트레드밀 운동이 Streptozotocin 유발 당뇨쥐 해마에서 항산화 효소 수준 증가 및 Apoptosis 억제에 미치는 효과

채창훈(ChangHunChae),김현태(HyunTaeKim) 한국체육학회 2011 한국체육학회지 Vol.50 No.6

본 연구는 규칙적인 운동이 streptozotocin(STZ) 유도 당뇨 쥐 해마에서 Mn-SOD, Cu, Zn-SOD의 단백질 수준증가와 apoptosis 억제 효과를 살펴보았다. 연구 대상은 생후 8주령된(225±5.2g) SD계 흰쥐 40마리를 정상통제군(NCG; n=10), 정상운동군(NEG, n=10), 당뇨통제군(DCG, n=10) 그리고 당뇨운동군(DEG, n=10)으로 각각 무선배정하였다. DCG와 DEG 쥐들에게는 점진적 과부하의 원리를 적용시켜 주 5일(30분/일), 6주간 규칙적인 트레드밀 운동을 실시하였다. 그 결과 고혈당을 동반하는 당뇨병시에는 Mn-SOD, Cu, Zn-SOD의 단백질 수준이 저하되고 apoptosis가 촉진되는 것으로 확인되었다. 반면, 규칙적인 운동을 실시한 결과 해마의 Mn-SOD, Cu, Zn-SOD의 단백질 발현이 증가되는 반면, apoptosis가 억제되는 것으로 밝혀졌다. 이러한 결과는 당뇨병시 발생되는 항산화 효소의 활성저하와 세포사멸 현상을 규칙적인 운동을 통해 억제함으로써 운동이 당뇨병시 해마의 기능저하를 예방하는데 긍정적인 기여를 할 것으로 사료된다. We investigated the effects of regular treadmill exercise for 6 weeks on Mn-SDO, Cu, Zn-SOD protein levels and apoptosis in the streptozotocin-induced diabetic rat hippocampus. Forty male Sprague-Dawley rats were divided into four groups: (1) NCG (n=10); (2) NEG (n=10); (3) DCG (n=10); (4) DEG (n=10). Diabetes was induced by injecting streptozotocin (STZ; 55mg/kg dissolved in 0.05M citrate buffer, p.H 4.5, i.p.) into rats. Rats were subjected to treadmill exercise for 5 days a week over 6 weeks, and the speed of the treadmill was gradually increased. Mn-SDO, Cu,Zn-SOD protein levels significantly decreased DCG. Also, Apoptosis by Tunel assay showed a significant induction in apoptosis increased in the DCG. Whereas decreased apoptosis and significant increases in Mn-SDO, Cu,Zn-SOD expression levels significantly was decreased and apoptosis significantly is increased in the hippocampus of STZ-diabetic rats(DCG), but treadmill exercise(DEG) significantly increased expression of Mn-SOD and Cu,Zn-SOD, and decreased apoptosis compare to DCG. These results show that exercise may improves function of the hippocampus via increase Mn-SDO and Cu,Zn-SOD expression levels and suppression of apoptosis in the the hippocampus of STZ-induced diabetic rats.

트레드밀 운동이 당뇨 쥐 soleus에서 Mn-SOD, Cu,Zn-SOD, caspase-3 단백질 수준 및 혈당조절에 미치는 영향

채창훈(ChangHunChae),김현태(HyunTaeKim) 한국체육학회 2009 한국체육학회지 Vol.48 No.6

본 연구는 STZ 투여에 의해 당뇨 유발된 흰쥐를 대상으로 6주간의 규칙적인 운동이 골격근 selous의 antioxidant enzyme 및 GULT-4, blood glucose와 caspase-3의 protein 발현에 미치는 효과를 검증하고자, 흰쥐 20마리를 각 집단간 5마리씩 배정하여 총 4개 집단(정상통제군: NCG, 정상운동군: NEG, 당뇨통제군: DCG, 당뇨운동군: DEG)으로 무선배정 하여 6주간 각 집단별 처치를 실시하였다. 그 결과 운동군의 Mn-SOD와 Cu,Zn-SOD, GLUT-4와 caspase-3 단백질 발현이 통계적으로 유의한 차이(각각 p<0.001)를 보였으며, 당뇨집단에서도 당뇨통제군에 비해 당뇨운동군의 혈당 및 Mn-SOD와 Cu,Zn-SOD, GLUT-4 단백질 발현이 통계적으로 유의한 차이(p<.001)를 나타냈다. 그러나 당뇨군의 caspase-3 단백질 발현은 통계적으로 유의한 차이가 나타나지 않았다. 이상을 종합해 보면 정상적인 상태에서 뿐 만 아니라 당뇨병과 같은 병리적 조건에서도 6주간의 규칙적인 운동은 항산화 방어계를 강화시킴으로서 혈당조절 및 GLUT-4의 단백질 발현에 효과가 있는 것으로 확인되었으나, caspase-3의 저하에는 영향을 미치지 못하는 것으로 확인되었다. The purpose of this study was to investigate the effects of treadmill exercise done regularly for 6 weeks on blood glucose, GLUT-4 and Mn-SOD, Cu,Zn-SOD protein levels in soleus of STZ-induced diabetic rats. Twenty male SD rats were divided into the following 4 groups; (1) normal control group(NCG: n=5); (2) normal exercise group(NEG; N=5); (3) diabetes control group(DCG: n=5); and diabetes exercise group(DEG: n=5). Diabetes was induced by injecting WTZ(55mg/kg/b.w.). Rats were subjected to treadmill exercise for 5 days a week for 6 weeks. Blood glucose level in DCG and DEG was significantly increased than that in NCG and NEG. Especially, blood glucose in DEG was significantly decreased than that in DEG. Mn-SOD, and Cu,Zn,-SOD protein levels significanlty increased in NEG, furthermore, Mn-SOD and Cu,Zn-SOD protein levels in DEG were significantly increased than that in DCG. GLUT-4 protein level significantly increased in NEG and DEG. Caspse-3 protein level significantly decreased in NEG. These results show that treadmill exercise in considered to improve blood glucose and GLUT-4 protein level by increased antioxidant emzymes protein levels in soleus of STZ-induced diabetic rats.

2시간 및 6시간 허혈시킨 흰쥐의 가자미근에서 재관류 시간경과에 따라 나타나는 SOD 활성변동에 대한 연구

황규성,안동춘,서윤경,백두진,정호삼,노원재,이문구 한양대학교 의과대학 2000 한양의대 학술지 Vol.20 No.1

It has been well known that ischemia reperfusion injury to skeletal muscle following an acute arterial occulusion causes significant morbidity and mortality. The skeletal muscle, which contains high energy phosphate compounds, has ischemic tolerance. During the ischemia, the ATP is catalyzed to hypoxanthine anaerobically and hypoxanthine dehydrogenase is converted to xanthine oxidase. During reperfusion, the hypoxanthine is catalyzed to xanthine by xanthine oxidase undr O_2 presence and that results in production of cytotoxic oxygen free radicals. So, the damage of skeletal muscle occrus during reperfusion. The permeability of the capillary, one of the major phenomenon in injured muscle, is increased by reactive oxygen released from the neutrophils and the arachidonate metabolites, which are catalyzed from the lipid component of the membrane structures by lipooxygenase during reperfusion. The reversible changes in the muscle occur after 2hours of ischemia and reperfusion and recover within 24 hours. The irreversible damage, which is necrosis of muscle, occurs after 6 hours of ischemia and reperfusion. The reactivity of superoxide dismutase(SOD), one of the antioxidant enzymes is increased against the formation of superoxide radical during the reperfusion. The SOD metabolizes the superoxide radical to H_2O_2 and O_2. There are two types of SOD in the cell. The one of them is Cu,Zn-SOD in the cytoplasm and the other is Mn-SOD in the mitochondria. The author performes the present study to investigate the changes of SOD immunoreactivities in the soleus muscle of the rat after 2 hours or 6 hours of ischemia and timely reperfusion. A total of 88 healthy Sprague-Dawley rats weighing from 200 gm to 250 gm were used as experimental animals. Under urethane(3.0g/kg., i.p.) anesthesia, lower abdominal incision was made and left common iliac artery were ligated by using vascular clamp for 2 hours. Soleus muscles were obtained at 0 hour, 1 hour, 2 hours, 24 hours 48hours after the removal of vascular clamp. The specimens were sectioned in 14㎛ thickness with cryostat. The immunoreactivities of SOD by use of anti human Cu,Zn- and Mn-SOD antibodies were observed. The results were as follows. 1. The immunoreactivities of SOD around sarcolemma were stronger than those in the sarcoplasm. 2. The immunoreactivities of SOD in the soleus muscles decreased a little after 2 hours of ischemia and increased markedly after 6 hours of ischemia. 3. The immunoreactivities of Cu,Zn-SOD in the soleus muscle increased after 2 hours or 6 hours of ischemia and timely reperfusion. The immunoreactivities of Cu,Zn-SOD in the 6 hours of reperfused group were highest after 2 hours of ischemia. Those in the 2 hours of reperfused group were highest after 6 hours of ischemia. At 48 hours of reperfusion, immunoreactivities of Cu,Zn-SOD recovered after 2 hours of ischemia and those kept higher reactivities after 6 hours of ischemia. 4. The immunoreactivities of Mn-SOD in the soleus muscles increased after 2 hours or 6 hours of ischemia and timely reperfusion. At 48 hours of reperfusion, immunoreactivities of Mn-SOD recovered after 2 hours of ischemia and those were increased markedly after 6 hours of ischemia. These results suggest that the immunoreactivities of SOD increased after 2 hours or 6 hours of ischemia and timely reperfusion. At 48 hours of reperfusion the immunoreactivities of SOD recover after 2 hours of ischemia and keep higher reactivities after 6 hours of ischemia.

Hepatic Expression of Cu/Zn-Superoxide Dismutase Transcripts in Response to Acute Metal Exposure and Heat Stress in Hemibarbus mylodon (Teleostei: Cypriniformes)

Cho, Young-Sun,Bang, In-Chul,Lee, Il-Ro,Nam, Yoon-Kwon The Korean Society of Fisheries and Aquatic Scienc 2009 Fisheries and Aquatic Sciences Vol.12 No.3

Hemibarbus mylodon (Cypriniformes) is an endemic freshwater fish species in the Korean peninsula, for which urgent conservation efforts are needed. To understand their stress responses in relation to metal toxicity and thermal elevation, we performed a real-time RT-PCR-based expression assay of hepatic copper/zinc-superoxide dismutase (Cu/Zn-SOD), a key antioxidant enzyme, in response to experimental heavy metal exposure or heat treatment. The transcription of hepatic Cu/Zn-SOD was differentially modulated by acute exposure to Cu, cadmium (Cd), or Zn. Exposure to each metal at $5{\mu}M$ for 24 h revealed that Cu stimulated the mRNA expression of Cu/Zn-SOD to a greater extent than the other two heavy metals. The elevation in Cu/Zn-SOD transcripts in response to Cu exposure was dose-dependent (0.5 to $5{\mu}M$). Time course analysis of Cu/Zn-SOD expression in response to Cd exposure ($5{\mu}M$) revealed a transient pattern up to day 7. Exposure to thermal stress (an increase from 22 to $30^{\circ}C$ at a rate of $1^{\circ}C/h$ followed by $30^{\circ}C$ for 18 h) did not significantly alter SOD transcription, although heat shock protein 90 kDa (HSP90) transcription was positively correlated with an increase in temperature.

Transduction Efficacy of Tat-Cu,Zn-Superoxide Dismutase Is Enhanced by Copper Ion Recovery of the Fusion Protein

음원식,In Soon Choung,A Yeon Kim,Yong Je Lee,Jung Hoon Kang,Jinseu Park,Kil Soo Lee,Hyeok Yil Kwon,최수영 한국분자세포생물학회 2002 Molecules and cells Vol.13 No.2

We previously reported that Tat-Cu,Zn-superoxide dismutase (Tat-SOD) can be directly transduced into mammalian cells across the lipid membrane barrier. To enhance the therapeutic potential of Tat-SOD for the treatment of various disorders that are related to this antioxidant enzyme, the transduction efficacy of Tat-SOD should be heightened. Therefore, we investigated whether copper ion recovery of the fusion protein could enhance the transduction potential of Tat- SOD in cultured HeLa cells. The results showed that the transduction potential of Tat-SOD was markedly enhanced by copper ions, and moderately increased by zinc ions. Compared with Tat-SOD, the Tat-SOD that recovered the copper ion (CR-Tat-SOD) achieved a significant increase in intracellular concentration and enzymatic activity. Therefore, CR-Tat-SOD was transduced into HeLa cells in a rapid saturation manner, but Tat-SOD was shown in a time-dependent manner. With the higher transduction efficacy of CR-Tat-SOD than that of Tat-SOD, the transduced CR-Tat-SOD significantly increased the viability of HeLa cells that were pretreated with paraquat, an intracellular superoxide anion generator. Although the mechanism of the enhanced transduction of Tat-SOD by copper ions is still unanswered, these results indicate that copper ions facilitate the transduction of SOD. These then significantly increase the biological effectiveness of this antioxidant enzyme.

트레드밀 지구성 운동이 streptozotocin으로 유발된 당뇨 흰쥐의 뇌에서 PGC-1α, GLUT-1, Tfam 단백질 및 항산화 효소(Cu, Zn-SOD, Mn-SOD)의 발현량에 미치는 영향

Noh-Hwan Park(박노환),Jin Lee(이진),Kook-Hyun Jung(정국현),Bong-Am Choi(최봉암),Hyung-Chae Jang(장형채),Suk-In Lee(이석인),Dong-Soo Lee(이동수),Joon-Yong Cho(조준용) 한국생명과학회 2011 생명과학회지 Vol.21 No.3

이 연구는 지구성 운동이 streptozotocin (STZ)으로 유발된 제1형 당뇨 특징을 가진 쥐 뇌의 글루코스 운반, 미토콘드리아 기능 및 항산화효소 단백질 발현에 미치는 영향을 규명하는데 목적이 있다. 제1형 당뇨 모델 쥐는 50 ㎎/㎏의 streptozotocin을 수컷 Sprague-Dawley (SD) 흰쥐의 복강에 1회 주입하여 생산하였으며 본 실험 시집단은 NON-STZ 집단(n=8), STZ-CON 집단(n=8) 및 STZ-EXE 집단(n=8) 등 3집단으로 구분하여 실시하였다. 트레드밀 지구성 운동은 총 6주, 주 5일, 2주 간격으로 속도를 약 3∼4 m/min으로 점증적으로 증가시켰으며 운동시간은 1주와 3주차에 10분씩 증가시켰다. 분석 결과 혈청 글루코스 수준은 STZ-EXE 집단은 STZ-CON 집단에 비해 현저하게 감소(p<0.05)하였으며 PGC-1α (p<0.001), mtPGC-1α (p<0.001), GLUT-1 (p<0.001), Tfam(p<0.001), Cu,Zn- SOD (p<0.001), Mn-SOD (p<0.01) 경우도 STZ-EXE 집단이 STZ-CON 집단에 비해 현저하게 증가하였다. 이러한 결과는 장기간 지구성 운동이 뇌의 글루코스 이용능력과 관련된 단백질인 GLUT-1과 미토콘드리아 기능 향상과 관련된 단백질인 PGC-1α과 Tfam을 증가시키고 산화적 스트레스의 방어 기전으로서 역할을 수행하는 항산화 효소인 Cu,Zn-SOD와 Mn-SOD를 활성화시키는데 긍정적인 역할을 수행한 것으로 나타났다. The purpose of this study is to identify the effects of exercise training [ET, 10~18 m/min (speed), 20~30 min (exercise duration)/a day for 5 day/wk, 6 wk) on PGC-1α, GLUT-1, Tfam, Cu,Zn-SOD and Mn-SOD proteins in brain of STZ-induced diabetic rats. The male Sprague-Dawley (SD) rats were single-injected intraperitoneally with 50㎎/㎏ of streptozotocin (STZ) to produce STZ-induced diabetic rats. Rats were divided into 3 experimental groups with 8 rats in each group, as follows: (1) non-STZ group (n=8), (2) STZ-CON group (n=8), (3) STZ-EXE group (n=8). The results of this study suggest that i) serum glucose level was significantly reduced in STZ-EXE group compared with STZ-CON group (p<0.05), ii) PGC-1α (p<0.001), mtPGC-1α (p<0.001), GLUT-1 (p<0.001), and mtTfam (p<0.001) proteins in brain of STZ-induced diabetic rats were significantly increased in STZ-EXE group compared with STZ-CON group, iii) Cu,Zn-SOD (p<0.001) and Mn-SOD (p<0.01) proteins in the STZ-induced diabetic rats were significantly increased in STZ-EXE group compared with STZ-CON group. In conclusion, the findings of the present study reveal that treadmill exercise training increases brain GLUT-1 protein level possibly through up-regulation of PGC-1α and Tfam proteins which represent key regulatory components of stimulation of brain mitochondrial biogenesis. In addition, treadmill exercise training may prevent oxidative stress by up-regulation of Cu,Zn-SOD and Mn-SOD proteins in the STZ-induced diabetic rats.

Hepatic Expression of Cu/Zn-Superoxide Dismutase Transcripts in Response to Acute Metal Exposure and Heat Stress in Hemibarbus mylodon (Teleostei: Cypriniformes)

조영선,방인철,이일로,남윤권 한국수산과학회 2009 Fisheries and Aquatic Sciences Vol.12 No.3

Hemibarbus mylodon (Cypriniformes) is an endemic freshwater fish species in the Korean peninsula, for which urgent conservation efforts are needed. To understand their stress responses in relation to metal toxicity and thermal elevation, we performed a real-time RT-PCR-based expression assay of hepatic copper/zinc-superoxide dismutase (Cu/Zn-SOD), a key antioxidant enzyme, in response to experimental heavy metal exposure or heat treatment. The transcription of hepatic Cu/Zn-SOD was differentially modulated by acute exposure to Cu, cadmium (Cd), or Zn. Exposure to each metal at 5 μM for 24 h revealed that Cu stimulated the mRNA expression of Cu/Zn-SOD to a greater extent than the other two heavy metals. The elevation in Cu/Zn-SOD transcripts in response to Cu exposure was dose-dependent (0.5 to 5 μM). Time course analysis of Cu/Zn-SOD expression in response to Cd exposure (5 μM) revealed a transient pattern up to day 7. Exposure to thermal stress (an increase from 22 to 30°C at a rate of 1°C/h followed by 30°C for 18 h) did not significantly alter SOD transcription, although heat shock protein 90 kDa (HSP90) transcription was positively correlated with an increase in temperature.

Modification and Inactivation of Human Cu,Zn-Superoxide Dismutase by Methylglyoxal

Kang, Jung Hoon Korean Society of Molecular Biology 2003 Molecules and cells Vol.15 No.2

흰쥐 귀밑샘에서 자유유리기 제거제가 방사선 조사로 유도된 세포자멸 발현에 미치는 영향

김희진 대한해부학회 1998 Anatomy & Cell Biology Vol.31 No.6

흰쥐 귀밑샘에서 자유유리기 제거제가 방사선 조사로 유도된 세포자멸 발현에 미치는 영향

김희진(Hee-Jin Kim) 대한해부학회 1998 Anatomy & Cell Biology Vol.31 No.6

다핵기관이 생물학적 평형을 유지하기 위해서는 불필요한 세포들의 제거과정이 필수적이며 이는 발생이나 생후 발육 과 정 중 정상조직을 재형성하는 과정에서 나타나는 생리학적 세포 변화의 한 유형인 예정된세포죽음으로 정의 내려진다. 머리와 목 부위의 암을 치료하기 위해 사용하는 치료방사선은 조사후 구강건조증을 일으키며, 이 과정은 방사선에 의한 침샘세포의 인위적인 세포자멸에 의한 것으로 보고되고 있다. 따라서 이 연구는 이온화 방사선에 의한 침샘의 손상을 차단하고 이에 따르는 합병증을 줄이기 위하여 세포자멸을 차단시키는 물질로 알려진 allopurinol, Cu/Zn-superoxide dismutase (Cu/Zn-SOD), 그리고 dimethylthiourea (DMTU)를 방사선 조사 전에 실험동물에 전처치하여 방사선으로 유도되는 세포자멸의 발현을 억제시키고자 하였다. Xanthine oxidase에 의한 초산화물 음이온 생성을 촉진시키는 반응을 억제하는데 효과가 있는 것으로 알려진 allopurinol을 방사선조사 전에 실험동물에 전처치한 결과, 방사선에 의해 유도되는 침샘의 세포자멸 과정을 억제하지는 못하였다. 또한 세포외부에서 발생되는 초산화물을 효과적으로 제거하는 Cu/Zn-SOD와 수산화 자유유리기를 제거하는 효과를 가진 DMTU를 방사선 조사전 실험 동물에 각각 전처치한 결과, 두 가지 약물 모두 방사선에 의한 침샘 세포의 세포자멸 발현을 억제하였으며 (P<0.001), 방사선에 의한 침샘 구조의 손상도 감소되었다. 자유유리기 제거제의 약물 상협효과를 확인하기 위해 방사선 조사전 실험 동물에 allopurinol과 Cu/Zn-SOD, Cu/Zn-SOD와 DMTU를 동시에 전처치한 결과, 방사선에 의해 유도되는 침샘세포의 세포자멸은 자유유리기 제거제를 전처치하지 않고 방사선조사를 시행한 실험군보다는 낮았으나 (P<0.001), 두 가지 약물의 상협효과는 없었다. 이상의 결과를 종합하여 볼 때, 흰쥐 침샘에서 이온화 방사선으로 유도된 세포자멸은 산소 또는 수산화 자유유리기에 의 한 일차적인 세포손상의 결과로 예상되며 자유유리기 제거제의 전처치는 방사선에 의한 세포자멸을 차단할 수 있다는 사실을 확인하였다. 그러나 allopurinol을 전처리하였을 때, 방사선에 의한 세포자멸 양상이 감소되지 않은 것으로 보아 방사선에 의해 생성된 산소 자유 유리기는 xanthine oxidase 과정을 통해 형성된 것이 아님을 알 수 있었다. 따라서 세포자멸에는 여러가지 요인이 작용할 것으로 추측된다. 그러므로 방사선에 의한 초기 세포자멸 과정을 통해 세포의 자가소실과정이 유발되고, 이 과정에서 일어나는 일련의 세포 반응들에 의하여 기능적 손실이 유발될 것으로 예상된다. Apoptosis is a distinct morphological form of programmed cell death during growth, development and some diseases. Also, it is well known that the apoptosis is usually caused by carcinogens, drugs, heat and radiation in mature cells. Especially, the therapeutic radiation of the head and neck region may cause xerostomia after irradiation. Recently, there are many trial reports to explain the radiation cell death to the acute cellular changes as a programmed cell death. The author performed present study to investigate the potential role of reactive oxygen metabolites as a signal for radiation-induced apoptosis. A series of free radical scavengers, allopurinol, superoxide dismutase, dimethylthiourea were pretreated before γ-irradiation for evaluating their ability to block apoptosis in the rat parotid gland in vivo, and the light microscopic results were evaluated and analyzed using the Tdt-mediated dUTPbiotin nick end labeling technique. The results were as follows. 1. Under the light microscope, irradiation induced apoptotic bodies in the parotid gland were revealed the intensively positive reaction on the TUNEL technique, and most apoptotic bodies were confined in the acinar system, there were rare apoptotic changes in the ductal system. 2. In all irradiated experimental groups without antioxidants pretreatment, the number of the apototic bodies was the greatest in post-irradiation 24 hour group, and then decreased as a function of time. 3. As a result on pretreatment of allopurinol that effectively inhibit xanthine oxidase before irradiation, failed to block the irradiation-induced apoptosis in the rat parotid gland. Moreover, in the postirradiation 1 day group, the apoptotic expression and destruction of the acini increased than non-irradiated experimental group. 4. As a result on pretreatment of Cu/Zn-SOD that contributes to extracellularly generated superoxides and dimethylthiourea to scavenge the hydroxyl free radicals before irradiation, two antioxidants inhibit the irradiationinduced apoptosis (P<0.001), salivary acinar structures remained intact. 5. To evaluate the synergistic effects, simultaneous pretreatments of allopurinol and Cu/Zn-SOD, Cu/Zn-SOD and DMTU were conducted. Expression of irradiation-induced apoptosis was lower than the experimental groups which not pretreated any antioxidants (P<0.001), but there was no synergism of two antioxidants. 6. Irradiation-induced apoptotic bodies crowded at the outer cortical layer including lymphatic nodules, and the aspect of apoptotic expression in the lymph node took place earlier than in the salivary acini. Taken all together, it is forecasted that the irradiation-induced apoptosis of the parotid gland is a result of primary cellular damage by oxygen or hydroxyl free radicals. Through this study, the facts that pretreatments of free radical scavengers block the irradiation-induced apoptosis were identified. But, oxygen free radicals were not produced via xanthine oxidase process, whereas, allopurinol that inhibits this reaction influenced the harmful effects on the parotid gland.