Contractile and Electrical Responses of Guinea-pig Gastric Smooth Muscle to Bradykinin

Kim. Chul-Soo,Jun. Jae-Yeoul,Kim. Sung-Joon,So. In-Suk,Kim. Ki-Whan 대한생리학회 1995 대한생리학회지 Vol.29 No.2

The nonapeptide bradykinin has been shown to exhibit an array of biological activities including relaxation/contraction of various smooth muscles. In order to investigate the effects of bradykinin on the contractility and the electrical activity of antral circular muscle of guinea-pig stomach, the isometric contraction and membrane potential were recorded. Also, using standard patch clamp technique, the Ca²⁺-activated K currents were recorded to observe the change in cytosolic Ca²⁺ concentration. 0.4 μM bradykinin induced a triphasic contractile response (transient contraction-transient relaxation-sustained contraction) and this response was unaffected by pretreatment with neural blockers (tetrodotoxin, atropine and guanethidine) or with apamin. Bradykinin induced hyperpolarization of resting membrane potential and enhanced the amplitude of slow waves and spike potentials. The enhancement of spike potentials was blocked by neural blockers. Both the bradykinin-induced contractions and changes in membrane potential were reversed by the selective B₂-receptor antagonist (N α-adamantaneacetyl-_D-Arg-[Hyp, Thy,_D-Phe]-bradykinin). In whole-cell patch clamp experiment, we held the membrane potential at -20 mV and spontaneous and transient changes of Ca-activated K currents were recorded. Bradykinin induced a large transient outward current, consistent with a calcium-releasing action of bradykinin front the intracellular calcium pool, because such change was blocked by pretreatment with caffeine. Bradykinin-induced contraction was also blocked by pretreatment with caffeine. From these results, it is suggested that bradykinin induces a calciumrelease and contraction through the B₂ receptor of guinea-pig gastric smooth muscle. Enhancement of slow wave activity is an indirect action of bradykinin through enteric nerve cells embedded in muscle strip.

흰쥐에서 측뇌실내 Bradykinin의 승압효과

김장만,염철호 朝鮮大學校 附設 醫學硏究所 1992 The Medical Journal of Chosun University Vol.17 No.2

Central hemodynamic actions of bradykinin were studied in anesthetized rats in order to investigate its mode of action in the central nervous system. Bradykinin(0.5~10 nmoℓ) was administered into the right lateral ventricle, while blood pressure and heart rate were monitored continuously. Effects of various drugs including phentolamine(2 ㎎/㎏), hexamethonium(2.5 ㎎/㎏/min), reserpine (2 ㎎/㎏/day), indomethacin(10 ㎎/㎏) and sara1asin(20 ㎍/㎏/min) on the hemodynamic effects of bradykinin were also examined. The results obtained were as follows: 1. Administration of bradykinin into the lateral ventricle, elicited an increase in mean arterial pressure without significant changes in heart rate. 2. Bradykinin, doses ranging from 0.5 to 10 nmoℓ, produced a dose-dependent increase in mean arterial pressure. The maximum increase in blood pressure was noted at 5 nmoℓ, 3. Central pressor responses to bradykinin were attenuated by pretreatment with phentolamine or hexamethonium. Reserpine, indomethacin and saralasin did not affect the pressor response to bradykinin. 4. These results suggest that the central pressor effect of bradykinin is, at least in part, due to excitation of the autonomic nervous activity. Mechanisms other than the enhanced sympathetic nervous activity cannot be ruled out completely in the central action of bradykinin.

Anti-nociceptive effect of Bee Venom on Capsaicin or Bradykinin-induced Pain

Yang Chang-Yeol,Song Ho-sueb 대한침구의학회 2007 대한침구의학회지 Vol.24 No.2

연구목적 : 봉독으로 유발된 통각수용의 강도와 봉독으로 나타나는 항통각수용(통각억제성)의 강도를 쥐의 포르말린 테스트를 통해 상호관련됨을 확인하고 capsaicin과 bradykinin으로 통증 유발된 쥐의 자발적인 통증행동(핥기횟수측정; LN), 꼬리경타시험(TFL)과 열판시험(HPL)을 통해 봉독의 항통각수용(통각억제)작용을 재확인 하고자 하였다. 실험방법 : 쥐의 뒷다리에 통증유도 물질인 Capsaicin 또는 Bradykinin을 20㎕를 주사하여 동통을 유발하고 자발적 통증행동인 핥기횟수측정(LN), 꼬리경타기간(TFL)과 열판위에서의 온도자극에 쥐가 반응하는 시간을 측정(HPL)하는 실험을 봉독주입하거나 몰핀을 주입하거나 아무것도 주입하지 않고 통증유발만 시킨 이후에 각각 시행하였다. 결 과: 1. Capsaicin 또는 Bradykinin으로 동통유발 후 LN은 두드러진 증가를 보임, HPL은 감소를 TFL은 두드러진 감소를 나타내었다. 2. 봉독이나 몰핀 주입 30분후에 Capsaicin으로 동통유발이후 LN은 봉독과 몰핀에서 모두 현격한 감소를 HPL은 봉침은 현격한 증가를 몰핀에서는 감소를 TFL은 봉침과 몰핀에서 모두 현격한 증가를 나타내었다. 3. 봉독과 몰핀 주입 30분후에 Bradykinin으로 동통유발이후 LN은 봉독은 증가 몰핀은 현격한 감소를 HPL은 봉침은 증가 몰핀에서는 현격한 증가를 TFL은 봉침과 몰핀에서 모두 증가를 나타내었다. 결 론 : 봉독은 Capsaicin 또는 Bradykinin으로 동통유발된 통각수용 행동을 감소시키는 결과를 나타내었는데 이것은 기존의 연구결과들에서의 봉독의 항통각수용(통각억제성)의 효과를 입증하였고 봉약침은 염증의 개선이나 암과 관련된 동통에 유효한 방법임을 시사하는 것이다. Objectives : To confirm whether the intensity of the BV-induced nociception is correlated with the intensity of the BV-induced antinociception in the mouse formalin test(as measured by spontaneous pain behaviour).To reconfirm the anti-nociceptive effect of BV, we evaluated anti-nociceptive effect of BV on capsaicin or bradykinin-induced pain behaviors of rats by spontaneous pain behaviour(lick), tail flick latency(TFL) and hot plate test.Material and Methods : 20 ㎕ and 0.5 mg/Kg of BV or 30 ㎕ and 10 mg/Kg of morphine was injected into a hind paw of mice intraplantarly 30 minutes before pain was induced by 10 nmol of capsaicin or 20 nmol of bradykinin. and we evaluated anti-nociceptive effect of BV on capsaicin or bradykinin-induced pain behaviors of rats by spontaneous pain behaviour(licking number:LN), tail flick latency(TFL) and hot plate test(HPL).

Central Pressor Mechanisms of Bradykinin in 2-Kidney, 1 Clip Goldblatt Hypertensive Rats

Yeum, Cheol-Ho,Jun, Jae-Yeoul,Yoon, Pyung-Jin The Korean Physiological Society 1992 대한생리학회지 Vol.26 No.1

Central cardiovascular effects of bradykinin were examined in anesthetized normotensive (NTR) and 2-kidney, 1 clip Goldblatt hypertensive rats (GHR). Bradykinin ($0.5{\sim}10nmol$) was administered into the right lateral cerebral ventricle, while blood pressure and heart rate (HR) were continuously monitored. In both NTR and GHR, intracerebroventricular bradykinin produced a dose dependent increase in mean arterial pressure (MAP) without significant changes in HR. GHR were more sensitive in the pressor response than NTR. The pressor response to bradykinin was attenuated by treatment with hexamethonium (2.5mg/kg/min, IV) or phentolamine (2mg/kg, IV) in both NTR and GHR. Reserpine treatment (2mg/kg/day, intramuscularly,2 days) did not affect the central pressor effect of bradykinin in NTR but it attenuated the pressor effect in GHR. Pretreatment with indomethacin (10mg/kg, intraperitoneally) or saralasin ($20{\mu}g$/kg/min, IV) was without effects on the pressor response to bradykinin. These results indicate that the central pressor effect of bradykinin is, at least in part, due to excitation of the autonomic nervous activity. Mechanisms other than the enhanced sympathetic nervous activity ran. not be ruled out, However. It is also suggested that the sensitivity to bradykinin is increased in the GHR.

Adenoviral-Mediated Ref-1 Overexpression Potentiates NO Production in Bradykinin-Stimulated Endothelial Cells

송주동,김강미,이상권,김종민,박영철,Song, Ju-Dong,Kim, Kang-Mi,Lee, Sang-Kwon,Kim, Jong-Min,Park, Young-Chul Korean Society of Life Science 2007 생명과학회지 Vol.17 No.7

Redox Factor-1 (Ref-1)은 손상된 DNA의 복구 및 많은 세포내 산화환원에 민감한 transcription factors의 활성화에 기여하는 양면의 역할을 수행하는 단백질이다. 본 연구에서는 혈관내피세포에서의 nitric oxide (NO) 생성과정에서 Ref-1의 역할을 살펴보았다. Ref-1의 세포내 과발현을 위하여 adenoviral vector를 사용하였고 bradykinin으로 자극한 혈관내피세포에서 생성되는 NO 측정을 위하여 fluorophore DAF-2를 사용하였다. Ref-1 과 발현은 bradykinin으로 자극한 혈관내피세포의 NO 생성을 증가시켰다. 또한 자극되지 않은 Ref-1 과발현 세포는 viral vector로 감염되지 않은 그리고 control로 사용한 AdD1312로 감염된 세포보다 높은 fluorescence intensity를 나타내었다. 이와 비슷하게, Ref-1 과발현은 bradykinin으로 자극한 세포뿐만 아니라 자극하지 않은 세포에서도 감염되지 않은 그리고 AdD1312로 감염된 세포와 비교할 때 endothelial NO synthase (eNOS)의 활성을 크게 증가시켰다. 이는Ref-1 자신이 eNOS의 효소활성을 직접 조절할 수 있다는 것을 의미한다. 결론적으로 Ref-1이 혈관계에서 NO생성에 의해 기인되는 endothelium-dependent vasorelaxation에서 중요한 역할을 한다는 것을 시사한다. The dual-function protein redox factor-1 (Ref-1) is essential for base excision repair of oxidatively damaged DNA and also governs the activation of many redox-sensitive transcription factors. We examined the role of Ref-1 in regulation of nitric oxide (NO) synthesis employing adenoviral-mediatedoverexpression of Ref-1 in bradykinin-stimulated endothelial cells. Intracellular NO was detected with the NO-sensitive fluorophore DAF-2. Overexpression of Ref-1 potentiates bradykinin-stimulated NO production in endothelial cells. And, cells ifected with AdRef-1 showed higher fluorescence intensity compared with uninfected or AdD1312-infected cells. In parallel with this, over expression of Ref-1 also stimulated endothelial NO synthase (eNOS) enzyme activity, compared with unifected or AdD1312-infected cells, in bradykinin-stimulated cells as well as in unstimulated cells. These results suggest that Ref-1 implicates in endothelium-dependent vasorelaxation resulting from NO production in vascular system.

Bradykinin-induced Ca²^(+) signaling in human oral squamous cell carcinoma HSC-3 cells

Sohn, Byung Jin,Kang, Ji-Ah,Jo, Su-Hyun,Choi, Se-Young KOREAN ACADAMY OF ORAL BIOLOGY 2009 International Journal of Oral Biology Vol.34 No.2

Cytosolic Ca²^(+) is an important regulator of tumor cell proliferation and metastasis. Recently, the strategy of blocking receptors and channels specific to certain cancer cell types has emerged as a potentially viable future treatment. Oral squamous cell carcinoma is an aggressive form of cancer with a high metastasis rate but the receptor-mechanisms involved in Ca²^(+) signaling in these tumors have not yet been elucidated. In our present study, we report that bradykinin induces Ca²^(+) signaling and its modulation in the human oral squamous carcinoma cell line, HSC-3. Bradykinin was found to increase the cytosolic Ca²^(+) levels in a concentration-dependent manner. This increase was inhibited by pretreatment with the phospholipase C-β inhibitor, U73122, and also by 2-aminoethoxydiphenyl borate, an inhibitor of the inositol 1,4,5-trisphosphate receptor. Pretreatment with extracellular ATP also inhibited the peak bradyknin-induced Ca²^(+) rise. In contrast, the ATP-induced rise in cytosolic Ca²^(+) was not affected by pretreatment with bradykinin. Pretreatment of the cells with either forskolin or phorbol 12-myristate 13-acetate(activators of adenylyl cyclase and protein kinase C, respectively) prior to bradykinin application accelerated the recovery of cytosolic Ca²^(+) to baseline levels. These data suggest that bradykinin receptors are functional in Ca²^(+) signaling in HSC-3 cells and may therefore represent a future target in treatment strategies for human oral squamous cell carcinoma.

Depolarizing Effectors of Bradykinin Signaling in Nociceptor Excitation in Pain Perception

Choi, Seung-In,Hwang, Sun Wook The Korean Society of Applied Pharmacology 2018 Biomolecules & Therapeutics(구 응용약물학회지) Vol.26 No.3

Inflammation is one of the main causes of pathologic pain. Knowledge of the molecular links between inflammatory signals and pain-mediating neuronal signals is essential for understanding the mechanisms behind pain exacerbation. Some inflammatory mediators directly modulate the excitability of pain-mediating neurons by contacting the receptor molecules expressed in those neurons. For decades, many discoveries have accumulated regarding intraneuronal signals from receptor activation through electrical depolarization for bradykinin, a major inflammatory mediator that is able to both excite and sensitize pain-mediating nociceptor neurons. Here, we focus on the final effectors of depolarization, the neuronal ion channels, whose functionalities are specifically affected by bradykinin stimulation. Particular G-protein coupled signaling cascades specialized for each specific depolarizer ion channels are summarized. Some of these ion channels not only serve as downstream effectors but also play critical roles in relaying specific pain modalities such as thermal or mechanical pain. Accordingly, specific pain phenotypes altered by bradykinin stimulation are also discussed. Some members of the effector ion channels are both activated and sensitized by bradykinin-induced neuronal signaling, while others only sensitized or inhibited, which are also introduced. The present overview of the effect of bradykinin on nociceptor neuronal excitability at the molecular level may contribute to better understanding of an important aspect of inflammatory pain and help future design of further research on the components involved and pain modulating strategies.

Effects of Intracerebroventricular Captopril on the Central Pressor Response to Bradykinin in Normotensive and Hypertensive Rats

Yeum, Cheol-Ho,Yoon, Pyung-Jin The Korean Physiological Society 1994 대한생리학회지 Vol.28 No.2

Captopril, an inhibitor of angiotensin converting enzyme, is also known to inhibit the degradation of bradykinin. We examined the effects of intracerebroventricular (ICV) captopril on the central pressor response to bradykinin in normotensive, 2-kidney, 1 clip Goldblatt (GHR) and deoxycorticosterone acetate (DOCA)-salt hypertensive rats. Captopril (1 mg) and bradykinin (5 nmol) were administered into the right lateral cerebral ventricle, and blood pressure and heart rate were continuously monitored throughout the experiment. ICV captopril alone did not affect the blood pressure within 10 minutes but it significantly augmented the central pressor response to bradykinin in GHR. On the contrary, captopril was without effect on the pressor response to bradykinin in normotensive and DOCA-salt rats. These findings indicate that endogenous kinins are not critical in regulating arterial pressure in normotensive and DOCA hypertensive rats. However, in GHR, an enhanced activity of the brain kallikrein-kinin system in maintaining the high blood pressure is suggested.

Comparison of Bradykinin- and Platelet-Derived Growth Factor-Induced Phosphoinositide Turnover in NIH 3T3 Cells

Lee, Kee-Ho,Ryu, Yong-Wun,Yoo, Young-Do,Bai, Dong-Hoon,Yu, Ju-Hyun,Kim, Chang-Min Korean Society for Biochemistry and Molecular Biol 1996 Journal of biochemistry and molecular biology Vol.29 No.6

Phosphoinositide turnover in response to platelet-derived growth factor, epidermal growth factor, and bradykinin was evaluated in NIH 3T3 cells. Platelet-derived growth factor and bradykinin induced a significant increase in incorporation of $^{32}P$ into phosphatidylinositol (PI), phosphatidylinositol 4-monophosphate (PIP), and phosphatidylinositol 4.5-bisphosphate ($PIP_2$) in serum-starved NIH 3T3 cells. However, epidermal growth factor increased incorporation of $^{32}P$ into these phosphoinositides by only a small amount. Stimulation with platelet-derived growth factor, not bradykinin, caused a rapid elevation of PI and PIP kinase activities that were maximally activated within 10 min. The maximal levels of their elevation in cells with plateletderived growth factor stimulation were 3.2-fold for PI kinase, and 2.1-fold for PIP kinase. Short term pretreatment of NIH 3T3 cells with phorbol 12-myristate 13-acetate, activator of protein kinase C. caused an approximately 60% decrease in platelet-derived growth factor-induced PI kinase activities, indicating the feedback regulation of phosphoinositide turnover by protein kinase C. These results suggest that although the enhancement of phosphoinositide turnover is a rapidly occurring response in platelet-derived growth factor- or bradykinin-stimulated NIH 3T3 cells, phosphoinositide kinases may be associated with initial signal transduction pathway relevant to platelet-derived growth factor but not to bradykinin.

Effects of Pain-Inducing Agents on Voltage Dependent Calcium Currents in the Trigeminal Ganglion

Park, Kyungpyo,Kim, Seong-Ah,Kim, Youn Bae,Kim, Joong-Soo,Lee, Jong-Heun Korean Academy of Oral Biology and the UCLA Dental 2001 International Journal of Oral Biology Vol.26 No.1

Trigeminal ganglion (TG) neurons are primary sensory neurons involved in the transmission of orofacial sensation, including pain. It is known that intracellular free calcium concentration plays a critical role in neurotransmitter release. In the present study, we examined the effects of pain-inducing agents, bradykinin and histamine, on voltage dependent Ca^2+ current (I_Ca) and compared these with the effects of the analgesic agent, eugenol. I_Ca was measured using a whole-cell patch clamp method. Bradykinin (10μM) inhibited I_Ca in TG neurons. Maximum inward I_Ca was reduced by 26% by bradykinin compared with the controls (n=13, p<0.05). Decreased I_Ca induced by bradykinin did not recover to the prestimulus level after washing out bradykinin. In contrast, histamine (100μM) increased I_Ca by 25% compared with controls (n=7,p<0.05) in an irreversible manner. Eugenol (250μM), commonly used to produce analgesia in dental clinics, decreased I_Ca to 22.6% (n=9,p<0.05) and this effect on the I_Ca was reversible. although the effect of eugenol on the I_Ca was consistent, the effects of the two pain-inducing agents, bradykinin and histamine, were different. Our results suggest that the modulation of I_Ca in TG neurons is not a decisive factor underlying the pain mechanism evoked by pain-inducing agents.