RESUMEN
Hypermetioninemia is a group of rare diseases defined by plasma methionine elevation. The causes of hypermethioninemia include genetic and non-genetic factors. Affecting the transmethylation process in the metabolic pathway between methionine and homocysteine is the common inherited methylation disorders. Most patients had completely asymptomatic and only biochemical abnormalities,others with clinical symptoms were also non-specific, such as mental retardation, cognitive impairment, moderate hepatomegaly, dystonia, Marfans syndrome type,osteoporosis,and cardiovascular disease,and etc. A low methionine diet treatment is rec-ommended,the importance of neonatal screening is emphasized and the birth defects can be reduced through pre-natal diagnosis,but the necessity remains controversial. The long-term prognosis of this disorders is unknown,the plasma of methionine should be measured at regular intervals,and the individualized follow-up is very important.
RESUMEN
Objective To explore the inhibitory effects of endogenous hydrogen sulfide, a novel and important gas-eous transmitter generated in mammalian tissues mainly by cystathionine β-synthase (CBS) or cystathionineγ-lyase (CSE) on the apoptosis of the rat hepatic BRL cell line in physiological condition. Methods BRL cells were cultured, and divid-ed randomly into several groups in different phases of the experiment, including negative-siRNA (control) group, CBS siRNA (CBS 1~3) group and CSE siRNA (CSE 1~3) group, which were used to select the most efficient sequences of siRNAs at 48 or 24-hour transfection. Solution group and (CBS+CSE) siRNA group were added to detect the variation of apoptosis. The BRL cell line was observed and evaluated at 0, 4, 8, 12, and 24 hrs after siRNA transfection. When the mechanisms of the apoptosis were detected, CBS/CSE siRNAs were transfected individually or jointly into BRL cells, and compared with nega-tive-siRNA group to examine the variation. The genic and protein expression of CBS/CSE were detected by RT-PCR and Western blot assay. After transfection of CBS/CSE siRNA, the apoptosis of BRL cells was detected by Hoechst stain and flow cytometry (FCM). The mitochondrial membrane potential (MMP) changes were observed by fluorescent staining. Western blot assay was used to examine the protein expression of intracytoplasm cytochrome C (Cyt C) and cleaved-caspase 3. Re-sults CBS and CSE were observed in BRL cells. After transfection of CBS/CSE siRNA, endogenous H2S generation de-creased and the apoptosis of BRL cells increased. Accordingly, the expression of intracytoplasm-Cyt C and cleaved-caspase 3 increased. Conclusion The inhibition of endogenous H2S synthesis induced the apoptosis of BRL cells under physiologi-cal condition, which may be involved in mitochondrial pathway of apoptosis.
RESUMEN
Objective To establish hypoxic-ischemic brain damaged (HIBD) rat model,investigate whether H2S and cystathionine-β-synthase (CBS), the key enzyme for its generation, may be a mediator of electro-acupuncture(EA) stimulation treatment for HIBD. Methods Thirty-two healthy Sprague-Dawley neonatal rats were divided into four groups randomly: sham control group ( n = 8 ); sham + EA group ( n =8); HIBD control group ( n = 8); and HIBD + EA group ( n = 8 ). HIBD rat models were established on their 7-day-old. From the next day ,rats of sham + EA group and HIBD + EA group were electric stimulated 30 min daily for 14 d,BAIHUI and DAZHUI as the acupoints. Control ones were just fixed at the same time,without acupuncture. The rats were sacrificed on the 22 nd day, one day after the treatment course. Cortical H2S concentration was measured by sensitive sulphur electrode assay. The CBS protein expression was measured by western blot analysis and immunohistochemistry for localization. Results The concentration of cortical H2S in HIBD control group was (26. 83 ± 4. 31 ) nmol/mg protein, which was significantly higher than that of sham control group[(22. 78 ± 1.54) nmol/mg protein]( P ＜ 0. 01 ). The H2 S levels in HIBD + EA group and sham + EA group were ( 18.08 ± 2.71 ) nmol/mg protein and ( 18.91 ± 2. 78 ) nmol/mg protein, respectively. Compared with corresponding control group, they were much lower( P ＜ 0. 01 ). The expression of CBS protein in rats with EA stimulation decreased in cortex compared to corresponding control group( P ＜0. 05 ).Conclusion EA can down-resulate H2S/CBS pathway. This may be one of the mechanisms of how EA contributes to the recovery of brain damage.