| Objective:Insomnia is a sleep disorders in the occurrence and maintenance of the disorder as themain feature, mainly manifested as difficulty falling asleep, short sleep time, poor quality ofsleep, wake up early and daytime dysfunction, etc. Insomnia can be caused by many factors, itcan exist independently or appear together with other sleep disorders, department of internalmedicine disease or mental illness."Shu-Mian decoction" is professor Zhong-nan Wang’s patent medicine, clinical resultsare satisfactory. To further elucidate the mechanism of insomnia Curative effect and theprotective effect of cerebral neurons of "Shu-Mian decoction", this research study on theeffect of "Shu-Mian decoction" on content of5HT, DA in different brain regions, expressionchange of5HT1a,5HT2a, DRD2receptor mRNA, HPA axis hormones and anti-hippocampalneuronal apoptosis in insomnia model rats induced by PCPA experimentally.Methods:1.60Wistar rats were randomly divided into blank group, model group,"sleeps" highdose group, middle dose group, low dose group of "Shu-Mian decoction" and control groupwith18rats in each group, model by intraperitoneal injection of PCPA (400mg/kg in the8:00AM-09:00AM, for two consecutive days) consumption brain5HT and cause insomniamethod. Statistical analysis of sleep latency and sleep duration were did between the modelgroup rats and the normal control group rats in the first12hours after the secondintraperitoneal injection of PCPA with sodium pentobarbital sleep experiment (after the preexperimental sodium pentobarbital dose with30mg/kg) to evaluate model success; Observethe sleep latency and duration of sleep after treatment2. After modeling success of each group of mice given different treatments, the blankgroup and the model group were given physiological saline (1ml/100g)," Shu-Mian decoction" low dose group were given (6g/kg,1ml/100g),"Shu-Mian decoction " middle dose groupwere given (12g/kg, lml/100g)," Shu-Mian decoction " high dose group were give (24g/kg,1ml/100g), diazepam group given were given (0.0525mg/100g,1ml/100g) by gavagetreatment. After intragastric administration for7days for materials, the ELISA method wereused to test the5HT content in hypothalamus and hippocampus; qPCR (Real-time Quantitative PCR) technique were used for the measurement of5HT1a、5HT2a express in thehypothalamus and hippocampus.3. Molding and gastric perfusion method ibid, DA content in hypothalamus and striatumwere determined by ELISA method; DRD2receptor mRNA express in hypothalamus andstriatum were measured by qPCR technology.4. Determination of CRH content in hypothalamus and ACTH, CORT in serum ofinsomnia rats were tested by ELISA method5. Immunohistochemical method were used to observe the expression of BCL-2,BAXprotein in the hippocampus.Results:1. Compared with the blank control group, intraperitoneal injection of PCPA can makethe sleep latency prolonged and sleeping time shortened in rats(P<0.05);"Shu-Miandecoction" can reduce the trend of less weight increasing caused by insomnia; and the modelgroup sleeps" can relieve insomnia caused by reduced the trend of increasing weight loss(P<0.05); Compared with model group "Shu-Mian decoction" can shorten the insomnia ratssleep latency (P<0.05), increase the sleep time (P<0.05), played the effect on the treatment ofinsomnia.2. After treatment, compared with the model group，high dose group and middle dosegroup5HT content in hypothalamus and hippocampus of insomnia rats had significantlyincreased (P=0.011, P=0.049; P=0.005, P=0.037); compared with the model group，5HT1AmRNA express in hypothalamus and hippocampus was significantly increasing in the highdose group and middle dose group (P=0.007, P=0.046; P=0.005, P=0.027); compared with themodel group,5HT2A mRNA express in the hypothalamus and hippocampus statisticallydecreased in high dose group, and middle dose group rats (P=0.004, P=0.026; P=0.025,P=0.014).3. After treatment,compared with the model group, the content of DA in thehypothalamus was significantly decreased in the high dosage group and middle dosegroup(P=0.021, P=0.019); compared with the model group, the content of DA wassignificantly decreased in striatum in the high dosage group and middle dose group (P=0.046,P=0.007); compared with the model group, DRD2mRNA express in the hypothalamus was significantly decreased in the high dosage group and middle dose group (P=0.006, P=0.039);compared with the model group, DRD2mRNA express in the striatum was significantlydecreased in middle dose group.4. After treatment, compared with the model group, CRH in the hypothalamus wassignificantly decreased in the high dosage group and middle dose group (P=0.017, P=0.027);compared with the model group the contents of ACTH in the serum was significantlydecreased in the high dosage group and middle dose group (P=0.015, P=0.035); comparedwith the model group, content of CORT in serum was significantly decreased in middle dosegroup (P=0.016).5. After treatment, compared with the model group, the expression of BCL-2protein inrat hippocampus was significantly increased in high dosage group and middle dose group(P=0.015, P=0.035); compared with model group, the expression of BAX protein inhippocampus was significantly decreased in high dosage group and middle dose sleeps(P=0.011, P=0.042).Conclusion:1. Intraperitoneal injection of PCPA can prolonged the sleep latency and shortenedsleeping time in rats, so it is an effective method to make insomnia model;"Shu-Miandecoction " can improve the general condition of insomnia rats, reduce the trend of lessweight increasing caused by insomnia, can shorten the sleep latency and prolong the sleeptime, plays a therapeutic effect on insomnia.2. High dose group and middle dose group can significantly increase the5HT content inhypothalamus and hippocampus of insomnia rats, increasing5HT1A mRNA express inhypothalamus and hippocampus, decrease5HT2A mRNA express in the hypothalamus andhippocampus. It means the possible mechanism of therapeutic effect of "Shu-Mian decoction" may be caused by the increased express of5HT1A receptor and decrease express of5HT2Areceptor in the change of hypothalamus and hippocampus, resulted in the increase of5HTcontent of it.3. The high dosage group and middle dose group significantly decreased DA in thehypothalamus and striatum; High dosage group and middle dose group significantly decreasedthe DRD2mRNA express in the hypothalamus and striatum. It means the possible mechanism of therapeutic effect of "Shu-Mian decoction " may be caused by decreased the DRD2mRNAexpress in the hypothalamus and striatum and result in the decreased DA of hypothalamus andstriatum4. The high dosage group and middle dose group significantly decreased CRH in thehypothalamus and ACTH、CORT in the serum. It means the possible mechanism oftherapeutic effect of "Shu-Mian decoction " may be caused by decrease of CRH, ACTHCORT, alleviate the excessive state of HPA axis caused by. Insomnia.5. High dosage group and middle dose group can significantly increase the expression ofBCL-2protein in rat hippocampus and decreased the expression of BAX protein inhippocampus, It means "Shu-Mian decoction " can change the cell apoptosis factor expressionto protect hippocampal neurons. It May be due to the therapeutic effect of "Shu-Miandecoction " on insomnia. |
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