Effect of 6-Hydroxydopamine (6-OHDA) on the Expression of Testicular Steroidogenic Genes in Adult Rats

허현진,안련섭,이성호 한국발생생물학회 2010 발생과 생식 Vol.14 No.3

A neurotoxin, 6-hydroxydopamine (6-OHDA) has been widely used to create animal model for Parkinson's disease (PD). The present study was undertaken to examine whether depletion of brain dopamine (DA) stores with 6-OHDA can make alteration in the activities of the testicular steroidogenesis in adult rats. Young adult male rats (3 months old) were received a single dose of 6-OHDA (200 ㎍ in 10 ㎕/animal) by intracerebroventricular (icv) injection, and sacrificed after two weeks. The mRNA levels of steroidogenesis-related enzymes were measured by qRT-PCRs. Serum testosterone levels were measured by radioimmunoassay. Single icv infusion of 6-OHDA significantly decreased the mRNA levels of CYP11A1 (control:6-OHDA group=1:0.68±0.14 AU, p<0.05), CYP17 (control:6-OHDA group=1:0.72±0.13 AU, p<0.05). There were no changes in the mRNA levels of 3β-HSD (control:6-OHDA group=1:0.84±0.08 AU) and 17β-HSD (control: 6-OHDA group=1:0.63±0.20 AU), though the levels tended to be decreased in the 6-OHDA treated group. Administration of 6-OHDA decreased significantly the mRNA level of StAR when compared to the level of saline-injected control animals (control:6-OHDA group=1:0.72±0.08 AU, p<0.05). Treatment with single dose of 6-OHDA remarkably lowered serum testosterone levels compared to the levels of control group (control:6-OHDA group=0.72±0.24:0.13±0.03 ng/㎖, p<0.05). Taken together with our previous study, the present study demonstrated that the activities of hypothalamus-pituitary-testis hormonal axis could be negatively affected by blockade of brain DA biosynthesis, and suggested the reduced reproductive potential might be resulted in the animals. More precise information on the testicular steroidogenic activities in PD patients and PD-like animals should be required prior to the generalization of the sex steroid hormone therapy to meet the highest standards for safety and efficacy.

플라보노이드 luteolin이 세포자멸사 관련 단백질 활성 억제를 통해 6-hydroxydopamine 세포독성에 미치는 억제효과

이선화,강진호,한정호,김두응,이정수 대한신경과학회 2012 대한신경과학회지 Vol.30 No.4

Background: Flavonoid luteolin has been shown to exhibit cell protective effect. However, it is still uncertain whether the effect of luteolin on cellular toxicity of the parkinsonian toxin 6-hydroxydopamine is mediated by apoptosis-related protein activation. Methods: In differentiated PC12 cells exposed to 6-hydroxydopamine in combination with luteolin, we observed the apoptosis-related protein activation, nuclear damage, formation of reactive oxygen species and cell death. Results: 6-Hydroxydopamine caused apoptosis by inducing a decrease in Bid, Bcl-2, Bcl-xL and survivin levels, increase in Bax levels, cytochrome c release and activation of caspases. Treatment with luteolin reduced changes in the apoptosis-related protein levels, formation of reactive oxygen species, nuclear damage and cell death. Conclusions: Luteolin may reduce the 6-hydroxydopamine-induced apoptosis in differentiated PC12 cells by suppressing the activation of the caspase-8- and Bid-dependent pathways and the mitochondria-mediated apoptotic pathway, leading to caspase activation. The preventive effect of luteolin may be associated with its inhibitory effect on the production of reactive oxygen species. Luteolin may attenuate the oxidative stress and mitochondrial dysfunction-induced neuronal cell death take place in Parkinson’s disease. Background: Flavonoid luteolin has been shown to exhibit cell protective effect. However, it is still uncertain whether the effect of luteolin on cellular toxicity of the parkinsonian toxin 6-hydroxydopamine is mediated by apoptosis-related protein activation. Methods: In differentiated PC12 cells exposed to 6-hydroxydopamine in combination with luteolin, we observed the apoptosis-related protein activation, nuclear damage, formation of reactive oxygen species and cell death. Results: 6-Hydroxydopamine caused apoptosis by inducing a decrease in Bid, Bcl-2, Bcl-xL and survivin levels, increase in Bax levels, cytochrome c release and activation of caspases. Treatment with luteolin reduced changes in the apoptosis-related protein levels, formation of reactive oxygen species, nuclear damage and cell death. Conclusions: Luteolin may reduce the 6-hydroxydopamine-induced apoptosis in differentiated PC12 cells by suppressing the activation of the caspase-8- and Bid-dependent pathways and the mitochondria-mediated apoptotic pathway, leading to caspase activation. The preventive effect of luteolin may be associated with its inhibitory effect on the production of reactive oxygen species. Luteolin may attenuate the oxidative stress and mitochondrial dysfunction-induced neuronal cell death take place in Parkinson’s disease.

Progressive Degeneration of Nigrostriatal Dopaminergic Neurons by Intrastriatal Injection of 6-Hydroxydopamine (6-OHDA) : Protective Effect of Melatonin

Kim, Yong-Sik,Joo, Wan S.,Maeng, Sung H. 경희대학교 2001 INTERNATIONAL SYMPOSIUM ON EAST-WEST MEDICINE Vol.2001 No.1

Although the etiology of Parkinson's disease (PD) remains obscure, the excessive generation of free radicals caused by oxidative stress in nigrostriatal dopaminergic pathway gas been considered as the proximal cause, finally leading to neuronal death. For understanding the pathogenetic mechanisms of PD and evaluating antioxidant neuroprotective drugs, 6-OHDA or MPTP(N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) has been frequently used to make a Parkinsonian animal model. Among the various experimental PD models, a delayed and progressive PD model of the nigrostriatal DA pathway reproducing the symptomatic situation of Parkinsonian patients are interesting to develop. From these background we studied the time course and extend of dopaminergic cell death in substantia nigra (SN) and the striatal dopaminergic dysfunction in the intrastriatal 6-OHDA lesion of rats. 6-OHDA (20ug) was injected unilaterally into the right striatum to induce the nigrostriatal degeneration. Two weeks after 6-OHDA injection, apomorphine-induced contralateral rotational behavior was monitored. The next day animals were sacrificed and brains were removed, processed for the immunohistochemistry and biochemistry. In 6-OHDA-lesioned rats, time-dependent reduction in tyrosine hydroxylase (TH) enzyme activity and dopamine levels was observed in lesioned striatum from 4 days after 6-OHDA injection. In addition, apomorphine produced significant rotational asymmetry 2 weeks after lesioning. Analysis by TH immunocytochemistry revealed the loss of cell bodies in the SN and absence of terminals in the dorsolateral striatum ipsilaterally. One to four weeks after lesioing, a progresive, time-dependent decrease in dopamine transporter (DAT) and vesicular monoamine transporter-2 (VMAT-2) immunoreactivity was also shown in the ipsilateral striatum and SN. We conclude that the injection of 6-OHDA into the terminal field of nigral dopaminergic neurons causes a progressive degeneration of dopamine neurons in SN, starting at one weeks after 6-OHDA injection and continuing over four weeks. Next, we tried to determine whether melatonin, which is a well-known neurohormone and have an antioxidant action, would protect the neuronal damages induced by intrastriatal 6-OHDA injection. Melatonin (3and 10 mg/ kg dissolved in 5% ethanol:saline) or vehicle was administrated intraperitoneally 1 hour prior to 6-OHDA lesion. Following a unilateral injection of 6-OHDA, animals received melatonin treatment, immediately, then every hour for two hours. The next day animals continuously received melatonin once a day for three days.When melatonin was treated in lesioned rats, melatonin significantly reduced the rotational behavior following apomorphine challenge 2 weeks after 6-OHDA lesioning. In parallel, partial, but statistically significant recovery of striatal dopaminergic function, including TH enzyme activity and DA levels also occurred following melatonin treatment. Similar to motor recovery, melatonin treatment resulted in the survival of dopaminergic neurons in SN and restoration of TH immuoreactive terminals in the dorsolateral striatum. Furthermore, melatonin treatment almost completely restored malondialdehyde (lipidperoxidation product) levels to normal, suggesting the in vivo action of melatonin as an antioxidant. These behavioral, histochemical and biochemical results may indicate the neuroprotective actions of melatonin against 6-OHDA- induced toxicity in vivo and suggest the potential pharmaclolgical role in the treatment of oxidative stress-induced neurodegenerative diseases. Also this 6-OHDA-induced PD model may be highly useful in investigating the therapeutic approaches on PD regarding neuronal protection as well as regeneration and metabolic changes of the DA pathway. Key words : 6-hydroxydopamine, Parkinson's disease, Melatonin, Oxidative stress, malondialdehyde, Tyrosine hydroxylase, Dopamine, Striatum.

트레드밀 운동이 6-OHDA로 유발된 파긴슨병 흰쥐 흑색질 내 염증지표에 미치는 영향

조한삼 ( Han Sam Cho ),신말순 ( Mal Soon Shin ),김창주 ( Chang Ju Kim ),백성수 ( Seung Soo Baek ) 한국운동생리학회(구-한국운동과학회) 2012 운동과학 Vol.21 No.1

본 연구에서는 트레드밀 운동이 파킨슨병 쥐의 혹색질에서 염증반응에 미치는 영향을 실험하였다. 6주령의 수컷 쥐를 사용하였으며, 파킨슨병은 뇌정위 수술(stereotaxic surgery) 후 6-hydroxydopamine(6-OHDA)를 주입하여 유발하였다. 각각 가성수술군(sham-operation group; n=10). 가정수술 운동군(sham-operation and exercise group; n=10), 파킨슨병군(6-OHDA-injection group; n=10), 파킨슨병 운동군(6-DHDA-injection and exercise group; n= 10)으로 나누어 파킨슨병 유발 4주후부터 운동군은 하루 30분 총 14일간 트레드밀 달리기를 하였다. 흑색질의 염증반응은 cyclooxygenase-2(COX-2) 및 inducible nitric oxide synthase (iNCG) 면역조직화학법으로 분석하였다. 흑색질의 COX-2 양성세포수는 가정수술군이 54.19±3.68. 운동군이 42.94±3.0l, 파킨슨병 유발군이 98.22±4.73. 파킨슨병 유발 운동군이 61.78±3.86을 나타내었으며, iNOS 양성세포수는 가성수술군이 57.93±3.38. 운동군이 51.64±5.l7. 파킨슨병 유발군이 126.56±7.04, 파킨슨병 유발 운동군이 92.69±8.28을 나타내었다. 본 실험을 통하여 6-OHDA 처치로 인해 염증반응이 증가하였으며, 트레드밀 운동이 COX-2 및 iNOS의 발현을 유의하게 감소시켜, 운동이 파킨슨병으로 인한 뇌신경세포의 염증반응을 완화시키는 긍정적인 효과를 보이는 것을 알 수 있었다. In the present study, we investigated the effects of treadmill exercise on inflammatory reaction in the substantia nigra of the Parkinson`s rats. Parkinson`s disease was made by injection of 6-hydroxyckJpamine (6-OHDA) into the striatum using 6-week-old male rats. The rats were divided into 4 groups: sham-operation group (n=10), sham-operation and exercise group (n=10), 6-OHDA-injection group (n=10), and 6-OHDA-injection and exercise group (n=10). The animals in the exercise groups were put on the treadmill to run for 30 minutes once a day for 14 consecutive days, starting 4 weeks after 6-OHDA injection. For the detection of inflammatory reaction in the substantia nigra immunohistochemistry for cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNCG) was performed The number of CQX-2-positive cells in substantia nigra was 54.19±3.68 in the sham-operation group, 42.94±3.0l in the sham-operation and exercise group, 98.22±4.73 in the 6-OHDA-injection group, and 61.78±3.86 in the 6-OHDA-injection and exercise group. The number of iNOS-positive cells in sull3tantia nigra was 57.93±3.38 in the sham-operation group, 51.64±5.17 in the sham-operation and exercise group, 126.56±7.04 in the 6-OHDA-injection group, and 92.69±8.28 in the 6-OHDA-injection and exercise group. In the present results, 6-OHDA injection increased inflammatory reaction in the substantia nigra, and treadmill exercise alleviated the expression of COX-2 and iNOS in the sull3tantia nigra. These results suggest that treadmill exercise may ameliorate inflammatory reaction in the Parkinson`s disease patients.

6-OHDA로 유도된 파킨슨 증후군의 동물 모델에서 도파민성 신경세포의 손실과 미세아교세포 활성 사이의 관계

이진숙(Jin-Suk Lee),이지용(Ji-Yong Lee),조원길(Won-Gil Cho),양영철(Young-Chul Yang),조병필(Byung-Pil Cho) 대한체질인류학회 2013 해부·생물인류학 (Anat Biol Anthropol) Vol.26 No.1

본 연구는 6-OHDA로 유도된 파킨슨증후군 동물 모델에서 활성화된 미세아교세포의 형태학적 특징 및 면역학적 표현형의 변화와 도파민성 신경세포와의 병리역학적 관계를 확인해 보고자 하였다. 오른쪽 안쪽앞뇌다발에 6-OHDA을 주입하여 흑색질 치밀부에 있는 도파민성 신경세포의 변성을 유도하였다. 동물모델을 6-OHDA 주입 후 1주, 2주, 4주 그리고 8주에 각각 희생시켰다. 활성화된 미세아교세포의 기능적 활성의 변화를 확인하기 위하여 미세아교세포의 포식작용 표지자인 ED1과 쥐 활성미세아교세포 특이적 표지자인 OX6을 사용하여 면역조직화학법 염색을 수행하였다. 활성화된 미세아교세포는 6-OHDA 주입 후 1주에 흑색질 치밀부에 있는 변성된 세포돌기 그리고 도파민성 신경세포의 세포체 등에 선택적으로 부착되어 있는 것을 확인할 수 있었고 이어서 흑색질 치밀부의 도파민성 신경세포와 신경섬유의 뚜렷한 손실을 나타내었다. 포식단계의 활성화된 미세아교세포는 6-OHDA 주입 후 2주에 가장 뚜렷하게 나타났고 시간이 지남에 따라 점진적으로 줄어 들었지만 6-OHDA 주입 후 8주까지 남아 있는 것을 확인하였다. 또한 이는 흑색질 치밀부에 있는 도파민성 신경세포수 계측의 결과와 일치함을 보였다. 이와 같은 결과는 미세아교세포의 활성은 도파민성 신경세포의 변성과정에 영향을 주어 도파민성 신경세포의 손실을 촉진하며 지속적으로 신경세포사를 이끈다는 것을 시사한다. This study assessed the dynamics of morphological and immunophenotypic properties of activated microglia in a 6-hydroxydopamine (6-OHDA) induced Parkinsonian animal model. Neurodegeneration in the substantia nigra pars compacta (SNc) was induced by unilateral injection of 6-OHDA into the medial forebrain bundle. Parkinsonian animal model were sacrificed at 1, 2, 4 and 8 weeks after 6-OHDA injection. Changes in the functional activity of activated microglia were identified using different monoclonal antibodies: OX6 for major histocompatibility complex (MHC) class II, ED1 for phagocytic activity. Phagocytic microglia, characterized by ED1- or OX6-immunoreactivity, appeared in the SNc at 1 week after 6-OHDA injection, activated microglia selectively adhered to degenerating axons, dendrites and dopaminergic neuron somas in the SNc. This was followed by significant loss of these fibers and nigral dopaminergic neurons. Activation of microglia into phagocytic stage was most pronounced at 2 week after 6-OHDA injection and gradually subsided, but phagocytic microglia persisted until 8 weeks after 6-OHDA injection. Taken together, our results indicate that activated microglia is lead to persistently neuron cell death and promotes loss of dopaminergic neuron by degeneration of the dopaminergic neurons.

Protective effects of 6,7,4¢-trihydroxyisofl avone, a major metabolite of daidzein, on 6-hydroxydopamineinduced neuronal cell death in SH-SY5Y human neuroblastoma cells

Yong‑Hyun Ko,Seung‑Hwan Kwon,Seon‑Kyung Kim,이보람,Kwang‑Hyun Hur,Young‑Jung Kim,Seong‑Eon Kim,Seok‑Yong Lee,Choon‑Gon Jang 대한약학회 2019 Archives of Pharmacal Research Vol.42 No.12

Daidzein, one of the important isoflavones,is extensively metabolized in the human body followingconsumption. In particular, 6,7,4′-trihydroxyisoflavone(THIF), a major metabolite of daidzein, has been the focusof recent investigations due to its various health benefits,such as anti-cancer and anti-obesity effects. However, theprotective effects of 6,7,4′-THIF have not yet been studied inmodels of Parkinson’s disease (PD). Therefore, the presentstudy aimed to investigate the protective activity of 6,7,4′-THIF on 6-hydroxydopamine (OHDA)-induced neurotoxicityin SH-SY5Y human neuroblastoma cells. Pretreatmentof SH-SY5Y cells with 6,7,4′-THIF significantly inhibited6-OHDA-induced neuronal cell death, lactate dehydrogenaserelease, and reactive oxygen species production. In addition,6,7,4′-THIF significantly attenuated reductions in 6-OHDAinducedsuperoxide dismutase activity and glutathione content. Moreover, 6,7,4′-THIF attenuated alterations in Baxand Bcl-2 expression and caspase-3 activity in 6-OHDAinducedSH-SY5Y cells. Furthermore, 6,7,4′-THIF significantlyreduced 6-OHDA-induced phosphorylation of c-JunN-terminal kinase, p38 mitogen-activated protein kinase,and extracellular signal-regulated kinase 1/2. Additionally,6,7,4′-THIF effectively prevented 6-OHDA-induced loss oftyrosine hydroxylase. Taken together, these results suggestthat 6,7,4′-THIF, a major metabolite of daidzein, may be anattractive option for treating and/or preventing neurodegenerativedisorders such as PD.

6-Hydroxydopamine으로 유도된 질소적 세포 사멸에 대한 고려홍삼 추출물의 보호효과

이찬(Chan Lee),장정희(JungHee Jang),박규환(GyuHwan Park) 대한약학회 2016 약학회지 Vol.60 No.2

Parkinson’s disease (PD) is one of the representative neurodegenerative movement disorders with the selective loss of dopaminergic neurons in the substantia nigra. 6-Hydroxydopamine (6-OHDA) is widely used as an experimental model system to mimic PD and has been reported to cause neuronal cell death via oxidative and/or nitrosative stress. Therefore, daily intake of dietary or medicinal plants which fortifies cellular antioxidant capacity can exert neuroprotective effects in PD. In the present study, we have investigated the protective effect of Korean red ginseng (KRG) against 6-OHDAinduced nitrosative death in C6 glioma cells. Treatment of C6 cells with 6-OHDA decreased cell viability and increased expression of inducible nitric oxide synthase, production of nitric oxide as well as peroxynitrite, and formation of nitrotyrosine. 6-OHDA led to apoptotic cell death as determined by decreased Bcl-2/Bax, phosphorylation of JNK, activation of caspase-3, and cleavage of PARP. Conversely, pretreatment of C6 cells with KRG attenuated 6-ODHA-induced cytotoxicity, apoptosis, and nitrosative damages. To further elucidate the molecular mechanism of KRG protection against 6-OHDAinduced nitrosative cell death, we have focused on the cellular self-defense molecules against exogenous noxious stimuli. KRG treatment up-regulated heme oxygenase-1 (HO-1), a key antioxidant enzyme essential for cellular defense against oxidative and/or nitrosative stress via activation of Nrf2. Taken together, these findings suggest KRG may have preventive and/ or therapeutic potentials for the management of PD.

Effect of 6-Hydroxydopamine (6-OHDA) on the Expression of Testicular Steroidogenic Genes in Adult Rats

Heo Hyun-Jin,Ahn Ryun-Sup,Lee Sung-Ho 한국발생생물학회 2010 발생과 생식 Vol.14 No.3

A neurotoxin, 6-hydroxydopamine (6-OHDA) has been widely used to create animal model for Parkinson's disease (PD). The present study was undertaken to examine whether depletion of brain dopamine (DA) stores with 6-OHDA can make alteration in the activities of the testicular steroidogenesis in adult rats. Young adult male rats (3 months old) were received a single dose of 6-OHDA (200 in 10 /animal) by intracerebroventricular (icv) injection, and sacrificed after two weeks. The mRNA levels of steroidogenesis-related enzymes were measured by qRT-PCRs. Serum testosterone levels were measured by radioimmunoassay. Single icv infusion of 6-OHDA significantly decreased the mRNA levels of CYP11A1 (control:6-OHDA group= AU, p<0.05), CYP17 (control:6-OHDA group= AU, p<0.05). There were no changes in the mRNA levels of -HSD (control:6-OHDA group= AU) and -HSD (control: 6-OHDA group= AU), though the levels tended to be decreased in the 6-OHDA treated group. Administration of 6-OHDA decreased significantly the mRNA level of StAR when compared to the level of saline-injected control animals (control:6-OHDA group= AU, p<0.05). Treatment with single dose of 6-OHDA remarkably lowered serum testosterone levels compared to the levels of control group (control:6-OHDA group=, p<0.05). Taken together with our previous study, the present study demonstrated that the activities of hypothalamus-pituitary-testis hormonal axis could be negatively affected by blockade of brain DA biosynthesis, and suggested the reduced reproductive potential might be resulted in the animals. More precise information on the testicular steroidogenic activities in PD patients and PD-like animals should be required prior to the generalization of the sex steroid hormone therapy to meet the highest standards for safety and efficacy.

6-Hydroxydopamine 유발 SH-SY5Y 세포주 손상에 대한 resveratrol의 신경보호 효과

장건천,김형춘,위명복,Chang, Geon-Cheon,Kim, Hyoung-Chun,Wie, Myung-Bok 대한수의학회 2014 大韓獸醫學會誌 Vol.54 No.1

Parkinson's disease is known to exhibit progressive degeneration of the dopaminergic neurons in the substantia nigra via inhibition of glutathione metabolism. It is well known that 6-Hydroxydopamine (6-OHDA) induces Parkinson's disease-like symptoms, while resveratrol (3,5,4'-trihydroxystilbene) has been shown to have anti-inflammatory and antioxidant effects. In the present study, we investigated the neuroprotective effects of resveratrol, a phytoalexin found in grapes and various plants, on 6-OHDA-induced cell damage to the SH-SY5Y human neuroblastoma cell line. Resveratrol (5 and 10 ${\mu}M$) inhibited 6-OHDA (60 ${\mu}M$)-induced cytotoxicity in SH-SY5Y cells and induced a reduction of the number of apoptotic nuclei caused by 6-OHDA treatment. Additionally, the total apoptotic rate of cells treated with both resveratrol (10 ${\mu}M$) and 6-OHDA (60 ${\mu}M$) was less than that of 6-OHDA treated cells. Resveratrol also dose-dependently (1, 5 and 10 ${\mu}M$) scavenged reactive oxygen species (ROS) induced by 6-OHDA in SH-SY5Y cells and prevented depletion of glutathione in response to the 6-OHDA-induced cytotoxicity in the glutathione assay. Overall, these results indicate that resveratrol exerts a neuroprotective effect against 6-OHDA-induced cytotoxicity of SH-SY5Y cells by scavenging ROS and preserving glutathione.

Berberine이 백서의 6-hydroxydopamine-유도 파킨슨병 모델에 미치는 영향

권익현,최현숙,신건성,황방연,이명구 한국생약학회 2009 생약학회지 Vol.40 No.4

Many isoquinoline alkaloids including berberine lower dopamine content by reducing tyrosine hydroxylase (TH) activity and aggravate L-DOPA-induced cytotoxicity in PC12 cells. In this study, the effects of berberine on 6-hydroxydopamine (6-OHDA)-induced cytotoxicity in PC12 cells and on unilateral 6-OHDA-lesioned rat models were investigated. Berberine at 10-30 µM did not affect cell viability in PC12 cells. However, berberine at concentrations higher than 50 µM caused cytotoxicity at 24 h. Berberine (10-50 µM) also enhanced 6-OHDA (10-50 µM)-induced cytotoxicity at 24 h compared to 6-OHDA alone with an apoptotic process. In addition, treatment with berberine (5 and 30 mg/kg, i.p.) for three weeks showed a dopaminergic cell loss in substantia nigra of 6-OHDA-lesioned rats: 30 mg/kg berberine was more intensive cytotoxic. The levels of dopamine were also decreased by berberine (5 and 30 mg/kg) in the ipsilateral substantia nigra of 6-OHDA-lesioned rats. These results suggest that berberine aggravated 6-OHDA-induced cytotoxicity in PC12 cells and treatment with berberine (5 and 30 mg/kg) in 6-OHDA-lesioned rats also enhanced the degeneration of dopaminergic cell death and the decrease in dopamine levels in substantia nigra. Therefore, the long-term L-DOPA therapeutic patients with isoquinoline compounds including berberine may need to be checked for the adverse symptoms.