1.Effect of Gypsophila elegans isoorientin on proliferation and apoptosis of human HepG2 cells
Jinlan NIE ; Quanfang HUANG ; Shimei TAN ; Xing LIN
Chinese Journal of Pharmacology and Toxicology 2016;30(9):928-934
OBJECTIVE To investigate the effect and underlying mechanism of Gypsophila elegans isoorientin on the proliferation and apoptosis of human HepG2 cells. METHODS HepG2 cells were treated with isoorientin 5,10,20,40,80 and 160μmol?L-1 for 24,48 and 72 h,respectively. Cell survival was analyzed by MTT assay. HepG2 cells were treated with isoorientin 5,10 and 20μmol?L-1 for 48 h before the lactate dehydrogenase(LDH)level was detected. After treatment with isoorientin for 24 h, the variation of reactive oxygen species (ROS) was monitored by a fluorescence probe H2DCF-DA. HepG2 apoptosis and mitochondria membrane potential(MMP)were evaluated by flow cytometry. The activi?ties of caspase 3,8 and 9 were determined by colorimetry. The mRNA expression of Bcl-2 and Bax was determined by RT-PCR,and the protein expression of Bcl-2, Bax and cytochrome c was detected by Western blotting. RESULTS Isoorientin(5-160μmol?L-1)inhibited HepG2 cell survial in a concen?tration-dependent manner,the 50%inhibitory concentration(IC50)was 62.7±9.1,47.2±11.4 and(18.2± 7.5)μmol?L-1 after treatment with isoorientin for 24,48 and 72 h,respectively. Compared with the cell control group,treatment with isoorientin 5,10 and 20μmol?L-1 significantly increased the LDH level and cell apoptosis rate(P<0.05). Moreover,isoorientin 10 and 20μmol?L-1 notably increased the production of ROS,decreased the MMP(P<0.05),and increased the activities of caspase 3 and 9. RT-PCR analysis and Western blotting showed that isoorientin significantly decreased the mRNA and protein expressions of Bcl-2,decreased the mRNA and protein expressions of Bax(P<0.05),and inhibited the protein expression of cytochrome c(P<0.05). CONCLUSION Isoorientin inhibits HepG2 cell prolif?eration,but promotes cell apoptosis,which is closely related to the regulation of the mitochondrial apoptosis pathway.
3.Weight-reduction Effect of Shanzha Xiaozhi Capsules on Obesity Patients with Phlegm-Dampness Constitution
Qijun LIANG ; Chenming HU ; Jinlan LAI ; Rong HUANG ; Ruizhu CHEN ; Huiping LI ; Dongcai LI ; Shouyi YU
Journal of Guangzhou University of Traditional Chinese Medicine 2016;33(5):625-629
Objective To investigate the effect of Shanzha Xiaozhi Capsules (SXC) on body weight, metabolic indexes, body fat accumulation and distribution of obesity patients with phlegm-dampness constitution. Methods Fifty obesity patients with phlegm-dampness constitution were randomized into control group and medicine group, 25 cases in each group. The control group was given lifestyle instructions for diet and exercises, and the medicine group was treated with SXC orally besides the instructions for lifestyle. The treatment for both groups covered 12 weeks. Before and after treatment, we observed the outcomes including scores of phlegm-dampness, body weight, waist circumference, hip circumference, blood pressure, fasting blood glucose, serum lipid profile, urine acid, body fat accumulation and distribution, and visceral fat accumulation. Results(1) The scores of phlegm-dampness were decreased significantly in both groups after treatment (P<0.05 or P<0.01), and the decrease was obvious in the medicine group(P<0.01).(2) Body weight and BMI were decreased significantly in both groups after treatment(P<0.01), and the decrease was obvious in the medicine group(P<0.05). Waist circumference was decreased significantly in the medicine group(P <0.05) but stayed unchanged in the control group(P>0.05). Hip circumference was decreased significantly in both groups(P <0.05 or P<0.01) , but the difference between the two groups was insignificant(P>0.05).(3) Systolic pressure and diastolic pressure were decreased significantly in both groups (P<0.01). But the difference of blood pressure decrease between the two groups was insignificant(P > 0.05).(4) Fasting blood glucose, 2-hour blood glucose after oral glucose tolerance test, serum lipid profile and urine acid remained unchanged in both groups after treatment (P>0.05). (5) The general body fat accumulation of both groups was decreased significantly after treatment (P <0.05 or P<0.01), and the decrease was obvious in the medicine group(P<0.01). Body fat accumulation in legs, trunk and gynoid region were decreased significantly in both groups after treatment(P<0.05 or P<0.01), but the difference of fat accumulation decrease between the two groups was insignificant(P>0.05). Waist fat accumulation had no significant decrease in the control group (P > 0 . 05), but was decreased in the medicine group (P<0.01). Visceral fat accumulation was decreased significantly in both groups after treatment(P<0.01), and the decrease was obvious in the medicine group (P<0.01). Conclusion In addition to its contribution to body weight loss, SXC also contribute to the reduction of visceral fat accumulation in obesity patients with phlegm-dampness constitution .
4.Effects of Flow Components C6 and C7 in N-butyl Alcohol Extract from the Leaves of Cestrum nocturnum on the Proliferation and Apoptosis of Human Gastric Cancer Cell SGC7901
Dengpan WU ; Wen LIAO ; Jinlan HUANG ; Wenya CHEN ; Yulu HE ; Shengrui ZHANG ; Zhenguo ZHONG
China Pharmacy 2015;26(31):4342-4344
OBJECTIVE:To study the effects of flow components C6 and C7 in n-butyl alcohol extract from the leaves of Ces-trum Nocturnum(CN)on the proliferation and apoptosis of human gastric cancer cell SGC7901. METHODS:C6 and C7 were ob-tained by using different ratio of chloroform and methanol(1:9,1:7)to the gradient elution of CN leaves. After cultured with 0 (blank control),5,10,20,40,80 μg/ml C6 and C7 for 72 h,inhibitory effect of C6 and C7 on the proliferation of SGC7901 was determined by MTT assay. Inhibitory rate and IC50 were calculated. After SGC7901 were cultured with 10 μg/ml C6 and C7 for 72 h,colony formation assay was utilized to detect the effects of C6 and C7 on the cell colony formation,and the rate of colony for-mation was calculated. In addition,Wright/Giemsa and Hoechst33258/PI staining assay were used to observe the change of cytomor-phology. RESULTS:MTT showed that C6 and C7 had inhibitory effect on the proliferation of SGC7901 to different extent;inhibi-tory rates were 22.1%-80.0% and 19.6%-79.7%,and IC50 were 16.4,18.05 μg/ml,respectively. Compared with blank control group,colony formation rate of C6 and C7 group decreased,with statistical significance(P<0.05). The number of apoptotic cells was more in treatment group than in other groups. CONCLUSIONS:Flow components C6 and C7 in n-butyl alcohol extract from the leaves of CN can inhibit the proliferation of SGC7901 cells and induce the apoptosis of them.
5.Clinical and genetic analysis of a family with familiar acute necrotizing encephalopathy due to mutation in the RANBP2 gene
Jinlan ZHU ; Tieshuan HUANG ; Jing DUAN ; Dong CUI ; Jialun WEN ; Jianxiang LIAO
Chinese Journal of Applied Clinical Pediatrics 2015;(21):1672-1675
Objective To analyze the clinical manifestations of familial acute necrotizing encephalopathy (ANE)and to improve the recognition of this disease. Methods The clinical data of a 25 - month - old girl with fa-milial and recurrent ANE with evidence of mutation in the RANBP2 gene were collected and analyzed,and the gene examination of their family members was performed. Results A previously healthy girl experienced recurrent ANE epi-sodes at the ages of 8 months,18 months and 25 months,respectively. At each beginning of each episodes the patient presented with lethargy and tremor of limbs following febrile illness of 3 - 4 days,even developed coma and convulsions in the last time. Brain magnetic resonance imaging showed bilateral and high T2 signal changes in thalamus,cerebellum and hippocampus. Abnormal signals also appeared in the brainstem,claustrum,corpus scallosum and cortex(temporal, parietal and cingulate)also appeared abnormal signals. Spinal MRI showed spinal cord involvement. The girl recovered after her first episode;she could speak but could not walk steadily after the second time;after the third episode,al-though she regained consciousness from coma,she could no longer speak or walk. The patient's sister died of encephali-tis at the age of 18 months. Her paternal uncle had suffered from dysnoesia from meningitis at his 17 months of age. The patient and her grandmother,father,uncle and one of her aunts harbored a mutation(c. 1754C ﹥ T)in RANBP2 gene. Conclusions Familial ANE has typical clinical manifestations and characteristic MRI findings. The patient with recur-rent history,especially with positive family history,should have the mutation in RANBP2 gene detected earlier in order to clarify the diagnosis of ANE.
6.Correlation Analysis between Serum Complement C3,C4 Levels and HDL-C in Patients with Coronary Atherosclerotic Heart Disease
Ruiting LIN ; Bo ZHANG ; Canxia HUANG ; Runlu SUN ; Weifeng LU ; Jinlan BAO ; Yuling ZHANG
Journal of Sun Yat-sen University(Medical Sciences) 2017;38(1):72-77
Objective]The aim of this study was to investigate the level of serum C3,C4 and HDL-C in patients with coronary heart disease and the correlation between C3,C4 and HDL-C in patients with coronary heart disease.[Methods]We collected 251 cases of patient diagnosed with coronary artery disease by coronary angiography in Sun Yat-sen Memorial Hospital ,Sun Yat-Sen University from 2015-12 to 2016-07 and collected over our Boji Medical Center healthy people in 214 cases. These patients were divided into acute coronary syndrome group with 180 cases and stable coronary heart disease group with 71 cases. Each test results was adopted from clinical laboratory of Sun Yat-Sen Memorial Hospital ,Sun Yat-Sen University.[Results]Compared with the healthy control ,the difference of serum C3,C4 and HDL-C from acute coronary syndrome group and stable coronary heart disease group,was statistically significant(P<0.05). However,in patients with acute coronary syndrome and stable coronary artery disease groups,there was no significant correlation(P > 0.05)between C3,C4 and HDL-C. In healthy group,complement C3 negatively correlated with HDL-C,the difference was statistically significant(P<0.05).[Conclusions]In patients with coronary heart disease, the level of C3 and C4 increased,while the level of HDL-C decreased ,and inflammation may affect the relevance judgments between complement and HDL-C.
7.Application of three-dimensional arterial spin labeling for cerebral blood flow in migraine without aura
Yongqiang YE ; Jun ZHANG ; Jie TANG ; Yu SHEN ; Jinlan HUANG ; Jing CHEN ; Xiaohui WANG
Chinese Journal of Neurology 2017;50(1):40-43
Objective To investigate the cerebral blood flow in migraine without aura with three-dimensional arterial spin labeling technology .Methods From January 2013 to February 2016, magnetic resonance perfusion imaging was operated in 20 migraine patients without aura and 20 healthy controls by using arterial spin labeling, to determine their regional cerebral blood flow (rCBF) during migraine attacks and attack-free period.The significant difference was analyzed in rCBF between the two groups .Results The rCBF was significantly lower at the headache side during migraine period ( ( 52.77 ±5.97 ) ml? 100 g-1? min-1 ) than symmetrical side in the migraine patients without aura ((62.72 ±6.65) ml? 100 g-1?min-1 , t=3.780,P<0.01).The rCBF of temporosphenoid lobe in headache side in the migraine patients without aura ((53.97 ±5.87) ml? 100 g-1? min-1) was significantly lower than corresponding region in the control group ((59.21 ±2.35) ml? 100 g-1? min-1, t=4.449, P=0.001).And no significant difference in rCBF was found in migraine patients without aura under attack-free period. Conclusions Three-dimensional arterial spin labeling could quantitatively analyze the cerebral blood flow .Monitoring the perfusion difference in migraine patients could provide some references in the pathogenesis research and therapy.
8.Slow synaptic transmission mediated by 5-HT_(1P) receptor in sympathetic ganalion of guinea pig
Liecheng WANG ; Yan ZHU ; Yanlong HE ; Jinlan HU ; Zhenxin HUANG ; Ruchun MA
Chinese Pharmacological Bulletin 1986;0(05):-
AIM To investigate the effects of several 5-hydroxytryptamine(5-HT) subtype receptor antagonists on late slow excitatory postsynaptic potential(LS-EPSP) of the neurons of guinea pig inferior mesenteric ganglion(IMG). METHODS Intracellular recordings were made from neurons of the isolated guinea pig IMG. RESULTS Cyproheptadine and BRL 24924 suppressed LS-EPSP of 5-HT sensitive neurons reversibly, while mianserin, MDL 72222 and spiperone showed no significant effect. Continuous superfusion of IMG with MCPP suppressed LS-EPSP of 5-HT sensitive neurons by 5-HT 1P receptor desensitizated. CONCLUSION LS-EPSP of 5-HT sensitive neuron is mediated by 5-HT 1P subtype receptor.
9.Effects of Hydrogen Sulfide on Apoptosis of PC12 Cells Induced by Chemical Hypoxia
Jinlan MENG ; Aiping LAN ; Ruixian GUO ; Chuntao YANG ; Zhanli YANG ; Xue HUANG ; Jianqiang FENG
Journal of Sun Yat-sen University(Medical Sciences) 2010;31(1):79-84
[Objective] To explore the cytoprotecfion of hydrogen sulfide (H_2S) against cobalt chloride (CoCl_2)-induced apeptosis in PC12 cells and the underlying mechanisms. [Methods] CoCl_2 (a chemical hypoxia-mimetic agent) was used to establish the chemical hypoxia-induced PC12 cell injuries model. Sodium hydrosulfide (NaHS) was used as a H_2S donor. The viability of PC12 cells was measured by CCK-8 assay. The percentage of apeptotic cells was assessed by propidium iodide stain flow cytometry (FCM). The morphological change of apeptotic cells was tested by using the chromatin dye Hoechst 33258. The mitochondrial membrane potential (MMP) was analyzed by rhodamine 123 staining and photofluorography. The level of reactive oxygen species (ROS) in PC12 cells was measured by DCFH-DA staining and photofluorography. [Results] CoCl_2 induced a decrease in cell viability and an increase in percentage of apeptosis in PC12 cells along with dissipation of MMP as well as overproduction of ROS. When PC12 cells were treated with Naris 30 min before CoCl_2 treatment a decrease in viability of PC12 cells induced by 600 μmol/L CoCl_2 was concentration-dependently blocked by NaHs (100, 200, and 400 μmol/L). Pretreatment with NaHS at 200 and 400 μmol/L obviously reduced the apepetotic percentage of PC12 cells induced by 600 μmol/L CoCl_2 and inhibited the dissipation of MMP and overproduction of ROS. [Conclusion] H_2S protected PC12 cells against CoCl_2-induced apeptosis, which may be associated with the inhibition of H_2S on the dissipation of MMP and overproduction of ROS induced by CoCl_2.
10.Study of clinical intervention of Shanzha Xiaozhi capsule on obesity patients with dampness-heat constitution
Jinlan LAI ; Qijun LIANG ; Rong HUANG ; Ruizhu CHEN ; Chenming HU ; Shouyi YU
Tianjin Medical Journal 2017;45(9):940-943
Objective To investigate the effects of Shanzha Xiaozhi capsule on body weight, metabolic index, percentage and distribution of body fat in obese patients with dampness-heat constitution. Methods A total of 60 obesity patients with dampness-heat constitution were enrolled in this study and randomly categorized into the control group ( n=30) and treatment group (n=30). Control group was given the diet and exercise regime, and treatment group was given Shanzha Xiaozhi capsule based on the same diet and exercise regime for 12 weeks. Scores of dampness-heat, body weight, waist circumference, hip circumference, blood pressure, blood glucose, serum lipid profile, urine acid (UA), fasting plasma insulin, serum levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-6, adiponectin, lipopolysaccharide (LPS), percentage and distribution of body fat were measured before and after treatment. Results (1) Data of scores of dampness-heat and diastolic blood pressure were significantly decreased after treatment in drug group. The above indexes and hip circumference were significantly decreased after the treatment in treatment group compared with those of control group ( P<0.05). (2) The triglyceride (TG), UA, insulin, HOMA-IR, LPS and IL-1βwere significantly decreased after the treatment in treatment group than those of control group, while there were no significant differences in fast blood glucose (FBG), 2 h postprandial plasma glucose (2 hPG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), TNF-α, IL-6 and adiponectin between two groups. (3) The percentage of visceral fat was significantly decreased in treatment group than that of control group. There were no significant differences in fat changes of limbs and buttocks between two groups. Conclusion Shanzha Xiaozhi capsule can decrease body weight and visceral fat content, reduce insulin resistance, and improve chronic inflammation state in obese patients with dampness- heat constitution.