Qing Kai Ling Injection On The Regulatory Mechanism Of Ion Homeostasis In Ischemic Brain Damage

Ischemic cerebrovascular diseases are common and frequently occurring disease, and have high case fatality rate and mutilation rate. They have threatens human healty seriously and In the last few years,all kinds of ions has been an important topic in medical domain of ischemic cerebrovascular diseases.In order to provide more modern biological basis of unbalanced nutrient qi and defensive qi caused by collaterals stasis resistance,our experiment is guided by the pathogens is theory of 'toxin hurts brain collaterals' and by the cut in point of ions, to study the damage of ion homeostasis and the effect of Qingkailing injection(QKL)for investigating the mechanisms on how Chinese herbs keep the ion homeostasis caused by cerebral ischemia and protecting neurons by detoxicating, dredging collaterals and harmonizing nutrient qi and defensive.. The purpose of our research is to supply meaningful ideas for the approach and targets of Chinese herbs therapy on diseases of the central nervous system.Objective1. To ascertain the damage of ion homeostasis caused by cerebral ischemia in rat's brain and the protection of QKL to neurons in rats. To further verify the variation characteristics of zinc ion in ischemic injury and QKL on its influence.2. To simply clarify the mechanism of the damage of ion homeostasis in ischemic injury caused by changed expression of methallothionein(MT) which maybe regulated by the status of astrocyte cells.and overloading Calcium and clarify pharmacological mechanism. of QKL injection.Method1. To make focal cerebral ischemia injury model by MCAO method, assess and filter the MCAO rats by neurological scores and to observe morphological change by light microscope and electronic microscope.2. To detect the concentration of K+, Na+, Cl-, Ca2+,Zn2+, Cu2+, Mg2+, Fe2+ in brain serum and CSF of rats in N group, M group and T group by automatic biochemistry analyzer and Inductively coupled plasma mass spectrometer (ICP-MS). To detect the level of K+, Na+, Ca2+,Zn2+,Cu2+,Mg2+,Fe2+ in QKL injection by ICP-MS. To observe morphological change of mitochondria in neurons,astrocytes by electronic microscope.3. To observe the expression of Zn2+ in brain tissue of rats in N group, M group and T group throug ivZnSAMG method by light microscope.4. To observe the expression of glial fibrillay acidic protein(GFAP in brain tissue of rats in N group, M group and T group by immunohistochemistry method to show the status of the originate cells—astrocyte (Ast). To observe the expression of methallothionein(MT) in brain tissue of rats in N group, M group,T group and MK group by immunohistochemistry method. To detect the concentration of Ca2+, Zn2+in brain of rats in N group, M group,T group and MK group by Inductively coupled plasma atomic emission spectrometer (ICP-AES)Result1.24h after MCAO,a lot of cortex nurons were lost, normal structure were disordered, narrowing nurons and swelling astocyte occurred in cortex area. but nurons and asrocytes were restored after rats were given QKL injection. The results show consistency with neurobehavioral scores.2. From high to low,the content of each element in the QKL is Na+, K+, Ca2+, Zn2+ Fe2+, Mg2+, Cu2+.24h after MCAO, it can be seen swelling mitochondria of neurons and Ast in rat's cortex area,but mitochondria became stick or domed after rats were given QKL injection.QKL could restored many ions'homeostasis,for example, K+, Ca2+,Zn2+ in the brain; K+, Na+, Cl-, Ca2+, Mg2+, Fe2+ in the serum; K+ Na+ in the cerebrospinal fluid.3.24h after MCAO, the express strength of Zn2+ in rats'brain significantly drops (P<0.05), while it raises up after rats were given QKL injection.4.24h after MCAO, the change of Zn2+ in rats'brain is contrary to Ca2+. They have negative correlation. After rats were given calcium channel blockers, the change of Zn2+ in rats'brain is still contrary to Ca2+. The content of MT in brain of model group significantly raises up than normal group(P<0.01), while it of QKL group significantly drops (P<0.01) and it of MK group significantly drops(P<0.01),24h after MCAO, the content of GFAP in rats' brain significantly raises up(P<0.01), while it significantly drops (P<0.01) after rats were given calcium channel blockers.Conclusion:1.24h after MCAO, the neurons of rats obviously damage, QKL could protect brain cells from injury.2.24h after MCAO, rats' brain, serum, cerebrospinal fluid have metabolic dishomeostasis, QKL could affect metabolic dishomeostasis during ischemia cascade process by providing energy and various ions for brain cells.3. Cerebral ischemic injury reduced brian's Zn2+ content,QKL could increase Zn2+ content.4. After cerebral ischemic injury, the content of Zn2+ significantly drops due to excessive expression of MT. Calcium overload and increasing astrocytes could promote MT expression.QKLcould maintain Zn2+.homeostasis. by easing calcium overload, adjusting astrocytes expression.