locally damaged, and the cristae of mitochondria were broken even disappear in the model group. The pathological morphology of liver tissue and mitochondrial structure recovered to varying degrees after medicinal intervention. The results of Western blot suggested that, compared with the normal group, the expression levels of phosphorylation-JNK (p-JNK), p-JNK/JNK, Caspase-3, and Bax in the liver tissues were up-regulated, while the expression of Bcl-2 was down-regulated in the model group (P<0.05). The expression levels of p-JNK, p-JNK/JNK, Caspase-3 and Bax were down-regulated and the expression of Bcl-2 was up-regulated in the GDFMD high and medium-dose groups and the penicillamine group (P<0.01). ConclusionGDFMD can alleviate oxidative stress damage and recover mitochondrial function of TXJ mice with liver fibrosis. The mechanism of GDFMD may be related to regulating the JNK signaling pathway and downstream factors and inhibiting cell apoptosis.

7.
Article in Chinese | WPRIM | ID: wpr-940453

ABSTRACT

ObjectiveTo explore the differences in the protective effects of five formulas for promoting blood circulation and removing blood stasis on the aortic endothelial cells of New Zealand rabbits with heart blood stasis syndrome. MethodEighty New Zealand rabbits were randomly divided into a normal group (n=10) and an experimental group (n=70). The heart blood stasis syndrome model was induced by starvation combined with a high-fat diet and adrenaline in the rabbits of the experimental group. Subsequently, the model rabbits were randomly divided into a model group, a Xuefu Zhuyutang group (3.55 g·kg-1·d-1), a Taohong Siwutang group (2.66 g·kg-1·d-1), a Danshenyin group (1.962 g·kg-1·d-1), a Huoluo Xiaolingdan group (2.80 g·kg-1·d-1), a Shixiaosan group (0.56 g·kg-1·d-1), and a c-Jun N-terminal kinase (JNK) inhibitor (SP600125, 5 μg·kg-1)group. The normal group and the model group received the same amount of distilled water. The rabbits in five Chinese medicine groups were treated correspondingly by gavage, and those in the SP600125 group were injected with 0.5 mL of SP600125-dimethyl sulfoxide diluent. After the treatment, the aorta was collected, and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was used to detect the apoptosis of aortic endothelial cells. The enzyme-linked immunosorbent assay (ELISA) was used to detect serum levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). Western blot was used to detect the protein expression of JNK, phosphorylated JNK (p-JNK), B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), cysteinyl aspartate-specific protease-9 (Caspase-9), and cysteinyl aspartate-specific protease-3 (Caspase-3) in aortic tissues. Real-time fluorescence-based quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA levels of JNK, Bcl-2, Bax, Caspase-9, and Caspase-3 in aortic tissues. ResultFive formulas could improve the apoptosis of aortic endothelial cells to varying degrees. To be specific, Xuefu Zhuyutang and Taohong Siwutang were optimal in efficacy, followed by Huoluo Xiaolingdan, Shixiaosan, and Danshenyin, and SP600125 was the worst (P<0.05, P<0.01). Five formulas could reduce the content of TNF-α and IL-6 (P<0.05, P<0.01), down-regulate the protein expression levels of JNK, p-JNK, Bax, Caspase-9, and Caspase-3 (P<0.05, P<0.01), decrease the mRNA expression levels of JNK, Bax, Caspase-9, and Caspase-3 (P<0.05, P<0.01), and up-regulate the protein and mRNA expression levels of Bcl-2 (P<0.05, P<0.01). ConclusionFive formulas can all reduce the apoptosis of aortic endothelial cells in New Zealand rabbits with heart blood stasis syndrome with different efficacies. It may be related to the different effects of five formulas on the JNK signaling pathway.

8.
Article in Chinese | WPRIM | ID: wpr-940348

ABSTRACT

ObjectiveTo investigate the effect of Jingangwan on the expression of osteoclast, c-Jun N-terminal kinase(JNK), p38 mitogen-activated protein kinase(p38 MAPK), and interleukin-1(IL-1) in the osteoporosis model rats, explore the mechanism of Jingangwan in the treatment of osteoporosis, and determine the optimal dosing concentration of Jingangwan. MethodFifty-six rats of SPF grade were randomized into a blank group,a sham operation group,a model group, model group,high-, medium-, and low-dose Jingangwan groups (0.72, 0.36, 0.18 g·kg-1·d-1, ig),and an estradiol valerate group (0.009 g·kg-1·d-1, ig), with eight rats in each group. The rats in the model group, the blank group, and the sham operation group received 3 mL of normal saline, respectively. Samples were collected 12 weeks after drug administration. The number of osteoclasts was observed by tartrate-resistant acid phosphatase (TRAP) staining. Serum levels of JNK, p38 MAPK, and IL-1 were detected by enzyme-linked immunosorbent assay (ELISA). The mRNA expression levels of p38 MAPK and JNK were detected by real-time quantitative polymerase chain reaction (Real-time PCR). ResultThe TRAP staining results showed that compared with the model group, the estradiol valerate group and the Jingangwan groups could inhibit the formation of osteoclasts to different degrees. As revealed by ELISA results, compared with the model group and the sham operation group, the model group showed increased serum levels of p38 MAPK, JNK, and IL-1 (P<0.01), while compared with the model group, all the groups with drug intervention showed decreased levels of p38 MAPK, JNK, and IL-1 (P<0.01). The serum levels of JNK and IL-1 in the high-dose Jingangwan group were lower than those in the estradiol valerate group (P<0.05). Real-time PCR results showed that compared with the blank group, the model group showed increased relative mRNA expression of p38 MAPK and JNK in the thighbone (P<0.01), while compared with the model group, all the groups with drug intervention showed decreased relative mRNA expression of p38 MAPK and JNK in the thighbone (P<0.01). ConclusionJingangwan can inhibit the formation of osteoblasts,reduce the diameter of the bone marrow cavity,improve bone quality,suppress the production of inflammatory factors,affect the metabolism of the MAPK signaling pathway,and blunt p38 MAPK and JNK activities to inhibit the differentiation and proliferation of osteoblasts and regulate bone metabolism, thereby preventing osteoporosis. Therefore,Jingangwan may be of application value in maintaining bone health and treating osteoporosis.

9.
Article in English | WPRIM | ID: wpr-939585

ABSTRACT

Objective@#Fine particulate matter (PM 2.5) is an air pollutant that has become of great concern in recent years. Numerous studies have found that PM 2.5 may contribute to lung cancer, but the pathogenesis has not yet been fully elucidated. In this study, we explored the roles of exosomes from bronchial epithelial cells in PM 2.5-promoted lung cancer metastasis.@*Methods@#Exosomes were isolated from cell supernatants. An animal model of lung metastasis (established by tail vein injection of A549-luc) and in vitro studies with lung cancer cell lines were used to investigate the effects of exosomes derived from PM 2.5-treated human bronchial epithelial cells (PHBE-exo).@*Results@#The animal experiments revealed that PHBE-exo-treated mice showed stronger luciferase activity and a larger relative metastatic region in the lungs, thus indicating that PHBE-exo promoted the metastatic potential of lung cancer. Additionally, PHBE-exo promoted the migration, invasion and epithelial-to-mesenchymal transition of lung cancer cells, in a manner mediated by activation of c-Jun N-terminal kinase.@*Conclusion@#These results implied that PM 2.5 may promote the development of lung cancer through exosomes derived from bronchial epithelial cells, thus providing a potential interventional target for lung cancer. These findings broadened our understanding of cancer-promoting mechanisms of environmental pollutants.


Subject(s)
Animals , Epithelial Cells/metabolism , Epithelial-Mesenchymal Transition , Exosomes/metabolism , Humans , Lung Neoplasms/metabolism , Mice , Particulate Matter/toxicity
10.
Acta Pharmaceutica Sinica B ; (6): 2726-2737, 2021.
Article in English | WPRIM | ID: wpr-888884

ABSTRACT

Integrins are transmembrane receptors that have been implicated in the biology of various human physiological and pathological processes. These molecules facilitate cell-extracellular matrix and cell-cell interactions, and they have been implicated in fibrosis, inflammation, thrombosis, and tumor metastasis. The role of integrins in tumor progression makes them promising targets for cancer treatment, and certain integrin antagonists, such as antibodies and synthetic peptides, have been effectively utilized in the clinic for cancer therapy. Here, we discuss the evidence and knowledge on the contribution of integrins to cancer biology. Furthermore, we summarize the clinical attempts targeting this family in anti-cancer therapy development.

11.
Acta Pharmaceutica Sinica B ; (6): 1708-1720, 2021.
Article in English | WPRIM | ID: wpr-888831

ABSTRACT

Stroke is considered a leading cause of mortality and neurological disability, which puts a huge burden on individuals and the community. To date, effective therapy for stroke has been limited by its complex pathological mechanisms. Autophagy refers to an intracellular degrading process with the involvement of lysosomes. Autophagy plays a critical role in maintaining the homeostasis and survival of cells by eliminating damaged or non-essential cellular constituents. Increasing evidence support that autophagy protects neuronal cells from ischemic injury. However, under certain circumstances, autophagy activation induces cell death and aggravates ischemic brain injury. Diverse naturally derived compounds have been found to modulate autophagy and exert neuroprotection against stroke. In the present work, we have reviewed recent advances in naturally derived compounds that regulate autophagy and discussed their potential application in stroke treatment.

12.
Article in English | WPRIM | ID: wpr-880713

ABSTRACT

This study aims to elucidate the antiproliferative mechanism of hydroxychavicol (HC). Its effects on cell cycle, apoptosis, and the expression of c-Jun N-terminal kinase (JNK) and P38 mitogen-activated protein kinase (MAPK) in HT-29 colon cancer cells were investigated. HC was isolated from

13.
Acta Pharmaceutica Sinica B ; (6): 1148-1157, 2021.
Article in English | WPRIM | ID: wpr-881190

ABSTRACT

As one of the most lethal diseases, pancreatic cancer shows a dismal overall prognosis and high resistance to most treatment modalities. Furthermore, pancreatic cancer escapes early detection during the curable period because early symptoms rarely emerge and specific markers for this disease have not been found. Although combinations of new drugs, multimodal therapies, and adjuvants prolong survival, most patients still relapse after surgery and eventually die. Consequently, the search for more effective treatments for pancreatic cancer is highly relevant and justified. As a newly re-discovered mediator of gasotransmission, hydrogen sulfide (H

14.
Acta Pharmaceutica Sinica B ; (6): 309-321, 2021.
Article in English | WPRIM | ID: wpr-881138

ABSTRACT

Cullin-RING ligases (CRLs) recognize and interact with substrates for ubiquitination and degradation, and can be targeted for disease treatment when the abnormal expression of substrates involves pathologic processes. Phosphorylation, either of substrates or receptors of CRLs, can alter their interaction. Phosphorylation-dependent ubiquitination and proteasome degradation influence various cellular processes and can contribute to the occurrence of various diseases, most often tumorigenesis. These processes have the potential to be used for tumor intervention through the regulation of the activities of related kinases, along with the regulation of the stability of specific oncoproteins and tumor suppressors. This review describes the mechanisms and biological functions of crosstalk between phosphorylation and ubiquitination, and most importantly its influence on tumorigenesis, to provide new directions and strategies for tumor therapy.

15.
Acta Pharmaceutica Sinica B ; (6): 3740-3755, 2021.
Article in English | WPRIM | ID: wpr-922437

ABSTRACT

Acetaminophen (APAP) is a widely used analgesic and antipyretic drug, which is safe at therapeutic doses but can cause severe liver injury and even liver failure after overdoses. The mouse model of APAP hepatotoxicity recapitulates closely the human pathophysiology. As a result, this clinically relevant model is frequently used to study mechanisms of drug-induced liver injury and even more so to test potential therapeutic interventions. However, the complexity of the model requires a thorough understanding of the pathophysiology to obtain valid results and mechanistic information that is translatable to the clinic. However, many studies using this model are flawed, which jeopardizes the scientific and clinical relevance. The purpose of this review is to provide a framework of the model where mechanistically sound and clinically relevant data can be obtained. The discussion provides insight into the injury mechanisms and how to study it including the critical roles of drug metabolism, mitochondrial dysfunction, necrotic cell death, autophagy and the sterile inflammatory response. In addition, the most frequently made mistakes when using this model are discussed. Thus, considering these recommendations when studying APAP hepatotoxicity will facilitate the discovery of more clinically relevant interventions.

16.
Article in Chinese | WPRIM | ID: wpr-877084

ABSTRACT

Objectives To study the effect of cadmium (Cd) on the proliferation and apoptosis of mouse spermatocyte (GC-2 spd) cells and explore the underlying molecular mechanism. Methods GC-2 spd cells were cultured with 0, 5, 10, 15, 20 and 30 μM CdCl2, respectively, for 24 hours. The cell viability and IC50 of Cd were estimated based on CCK-8 data. The apoptosis of GC-2 spd cells and cellular concentration of ROS were analyzed by flow cytometry after treatment of the cells with different concentrations of CdCl2 (0, 5, 10 μM) for 24 hours. The expression levels of JNK/c-Jun signaling pathway regulatory proteins, pro-apoptotic factor Bax and anti-apoptotic factor Bcl-2, were determined by Western blot. Results Cd inhibited the proliferation of GC-2 spd cells with IC50 value of 12.99 μM, 95% CI [11.95, 14.00]. Exposure to 5 and 10 μM CaCl2 resulted in increases in apoptosis and cellular ROS generation in a dose-dependent manner, which was statistically significant compared with the control (P 0.05), the phosphorylation level of JNK and c-Jun in Cd group was highly increased as compared to the control (P < 0.05). In addition, Cd exposure significantly increased the expression of Bax protein but decreased the expression Bcl-2 protein (P < 0.05). Conclusions Cadmium induces GC-2 spd cell apoptosis by increasing concentration of ROS and regulating the JNK/c-Jun signaling pathway.

17.
Article in Chinese | WPRIM | ID: wpr-905984

ABSTRACT

Objective:To explore<italic> </italic>the efficacy and mechanism of Danggui Shaoyaosan on rats of nonalcoholic fatty liver disease (NAFLD). Method:Sixrty SPF SD male rats were randomly divided into normal group, model group,essentiale (0.144 g·kg<sup>-1</sup>) and low, middle and high-dose of Danggui Shaoyaosan groups (2.44, 4.88, 9.76 g·kg<sup>-1</sup>). High fat diet were fed to bulid the NAFLD model, and each treatment group was given corresponding drugs at the same time. After 8 weeks, the serum and liver tissue were collected to detect the contents or activities of total cholesterol (TC), triglyceride (TG), alanine aminotransferase (ALT), aspartic acid aminotransferase (AST), tumor necrosis factor-<italic>α</italic> (TNF-<italic>α</italic>) and interleukin-10 (IL-10) in serum, the contents of TC, TG and free fatty acid (FFA) in liver tissue, Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) and Western blot were used to observe the gene and protein expressions of Toll-like receptor 4 (TLR4), myeloid different factory 88 (MyD88) and c-Jun n-terminal kinase (JNK) and the protein expression of phosphorylation JNK(p-JNK) in liver tissue. Hematoxylin-eosin (HE) staining and Oil red staining to observe the pathological morphological changes of liver. Result:Compared with control group, the contents or activities of TC, TG, ALT, AST and TNF-<italic>α</italic> in serum, the contents of TC, TG and FFA in liver and the gene and protein expressions of TLR4, MyD88, JNK, and the protein expression of p-JNK in liver tissue of model group were distinctly increased (<italic>P</italic><0.01), the content of IL-10 was significantly decreased (<italic>P</italic><0.01). Compared with model group, the contents or activities of TC, TG, ALT, AST and TNF-<italic>α</italic> in serum, the contents of TC, TG and FFA in liver and the mRNA and protein expressions of TLR4, MyD88 and JNK, and the protein expression of p-JNK in liver tissue of Danggui Shaoyaosan groups were significantly reduced (<italic>P</italic><0.05,<italic>P</italic><0.01), the content of IL-10 in serum of Danggui Shaoyaosan groups was distinctly increased(<italic>P</italic><0.05,<italic>P</italic><0.01), HE staining and Oil red staining show that the degree of liver steatosis was alleviated obviously by Danggui Shaoyaosan. Conclusion:Danggui Shaoyaosan has a better treatment on NAFLD by inhibiting TLR4/MyD88/JNK pathway and alleviating the inflammation response.

18.
Article in Chinese | WPRIM | ID: wpr-905901

ABSTRACT

Objective:To explore the effect and underlying mechanism of koumine (Kou) at different concentrations (0, 100, 200, 400 μmol·L<sup>-1</sup>) on the proliferation and apoptosis of colorectal cancer HCT-116 cells. Method:After 24 hours of<italic> in vitro</italic> intervention with HCT-116 cells by Kou, cell counting kit-8 (CCK-8) assay was used to detect its effect on cell proliferation. Flow cytometry was used to detect cell cycle, apoptosis, and reactive oxygen species (ROS) expression. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of forkhead box O3a (FoxO3a). Cells were transfected with small interfering ribonucleic acid (siRNA). Western blot was employed to detect the protein expression of the FoxO3a target gene. Result:Compared with the conditions in the blank group, Kou treatment reduced the proliferation rate of HCT-116 cells (<italic>P</italic><0.05, <italic>P</italic><0.01) in a dose-dependent manner, caused cell cycle arrest in the G<sub>0</sub>/G<sub>1</sub> phase, and induced the apoptosis of HCT-116 cells (<italic>P</italic><0.05, <italic>P</italic><0.01), which was positively correlated with the concentration of Kou. FoxO3a siRNA interference reduced the expression of FoxO3a and its downstream target genes cyclin-dependent kinase inhibitor 1A (p21), cyclin-dependent kinase inhibitor 1B (p27), and Bcl-2 interacting mediator of cell death (Bim) (<italic>P</italic><0.01). Kou treatment induced the activation of c-Jun <italic>N</italic>-terminal kinase (JNK) in HCT116 cells. SP600125 (JNK specific inhibitor) treatment inhibited the Kou-induced FoxO3a activation and the expression of its downstream target genes. <italic>N</italic>-acetyl cysteine (NAC) treatment reduced Kou-induced ROS levels (<italic>P</italic><0.01) and JNK signal activation. The above results were significantly different from those in the blank group (<italic>P</italic><0.01). Conclusion:Kou can effectively inhibit the proliferation of HCT-116 cells and promote apoptosis, and the mechanism may be related to the regulation of the ROS/JNK/FoxO3a pathway.

19.
Acupuncture Research ; (6): 21-26, 2020.
Article in Chinese | WPRIM | ID: wpr-844211

ABSTRACT

OBJECTIVE: To observe the effect of electroacupuncture (EA) stimulation on the expression of c-Jun terminal kinase(JNK)signaling pathway-related proteins in the hippocampus of vascular dementia (VD) rats, so as to explore its mechanisms underlying improvement of VD. METHODS: Male Sprague-Dawley rats were randomly divided into sham operation, model and EA groups (n=10 rats per group). The VD model was prepared by repeated occlusion of the bilateral common carotid arteries for 10 min and reperfusion for 10 min (3 times in total). The rats in the EA group received EA (2 Hz, 2 mA) at "Dazhui"(GV14),"Baihui"(GV20), and bilateral "Housanli"(ST36) ,"Geshu"(BL17) for 10 min, once daily for 14 days. The learning-memory abi-lity was detected by Morris water maze tests, the distribution of hippocampal neurons detected by Nissl staining, and the apoptosis of hippocampal neurons detected by using TdT-mediated dUTP nick-end labeling (TUNEL) method. The expressions of JNK, phosphorylated JNK (p-JNK), cysteine-containing aspartate-specific proteases-8 (Caspase-8) and Caspase-3 proteins were detected by Western blot. RESULTS: After modeling and compared with the sham operation group, the escape latency was significantly prolonged (P<0.01) and the number of safe-platform quadrant crossing obviously decreased (P<0.01), suggesting a reduction of learning-memory ability. The number of hippocampal neurons was considerably reduced (P<0.01), and that of hippocampal apoptotic neurons remarkably increased in the model group (P<0.01). Whereas, the expression levels of hippocampal apoptosis-related proteins as JNK, p-JNK, Caspase-8 and Caspase-3, as well as the apoptotic index were significantly up-regulated (P<0.01). Following EA intervention, the learning-memory ability was apparently improved (P<0.01), and the number of hippocampal neurons was considerably increased (P<0.01), the hippocampal apoptotic cell number, apoptosis index and the expression levels of JNK, p-JNK, Caspase-8 and Caspase-3 were significantly down-regulated (P<0.01). CONCLUSION: EA intervention can improve the learning-memory ability of VD rats, which may be associated with its effects in reducing hippocampal apoptosis by suppressing JNK signaling pathway.

20.
Article in Chinese | WPRIM | ID: wpr-873016

ABSTRACT

Objective:To investigate the effect of ginkgolide B (GB) on the activation of c-Jun aminoterminal kinase(JNK) signaling pathway and apoptosis in amyotrophic lateral sclerosis cell model. Method:NSC34 cells were infected by slow virus containing expression superoxide dismutase1(SOD1)WT and hSOD1G93A and empty plasmid, and screened with a certain concentration of puromycin, so as to observe the transfection efficiency of slow virus and cell morphology under inverted fluorescence microscope. Western blot method was used to verify whether infected cells were over-expressing SOD1 target proteins. The hSOD1G93A-NSC34 cell lines were established and given GB. Cell cultures were divided into normal group, model group and different concentrations of ginkgolide B groups (25, 50, 75, 100 mg∙L-1). After 48 h, methyl thiazolyl tetrazolium (MTT) was used to detect cell survival rates, and select the best drug concentration. Subsequent experimental groups were divided into normal group, model group, 75 mg∙L-1 GB group, SP600125 group, and 75 mg∙L-1 GB + SP600125 group. Flow cytometry was used to detect the apoptosis of each group of cells. Western blot was used to detect the expressions of phosphorylation(p)-JNK, c-Jun, p-c-Jun, and cysteine aspartic acid protease -3(Caspase-3) proteins. Result:Compared with normal NSC34 cells, hSOD1G93A-NSC34 cell body became round, the synapses decreased and shortened, but the cell morphology of hSODWT-NSC34 cell and empty plasmid group did not change significantly. Western blot showed that hSOD1G93A-NSC34, hSOD1WT-NSC3 intracellular SOD1 protein levels increased significantly (P<0.01), and the amyotrophic lateral sclerosis cell model was established. Compared with the normal group, the cell activity in the model group was significantly reduced (P<0.01). Compared with the model group, the cell activity increased at different concentrations of GB, especially when the drug concentration was 75 mg∙L-1 (P<0.01). In subsequent experiments, compared with the normal group, the apoptosis, and expressions of p-JNK, p-c-Jun, and cleaved Caspase-3 proteins in the model group increased significantly (P<0.01). Compared with the model group, the apoptosis and p-JNK, p-c-Jun, released Caspase-3 protein expressions of 75 mg∙L-1 GB group, SP600125 group, 75 mg∙L-1 GB + SP600125 group decreased significantly (P<0.05, P<0.01). Conclusion:GB has a protective effect on the cell model of atrophy lateral sclerosis, which may be realized by JNK signal pathway.

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