Angelica polymorpha Maxim Induces Apoptosis of Human SH-SY5Y Neuroblastoma Cells by Regulating an Intrinsic Caspase Pathway

Rahman, Md. Ataur,Bishayee, Kausik,Huh, Sung-Oh Korean Society for Molecular and Cellular Biology 2016 Molecules and cells Vol.39 No.2

Angelica polymorpha Maxim root extract (APRE) is a popular herbal medicine used for treating stomachache, abdominal pain, stomach ulcers, and rheumatism; however the effect of APRE on cancer cells has not yet been explored. Here, we examined APRE cytotoxicity seen on target neuroblastoma cells (NB) using cell viability assays, DAPI visualization of fragmented DNA, and Western blotting analysis of candidate signaling pathways involved in proliferation and apoptosis. We demonstrated that APRE reduced cell viability in NB to a greater extent than in fibroblast cells. In addition, we found that APRE could inhibit the three classes of MAPK proteins and could also down-regulate the PI3K/AKT/GSK-$3{\beta}$ activity all being relevant for proliferation and survival. APRE could also up-regulate Bax expression and down-regulate Bcl-2 and Mcl-1. With APRE treatment, depolarization of mitochondria membrane potential and activation of caspase-3 was demonstrated in the SH-SY5Y cells. We could not found increased activity of death receptor and caspase-8 as markers of the extrinsic apoptosis pathway for the APRE treated cells. In presence of a caspase-3 siRNA and a pan-caspase inhibitor, APRE could not reduce the viability of NB cells to a significant degree. So we predicted that with APRE, the intrinsic pathway was solely responsible for inducing apoptosis as we also showed that the non-caspase autophagy pathway or ER stress-ROS mediated pathways were not involved. These findings demonstrate that an intrinsic mitochondria-mediated apoptosis pathway mediates the apoptotic effects of APRE on SH-SY5Y cells, and that APRE shows promise as a novel agent for neuroblastoma therapy.

Angelicin induces apoptosis through intrinsic caspase-dependent pathway in human SH-SY5Y neuroblastoma cells

Ataur Rahman, Md.,Kim, Nam-Ho,Yang, Haijie,Huh, Sung-Oh Springer-Verlag 2012 MOLECULAR AND CELLULAR BIOCHEMISTRY - Vol.369 No.1

<P>Angelicin is structurally related to psoralens, a well-known chemical class of photosensitizers used for its antiproliferative activity in treatment of different skin diseases. To verify the activity of angelicin, we employed human SH-SY5Y neuroblastoma cells to investigate its cytotoxicity, although its mechanism of action has not yet been fully elucidated. Here, we examined the cellular cytotoxicity of angelicin by cell viability assay, DNA fragmentation by DNA ladder assay, and activation of caspases and Bcl-2 family proteins by western blot analyses. The results of our investigation suggest that angelicin increased cellular cytotoxicity in a dose- and time-dependent manner with IC50 of 49.56 mu M at 48 h of incubation. In addition, angelicin dose-dependently downregulated the expression of anti-apoptotic proteins including Bcl-2, Bcl-xL, and Mcl-1 suggesting the involvement of the intrinsic mitochondria-mediated apoptotic pathway which did not participate in Fas/FasL-induced caspase-8-mediated extrinsic, MAP kinases, and PI3K/AKT/GSK-3 beta pathway. Furthermore, we clarified the dose-dependent upregulation of caspase-9 and caspase-3 which indicated that angelicin-induced apoptosis is mediated primarily through the intrinsic caspase-mediated pathway. In particular, the caspase-3 inhibitor, DEVD-fmk, induced a reduction in angelicin-induced cytotoxicity which confirmed that the intrinsic caspase-dependent pathway during this apoptosis which did not prevent cytotoxicity using MAP kinases and GSK-3 inhibitor. Taken together, our data shows that angelicin is an effective apoptosis-inducing natural compound of human SH-SY5Y neuroblastoma cells which suggests that this compound may have a role in future therapies for human neuroblastoma cancer.</P>

Cytotoxic effect of gambogic acid on SH-SY5Y neuroblastoma cells is mediated by intrinsic caspase-dependent signaling pathway.

Rahman, Md Ataur,Kim, Nam-Ho,Huh, Sung-Oh Dr. W. Junk B. V. Publishers ; Kluwer Academic Pub 2013 MOLECULAR AND CELLULAR BIOCHEMISTRY - Vol.377 No.1

<P>Gambogic acid (GA) is the dry resin of Garcinia hanburyi (Guttiferae) with potent anti-tumor activity, various bioactivities, including detoxification, homeostasis, anti-inflammatory, and parasiticide, whereas the effect of this natural compound on cancer cells has not been clearly clarified. Here, we examined cellular cytotoxicity by cell viability assay and DNA fragmentation by DNA-ladder assay. Activation of different protein expressions were detected by western blot analyses. We first demonstrated that GA reduces the human SH-SY5Y neuroblastoma cell viability with IC50 of 1.28 μM at 6 h which has less toxicity in fibroblast cells. However, lower concentration GA significantly downregulated the expression of anti-apoptotic protein including Bcl-2, Bcl-xL, and Mcl-1, which also dramatically activated cleaved caspase-9 and -3 in a dose- and time-dependent manner. Consequently, GA-induced cytotoxicity was not mediated by the Fas/FasL and PI3 K/AKT/GSK-3β signaling pathway. In addition, GA-induced cells showed damage morphology which had become cell rounding, neurite retraction, membrane blebbing and shrunken in a dose- and time-dependent manner that clearly indicates this morphological change might be due to the process of apoptosis which shows fragmented DNA. Therefore, the findings presented in this study demonstrate that apoptotic effects of GA on SH-SY5Y cells are mediated by intrinsic mitochondrion-dependent caspase pathway which suggests this natural compound might be effective as an anti-cancer agent for neuroblastoma malignancies.</P>

Energy efficient electromagnetic actuated CVT system

Ataur Rahman,Sazzad Bin Sharif,A.K.M. Mohiuddin,Mahbubur Rashid,Altab Hossain 대한기계학회 2014 JOURNAL OF MECHANICAL SCIENCE AND TECHNOLOGY Vol.28 No.4

A continuously variable transmission (CVT) system transmits the engine/battery power to the car driving wheel smoothly and efficiently. Several types of CVT already been developed to improve the transmission losses while maintaining acceleration time. However,most of the CVT has some constraints in the actuation mechanism which led us to develop an innovative electromagnetic actuator forCVT. Simplified mathematical equations have been developed for the kinematics analysis of clamping forces of the CVT and electromagneticforces of EMA. The EMA has been developed for ¼ scale car with two sets of solenoid. Each of the two sets has been equippedwith primary and secondary pulleys for pushing and pulling the movable sheave. The solenoid is operated by controlling the supply currentwith a fuzzy logic controller. A simulation based fuzzy logic controller has been introduced here for identifying the desired current ofthe EMA actuation. The experimental results show that the EMA develops electromagnetic forces 301 N for the supply current of 3.37amp, which makes the acceleration time of the car in the range of 2.5~3.5 sec and electromagnetic actuated CVT system highly energyefficient.

Antiproliferative and Cytotoxic Effects of Resveratrol in Mitochondria-Mediated Apoptosis in Rat B103 Neuroblastoma Cells

Rahman, Md. Ataur,Kim, Nam-Ho,Kim, Seung-Hyuk,Oh, Sung-Min,Huh, Sung-Oh The Korean Society of Pharmacology 2012 The Korean Journal of Physiology & Pharmacology Vol.16 No.5

Resveratrol, a natural compound, has been shown to possess anti-cancer, anti-aging, anti-inflammatory, anti-microbial, and neuroprotective activities. In this study, we examined the antiproliferative and cytotoxicity properties of resveratrol in Rat B103 neuroblastoma cells; although it's molecular mechanisms for the biological effects are not fully defined. Here, we examined the cellular cytotoxicity of resveratrol by cell viability assay, antiproliferation by BrdU assay, DNA fragmentation by DNA ladder assay, activation of caspases and Bcl-2 family proteins were detected by western blot analyses. The results of our investigation suggest that resveratrol increased cellular cytotoxicity of Rat B103 neuroblastoma cells in a dose-and time-dependent manner with $IC_{50}$ of 17.86 ${\mu}M$ at 48 h. On the other hand, incubation of neuroblastoma cells with resveratrol resulted in S-phase cell cycle arrests which dose-dependently and significantly reduced BrdU positive cells through the downregulation of cyclin D1 protein. In addition, resveratrol dose-dependently and significantly downregulated the expression of anti-apoptotic protein includes Bcl-2, Bcl-xL and Mcl-1 and also activates cleavage caspase-9 and-3 via the downregulation of procaspase-9 and -3 in a dose-dependent manner which indicates that involvement of intrinsic mitochondria-mediated apoptotic pathway. In conclusion, resveratrol increases cellular cytotoxicity and inhibits the proliferation of B103 neuroblastoma cells by inducing mitochondria-mediated intrinsic caspase dependent pathway which suggests this natural compound could be used as therapeutic purposes for neuroblastoma malignancies.

TWO-PHASE EVAPORATIVE BATTERY THERMAL MANAGEMENT TECHNOLOGY FOR EVs/HEVs

Rahman Ataur,Mohammed Nurul Amin Hawlader,Helmi Khalid 한국자동차공학회 2017 International journal of automotive technology Vol.18 No.5

Electric vehicle’s motor draws power from battery to meet its power demand in different road profiles. Battery high discharged currents are causes of warming battery’s cells. The temperature of 40 ºC and above reduces battery life span. The rationale of fuzzy controlled evaporative battery thermal management system (EC-BThMS) development from this study is to control the battery temperature in the range of 20 ~ 40 ºC both in charging/discharging modes. The proposed system has been developed with estimating the total cooling loads and thermal behavior of the battery cells. A fuzzy controlling system has been introduced with the EC-BThMS to control the electro-compressor and the expansion valve based on the response of battery temperature sensors.A battery pack of 8.6 kWh equipped EV has been operated with 60 km/h on 0 % gradient and 40 km/h on 5 % gradient in IIUM campus while 130 km/h on 0 % gradient and 50 km/h on 3.67 % gradient in Malaysia International Formula circuit to study the battery temperature profile and percentage of battery power saving. Comparison has been made on the performance of EC-BThMS with air cooling battery thermal management system (AC-BThMS) by using same vehicle. Result shows that EC-BThMS can save energy 17.69 % more than AC-BThM 1 and 23 % more than AC-BThM 2.

Gintonin stimulates autophagic flux in primary cortical astrocytes

Rahman, Md. Ataur,Hwang, Hongik,Nah, Seung-Yeol,Rhim, Hyewhon The Korean Society of Ginseng 2020 Journal of Ginseng Research Vol.44 No.1

Background: Gintonin (GT), a novel ginseng-derived exogenous ligand of lysophosphatidic acid (LPA) receptors, has been shown to induce cell proliferation and migration in the hippocampus, regulate calcium-dependent ion channels in the astrocytes, and reduce β-amyloid plaque in the brain. However, whether GT influences autophagy in cortical astrocytes is not yet investigated. Methods: We examined the effect of GT on autophagy in primary cortical astrocytes using immunoblot and immunocytochemistry assays. Suppression of specific proteins was performed via siRNA. LC3 puncta was determined using confocal microscopy. Results: GT strongly upregulated autophagy marker LC3 by a concentration- as well as time-dependent manner via G protein-coupled LPA receptors. GT-induced autophagy was further confirmed by the formation of LC3 puncta. Interestingly, on pretreatment with an mammalian target of rapamycin (mTOR) inhibitor, rapamycin, GT further enhanced LC3-II and LC3 puncta expression. However, GT-induced autophagy was significantly attenuated by inhibition of autophagy by 3-methyladenine and knockdown Beclin-1, Atg5, and Atg7 gene expression. Importantly, when pretreated with a lysosomotropic agent, E-64d/peps A or bafilomycin A1, GT significantly increased the levels of LC3-II along with the formation of LC3 puncta. In addition, GT treatment enhanced autophagic flux, which led to an increase in lysosome-associated membrane protein 1 and degradation of ubiquitinated p62/SQSTM1. Conclusion: GT induces autophagy via mTOR-mediated pathway and elevates autophagic flux. This study demonstrates that GT can be used as an autophagy-inducing agent in cortical astrocytes.

5-Hydroxytryptamine 6 Receptor (5-HT₆R)-Mediated Morphological Changes via RhoA-Dependent Pathways

Rahman, Md. Ataur,Kim, Hanna,Lee, Kang Ho,Yun, Hyung-Mun,Hong, Jung-Hwa,Kim, Youngjae,Choo, Hyunah,Park, Mikyoung,Rhim, Hyewhon Korean Society for Molecular and Cellular Biology 2017 Molecules and cells Vol.40 No.7

The $5-HT_6R$ has been considered as an attractive therapeutic target in the brain due to its exclusive expression in the brain. However, the mechanistic linkage between $5-HT_6Rs$ and brain functions remains poorly understood. Here, we examined the effects of $5-HT_6R$-mediated cell morphological changes using immunocytochemistry, Western blot, and live-cell imaging assays. Our results showed that the activation of $5-HT_6Rs$ caused morphological changes and increased cell surface area in HEK293 cells expressing $5-HT_6Rs$. Treatment with 5-HT specifically increased RhoA-GTP activity without affecting other Rho family proteins, such as Rac1 and Cdc42. Furthermore, live-cell imaging in hippocampal neurons revealed that activation of $5-HT_6Rs$ using a selective agonist, ST1936, increased the density and size of dendritic protrusions along with the activation of RhoA-GTP activity and that both effects were blocked by pretreatment with a selective $5-HT_6R$ antagonist, SB258585. Taken together, our results show that $5-HT_6R$ plays an important role in the regulation of cell morphology via a RhoA-dependent pathway in mammalian cell lines and primary neurons.

Kinematics and nonlinear control of an electromagnetic actuated CVT system for passenger vehicle

Ataur Rahman,Sazzad Bin Sharif,Altab Hossain,A. K. M. Mohiuddin,A. H. M. Zahirul Alam 대한기계학회 2012 JOURNAL OF MECHANICAL SCIENCE AND TECHNOLOGY Vol.26 No.7

An electromagnetic actuated continuously variable transmission (EMA-CVT) system is developed by two sets of electromagnetic actuators (solenoid) located on primary and secondary pulley. A set of solenoids are attached to the primary and secondary pulley to develop the attraction and repulsive forces. The relationships between the speed ratio and electromagnetic actuation and clamping force and output torque of the CVT are established based on the kinematics of the EMA-CVT system. A fuzzy logic controller (FLC) is developed to control the EMA precisely based on the feedback of the RPM sensor and slope sensor. The EMA-CVT performance with controller has found 28% more than the performance of the EMA-CVT without controller. The solenoids of the EMA were activated by varying the current supply with the Fuzzy-Proportional-Derivative-Integrator (FPID) to maintain the non-linearity of the CVT in response of the vehicle traction torque demand. Result shows that the solenoid is able to pull the plunger in the desired distance with supply current of 12.5 amp while push the plunger to the desired distance with 14.00 amp current supply to the windings when the vehicle is considered in 10% grade. The acceleration time of the ¼ scale car has been recorded as 5.5 s with the response of drive wheels torque.

Induction of apoptosis by Dioscorea nipponica Makino extracts in human SH-SY5Y neuroblastoma cells via mitochondria-mediated pathway

Rahman, Md. Ataur,Yang, Haijie,Kim, Nam-Ho,Huh, Sung-Oh 한국통합생물학회 2014 Animal cells and systems Vol.18 No.1

Dioscorea nipponica, a perennial herb growing in mountainous areas, has been used as a folk medicine for asthma, rheumatoid arthritis, bronchitis, and other disease in Korea, whereas the effect of this plant on cancer cells has not been clearly clarified. Cellular cytotoxicity was examined by cell viability assay and DNA fragmentation by DNA ladder assay. Activation of different protein expression was detected by western blot analyses. We first demonstrated that D. nipponica extract (DNE) reducing the SH-SY5Y cell viability with $IC_{50}$ of $27.57{\mu}g/mL$ at 24 h. However, DNE downregulated the expression of antiapoptotic protein includes B-cell lymphoma-2 (Bcl-2), B-cell lymphoma-extra large (Bcl-xL), and myeloid cell leukemia 1 (Mcl-1) which also activated cleaved caspase-9 and caspase-3 in a dose- and time-dependent manner. Consequently, DNE-induced cytotoxicity was not mediated by the Fas/FasL, phosphatidylinositide-3 kinase/AKT/glycogen synthase kinase-$3{\beta}$ (PI3K/AKT/GSK-$3{\beta}$), and mitogen activated protein (MAP) kinases signaling pathway. In addition, DNE-induced cells shown damage morphology which had become cell rounding, neurite retraction, membrane blebbing, is right spell blebbing, and shrunken in a dose- and time-dependent manner that clearly indicate this morphological change might be due to the process of apoptosis which shown fragmented DNA. These results indicate that apoptotic effects of DNE on SH-SY5Y cells are mediated by intrinsic mitochondrial caspases signaling pathway, suggesting that DNE might be effective as an anticancer agent for neuroblastoma malignancies.