Effects of Kaixin Jieyu Decoction () on behavior, monoamine neurotransmitter levels, and serotonin receptor subtype expression in the brain of a rat depression model / 中国结合医学杂志

<p><b>OBJECTIVE</b>To determine the mechanisms underlying the anti-depressant effects of Kaixin Jieyu Decoction (, KJD) by investigating the effects of KJD on behavior, monoamine neurotransmitter levels, and serotonin (5-HT) receptor subtype expression in the brain in a rat model of depression.</p><p><b>METHODS</b>The rat depression model was established using chronic unpredictable mild stress (CUMS). Forty-eight Sprague Dawley rats were randomly divided into control, depression model (CUMS), CUMS+KJD (7.7 g/kg(-1)·d(-1) of crude drug), and CUMS+fluoxetine (2.4 mg/kg(-1)·d(-1)) groups (n=12 in each group), and the treatments lasted for 21 days. We regularly evaluated body weight, sucrose consumption, and horizontal and vertical activity scores in open-field tests. The content of the monoamine neurotransmitters 5-HT, norepinephrine (NE), and dopamine (DA) and the DA metabolite homovanillic acid in the cerebral cortex, and 5-HT1A and 5-HT2A receptor mRNA in the cerebral cortex and the hippocampus, were determined respectively by high-performance liquid chromatography-coularray electrochemical detector and real-time polymerase chain reaction.</p><p><b>RESULTS</b>Compared with the control group, CUMS rats showed a variety of depression-like behavioral changes, including a significant reduction in body weight, sucrose consumption, and horizontal and vertical activity scores in open-field tests (P<0.05 or P<0.01), and a significant decrease in 5-HT and NE levels and 5-HT2A receptor mRNA expression. In contrast, they showed a significant increase in 5-HT1A receptor mRNA expression in the cerebral cortex. In the hippocampus, 5-HT1A receptor mRNA expression was lower whereas 5-HT2A receptor mRNA expression was higher than in the control group (P<0.05 or P<0.01). Treatment with KJD or fluoxetine partially attenuated these changes (P<0.05 or P<0.01).</p><p><b>CONCLUSION</b>KJD could normalize the levels of 5-HT and NE and adjust the balance of 5-HT1A and 5-HT2A receptor expression in rat cerebrum, and this may be one of mechanisms of antidepressant effects of KJD.</p>

Effects of environmental enrichment on maternal behavior of rats with early life stress / 中华行为医学与脑科学杂志

Objective To investigate the improve effects of environmental enrichment on maternal behavior in the adult female rats thatexperienced early-life stress,and to explore the molecular biological mechanism.Methods The newborn Wistar rats were divided into two groups randomly,the early life stress group and the control group.Pups from the former were separated from their mothers once per day for 4 hours from PND 2-21 (day of birth was considered PND =0),and suffered vicious stimulus during the daily 4 hours maternal separation from PND 8-21.On PND 22(weaning),pups from the early life stress group were divided into two groups:12 female pups were reared singly,refered to as isolated-rearing group; another 12 were placed in a big cage that was filled with all sorts of rodents toys,refered to as enriched-rearing group; control female pups were placed in groups of 4 per cage.Mating with male rats when female pups reached PND66,then being placed in cages singly when they were detected pregnant.Maternal behavior was evaluated on the second and sixth day after delivery,and the hypothalamic OTR mRNA levels were examined by RT-PCR.Results The results of maternal behavior observation showed that the total time of retrieve and the latency of licking were significantly longer in isolated-rearing rats compared with the normal and the enriched-rearing rats (P ＜ 0.01) ; total time of licking in enriched-rearing rats was longer than the normal and the isolated-rearing rats(P＜0.01) on the sixth day after delivery;and the score of the nest building was obviously lower in isolated-rearing rats,especially on the sixth day after delivery(P ＜ 0.01).Hypothalamic OTR mRNA levels in the isolated-rearing rats showed strongly decreased compared with the normal rats(P＜0.01),but which close to normal levels in enriched-rearing rats.Conclusion Early life stress can damage some components of maternal behavior,such as retrieval,licking,nest building,but it has no obvious effect on crouching behavior; it also decreases the hypothalamic OTR mRNA levels of the adult female rats; as a kind of early environmental intervention,groups-rearing could make hypothalamic OTR mRNA levels of the female rats that subjected to early life stress restore to normal,and thus reverse the disruptions of stress in maternal behavior.

ZMS regulation of M_2 muscarinic receptor stability mediated by de novo synthesis of protein / 上海交通大学学报（医学版）

Objective To explore the mechanism of ZMS regulation of M_2 muscarinic receptor mRNA expression. Methods In vitro cultured CHOm2 cells were divided into ZMS 1 group (treatment with 1 × 10~(-5) mol/L ZMS for 24 h), ZMS 2 group (treatment with 1 × 10~(-5) mol/L ZMS for 24 h and 1 μg/mL cycloheximide for 12 h) and ZMS 3 group (treatment with 1 μg/mL cycloheximide for 4 h and 1 × 10 ~(-5) mol/L ZMS for 24 h), and their corresponding control groups were also established (substitution of ZMS by DMSO). Actinomycin D was added to cultured CHOm2 cells of each group to inhibit the synthesis of mRNA. CHOm2 cell samples were taken at different time points, the relative expression of M_2 receptor mRNA was detected by Real-time PCR, and half life of M_2 receptor mRNA was calculated. Results Compared with corresponding control groups, the half life of M_2 receptor mRNA of CHOm2 cells in ZMS 1 group and ZMS 2 group was significantly prolonged [(4.75h± 0.54) h vs (2.13 ±0.23) h, P<0.05; (5.43 ±1.13) h vs (2.46 ±0.09) h, P<0.05]. There was no significant difference in half life of M_2 receptor mRNA of CHOm2 cells between ZMS 3 group and its corresponding control [ ( 3.06 ±0.23) h vs (3.00 ± 0.20) h, P > 0.05]. Conclusion De novo protein synthesis is required for the enhancement of M_2 receptor mRNA stability regulated by ZMS.

Effects of Electroacupuncture on Expression of ?-opioid Receptor mRNA of Morphine Withdrawal Rat / 中国中医药信息杂志

Objective To research the mechanism of electroacupuncture(EA) improving morphine abstinence syndrome in rats.Methods The model of the rat with physical morphine abstinence syndrome was established.The expression of ?-opioid receptor mRNA in the brain tissue of morphine withdrawal rats was examined by RT-PCR,and effects of EA at Zusanli(ST36) on the body weight and expression of ?-opioid receptor mRNA of morphine withdrawal rat were observed.Results The body weight of the EA group was increased compared with the morphine abstinence group(P

Effects of Isoflavones Supplemented Diet on Lipid Concentrations and Hepatic LDL Receptor mRNA Level in Growing Female Rats

The purpose of this study was to examine the impact of isoflavones on lipid concentrations and hepatic LDL receptor mRNA level in growing female rats. Twenty four rats (body weight 75 +/- 5 g) were randomly assigned to one of two groups, consuming control diet or isoflavones supplemented diet (57 mg isoflavones/100 g diet). All rats has been fed on experimental diet and deionized water ad libitum for 9 weeks. The concentration of triglyceride and total cholesterol were measured in serum and liver. Serum HDL cholesterol was measured. Hepatic LDL receptor mRNA level was tested by RT-PCR. Supplementation of isoflavones did not affect weight gain, mean food intake and food efficiency ratio. Serum total cholesterol and non-HDL cholesterol of isoflavones supplemented rats were significantly lower than those of control rats (p < 0.05). But hepatic cholseterol level was not influenced by supplementation of isoflavones. Hepatic LDL receptor mRNA level not significantly different between control group and isoflavones supplemented group. Therefore, isoflavones may be beneficial on serum cholesterol and non-HDL cholesterol lowering in growing female rats.

Changes of beta-Adrenergic Receptor mRNA in the Visual Cortex and Superior Colliculus of Monocular Deprivated Rat

PURPOSE: To investigate the change of the Beta adrenergic system in the rat visual cortex and superior colliculus after visual deprivation during a critical period of postnatal development. METHODS: The changes of beta 1 and beta 2 adrenergic receptor mRNA were investigated by using northern blot analysis in the rat visual cortex and superior colliculus. The right eyelid of visually deprived rat was sutured at the 10th postnatal days. After visual deprivation for 4 weeks, the rat were sacrificed and the visual cortex and superior colliculus tissues were removed for analysis. RESULTS: Beta 1 and beta 2 adrenergic receptor mRNA expression was decreased in the contralateral visual cortex to the deprived eye. In the superior colliculus, beta 2 adrenergic receptor mRNA expression increased in both sides, but a much greater increase was shown in the ipsilateral superior colliculus than the contralateral side. CONCLUSIONS: These results suggests that visual deprivation during a critical period of postnatal development influences the beta adrenergic system in the rat visual cortex and superior colliculus.

Changes of mRNA Expression of Dopamine Receptor in the Visually Deprivated Rat Striatum

PURPOSE: To investigate the changes of the dopaminergic system in rat striatum after visual deprivation during a critical period of postnatal development. METHODS: The changes of dopamine D1 and D2 receptor (Rc) mRNA were investigated by using Northern blot analysis, in situ hybridization in the rat striatum. The right eyelid of visually deprivated rat was sutured at 10 postnatal days. After visual deprivation for 4 weeks, rats were sacrificed and striatum tissues were removed for analysis. RESULTS: By Northern blot analysis and in situ hybridization, decreased expression of D1 and D2 Rc mRNA was noted in the ipsilateral striatum to the deprivated eye. CONCLUSIONS: These results have shown that visual deprivation during a critical period of postnatal development influences the dopaminergic system in the striatum.

Regulation of gonadotropin releasing hormone receptor mRNA expression in cultured rat granulosa cells

The homologous regulation of pituitary Gonadotropin Releasing Hormone Receptor (GnRH-R) mRNA expression by GnRH has been well demonstrated. However, the regulation of the ovarian GnRH-R is poorly understood. The present study was performed to demonstrate the presence of GnRH transcripts in addition to GnRH-R mRNA and the regulation of GnRH-R mRNA expression in the granulosa cells isolated from small antral follicles. The GnRH and GnRH-R mRNA levels were determined by a competitive reverse transcription-polymerase chain reaction (RT-PCR). The granulosa cells were obtained from immature rats implanted with diethylstilbestrol for 3 days. When GnRH transcript expression was examined in isolated granulosa cells by RT-PCR, the PCR products showed two bands. The larger band contained intronic sequences and the smaller band was a fully processed GnRH gene transcript identical to hypothalamic GnRH. This suggests that authentic GnRH gene transcripts are expressed in ovarian granulosa cells and may act on the granulosa cells in a paracrine or autocrine manner. Since GnRH action in the granulosa cells is mediated by specific GnRH-R, it is of interest to examine whether GnRH-R is synthesized in the granulosa cells. When the granulosa cells were cultured in media only, GnRH-R mRNA levels increased abruptly within 3 h and gradually decreased thereafter during the 24 h culture period. However, GnRH itself did not alter the GnRH-R mRNA expression levels in cultured granulosa cells. Interestingly, treatment with FSH decreased the GnRH-R mRNA levels in a dose-dependent manner. A time-course analysis revealed that the GnRH-R mRNA levels were significantly lower up to 9 h after FSH treatment, and returned to the basal level between 12 h-24 h. Activation of adenylate cyclase with forskolin also decreased the GnRH-R mRNA levels. It is therefore concluded that in the granulosa cells of the small antral follicles GnRH-R mRNA expression was not homologously regulated by GnRH, while FSH may negatively regulate GnRH-R mRNA expression in the granulosa cells possibly through a cAMP-protein kinase A pathway.

Detection of the Expression of GnRH and GnRH Receptor mRNAs and Effect of GnRH on Prostate Cancer Cell Proliferation / 대한비뇨기과학회지

Prostatic cancer is the typical hormone dependant cancer and several kinds of hormones are used for the treatment of prostate cancer. Since Harris had proposed the hypothalamo-pituitary gonadal axis, the hypothalamus is believed the exclusive organ producing GnRH. According to the recent researches, several organs were proved to be the extrahypothalamic GnRH sources in human and animal. In this research, the expression of the GnRH and GnRH receptor mRNA, detection of the GnRH which prostate cancer cell produced and effect of the GnRH on the prostate cancer cell proliferation using three human prostate cancer cell lines, ALVA 41, ALVA 101 and DU-145 were studied. In Situ Hybridization method was used for the detection of the expression of the GnRH and GnRH receptor mRNA. The charcoal stripped serum and high performance liquid chromatography were used for the detection of the GnRH produced from prostate cancer cells. Thymidine incorporation assay was used for the evaluation of the effect of the GnRH on the prostate cancer cell proliferation. The GnRH mRNA were detected in 96.7% of ALVA 41, 91.5% of ALVA 101 and 95.3% of DU-145 and GnRH receptor mRNA expression signals were detected in almost all of the examined prostate cancer cells, more than 95%, in three cell lines. The number of signals of the GnRH receptor mRNA were more than GnRH mRNA. The GnRH produced from the rostate cancer cell was detected at culture medium with retention time 19.40 minutes. The cancer cells cultured with peptide hormone deficient medium using charcoal stripped serum showed more than 20% growth retardation to the cancer cells grown at the medium used normal serum. The treatment of the GnRH on the cancer cells growing at the peptide hormone deficient medium showed statistically insignificant dose dependant growth retardation. The RESULTS of our research showed that the human prostate cancer cells, including two hormone refractory prostate cancer cell lines, produce the GnRH and the GnRH receptor in the same cell which could be suggest that the role of GnRH produced from the prostate cancer cell would be autocrine action. And the prostate cancer cell growth was down regulated by unknown complex of various peptide hormones and the GnRH does not has the significant effect on the proliferations of the prostate cancer cells. With those RESULTS we obtained in this research and other's data, it seems that there is a system that contains production of GnRH and GnRH receptor and metabolic mechinary within prostate cancer cell. And there should be the some changes in the hypothalamo-pituitary gonadal axis and the mechanism using GnRH analogues for the treatment of prostate cancer aside from central mechanism.

Expression of Gonadotropin-Releasing Hormone Receptor in Human Uterine Endometrial and Ovarian Tissues / 대한암학회지

PURPOSE: Gonadotropin-releasing hormone (Gn-RH) can cause regression of hormonedependent human tumors, including uterine endometrial and ovarian carcinomas. These effects were thought to be mediated through the inhibition of gonadotropic and steroid hormone from the hypothalamus. But, in addition to its classic hypophysiotropic action, Gn-RH might play a role as a modulator of activity in the brain and many peripheral organs. It has been reported that this analog has a direct inhibitory effect on the tumor and that the specific binding sites for Gn-RH were demonstrated in certain tumors responsive to Gn-RH. In support of a possible clinical use of Gn-RH analogs in the treatment of the endometrial and ovarian carcinomas, we tried to find out whether Gn-RH receptors are present on hormone dependent tumors. MATERIALS AND METHODS: We have studied endometrial and ovarian tumor specimens and established uterine endometrial and ovarian carcinoma cell lines for the presence of Gn-RH receptor by the detection of its messenger ribonucleic acid (mRNA). We also compared the results obtained from tumor tissue specimens with the results from their corresponding normal tissues. Gn-RH receptor mRNA was determined by reverse transcription-polymerase chain reaction using oligonucleotide primers synthesized according to the published human Gn-RH receptor sequence. RESULTS: Gn-RH receptor mRNA was detected in all normal endometrium and abnormally proliferative endometrium presenting dysfunctional bleeding, but not all in endometrial carcinomas (83%). Tumor stage and histologic grading had no relationship with receptor positivity. And, Gn-RH receptor mRNA was detected in less than 40% in normal myometrium and myomas. Gn-RH receptor expression was detected in same frequencies (86%) in normal ovarian tissues and ovarian carcinomas. Receptors were detected in a high proportion of the specimens from epithelial carcinomas (92%) and stromal tumors (100%) of the ovary. But, Gn-RH receptor was not detected in germ-cell derived tumors of the ovary. Established endometrial carcinoma (CUME-1) and epithelial ovarian carcinoma (CUMO-2) cell lines also demonstrated Gn-RH receptor mRNA, respectively. CONCLUSIONS: The expression of Gn-RH receptor raises the possibility that Gn-RH may play a direct regulatory role in the growth of hormone-dependent normal tissues and their respective tumors, and provides a possible point of attack for therapeutic approaches using Gn-RH analogs in endometrial and ovarian malignancies.

Expression and Distribution of Beta 2 Adrenergic Receptor mRNA in the Rat eye

Beta adrenergic receptors(beta-ARs) were detected in retina, trabecular meshwork, iris and ciliary body in eye of human, cat, rabbit, and bovine using in vitro autoradiography and majority of the beta-ARs found in eye are the beta2 subtype. Recently, the beta2-AR gene has been cloned from hamster, human, rat and monkey using molecular biological methods. Expression of beta2-AR mRNA were demonstrated in smooth muscle, kidney, ovary, brain, adipose tissue, heart, epithelial cells, thymus, lung and liver. However, studies about expression and distribution of beta2-AR mRNA in the eye have not been done yet. Author have characterized the expression of beta2-AR mRNA in rat eye using in situ hybridization with 35S-UTP riboprobe. beta2-AR mRNA was expressed in corneal epithelium and stroma, ciliary epithelium, vessels of ciliary body, choroidal vessel, and retina. In contrast it was not expressed in iris and sclera in the rat eyes. These results support the hypothesis that beta2-AR mRNA may be synthesized in the various ocular tissue and its characterized distribution suggests partially that beta2-ARs are related with aqueous production and blood supply of the eye.

Expression of Epidermal Growth Factor Receptor mRNA by In Situ Hybridization in Normal and Abnormal Thyroid Tissue / 대한내분비학회지

Growth factors are polypeptide molecules that regulate cell growth and function by binding with high affinity to specific receptor molecules in the plasma membrane and stimulating receptor mediated action of intracellular signal transduction pathway.Epidermal growth factor(EGF) and their receptors(EGFR) regulate normal cellular growth, proliferation, and differentiation of various cells in vivo and in tissue cultures. And also may contribute directly to oncogenesis.Overexpression of EGFR and autocrine stimulation of growth involving this receptor system has been identified in several types of human neoplasia. There is evidence that the EGF and receptor system is involved in the regulation of follicular cell growth in the thyroid gland especially with immunohistochemical technic. But there was a challenge about the validity of previously performed immunohistochemical studies.In the study we investigated the relationship between EGFR mRNA expression and tumorigenesis by rapid in situ hybridization method. Formalin-fixed, paraffin embedded tissue sections of 10 normal, 17 nodular hyperplasia, 6 follicular adenoma, and 15 papillary cancer were examined. The results were as follows:1) EGFR mRNA positivity were 20%(2/10) in normal thyroid, 70%(12/17) in nodular hyperplasia, and 100% in follicular adenoma and papillary cancer.2) There was a significantly increased EGFR mRNA expression in papillary cancer compare to normal and nodular hyperplasia(p<0.05). But no difference was found with papillary cancer and follicular adenoma.3) There was a significantly increased EGFR mRNA expression in follicular adenoma compare to normal (p<0.05). But no difference was found with follicular adenoma and nodular hyperplasia. These results suggest that an overexpression of EGFR mRNA may play an important role in the tumorigenesis of thyroid tissue.

EXPRESSION OF T_3 RECEPTOR c-erbA? mRNA IN EXPERIMENTAL HYPERTHYROID AND HYPOTHYROID RATS / 中华内分泌代谢杂志

The rat c-erbA? cDNA fragment was used as probe to investigate the expression of T3 receptor gene in brain, heart and kidney in hyperthyroid and hypothyroid rats. There were three types of c-erbA? mRNA-6. 0kb, -5. 0kb and -2. 0kb in all examined tissues. In heart and kidney, the levels of c-erbA? mRNA decreased in hyperthyroid rats and increased in hypothyroid rats. It might be a compensatory mechanism. In brain, however, the level of c-erbA? mRNA was unaffected by thyroid status. The results suggest that the T3 receptor gene may have an auto-regulatory mechanism in rat heart and kidney under different thyroid status.