KCC2 expression changes in Diazepam-treated neonatal rats with hypoxia-ischaemia brain damage.
Hypoxia-ischaemia brain damage (HIBD) is a major type of perinatal brain injury in newborns. In this study, we investigate the short- and long-term neuroprotective effects of Diazepam on neonatal rats with HIBD and the potential mechanisms underlying its protective effects. Seven-day-old Sprague-Dawley rats were subjected to left carotid artery ligation followed by a 2-h exposure to 8% oxygen and 92% nitrogen. Diazepam was administered immediately via intraperitoneal (i.p.) injection after inducing HIBD at a dose of 10 mg kg(-1)8h(-1) for three consecutive days. Three days after HIBD, rats were decapitated, and the extent of brain injury was evaluated using 2,3,5-triphenyltetrazolium chloride (TTC) staining. Additionally, the expression of Potassium-chloride cotransporter-2 (KCC2) was analysed using real-time PCR, Western blot analysis and immunohistochemistry. Three weeks after HIBD, rats were subjected to the Morris water maze (MWM) test and the locomotor activity test to determine the long-term therapeutic effects of Diazepam. We observed that the volume of infarction in the Diazepam group was significantly less (P<0.01) compared with the HIBD group. We also observed that the learning and memory abilities of the Diazepam rats improved significantly compared with the untreated rats (P<0.05) and that the decrease in KCC2 expression was prevented (P<0.01). Early treatment with Diazepam appears to attenuate HIBD and can efficiently improve the long-term learning and memory capabilities of the animal. A potential mechanism underlying these effects may involve preventing the decrease in KCC2 expression.
School of Pharmaceutical Science, Zhengzhou University, Ke Xue Da Dao 100, Zhengzhou 450001, PR China.
Jun-Yuan
JY
Ma
Medical College of Kaifeng University, Kaifeng 475001, PR China.
Su-Pei
SP
Zhang
School of Pharmaceutical Science, Zhengzhou University, Ke Xue Da Dao 100, Zhengzhou 450001, PR China.
Liu-Bin
LB
Guo
School of Pharmaceutical Science, Zhengzhou University, Ke Xue Da Dao 100, Zhengzhou 450001, PR China.
Yong-Mei
YM
Li
The First Affiliated Hospital of Henan College of Traditional Chinese Medicine, Zhengzhou 450000, PR China; Key Laboratory of Viral Diseases Prevention and Treatment of Traditional Chinese Medicine of Henan Province, Zhengzhou 450000, PR China.
Qiang
Q
Li
School of Pharmaceutical Science, Zhengzhou University, Ke Xue Da Dao 100, Zhengzhou 450001, PR China.
Sai-Qi
SQ
Wang
School of Pharmaceutical Science, Zhengzhou University, Ke Xue Da Dao 100, Zhengzhou 450001, PR China.
Hong-Min
HM
Liu
School of Pharmaceutical Science, Zhengzhou University, Ke Xue Da Dao 100, Zhengzhou 450001, PR China. Electronic address: wangcong@zzu.edu.cn.
Cong
C
Wang
D000818:Animals; D000831:Animals, Newborn; D003975:Diazepam / Q000627:therapeutic use*; D020925:Hypoxia-Ischemia, Brain / Q000188:drug therapy*; D008297:Male; D018782:Maze Learning / Q000187:drug effects; D009043:Motor Activity / Q000187:drug effects; D018696:Neuroprotective Agents / Q000627:therapeutic use*; D051381:Rats; D017207:Rats, Sprague-Dawley; D027981:Symporters / Q000378:metabolism*; D000096922:K Cl- Cotransporters
D003975:Diazepam; D018696:Neuroprotective Agents; D027981:Symporters; D000096922:K Cl- Cotransporters
Diazepam; Hypoxic-ischemic; KCC2; Neuroprotective