1.Evaluation of hot-flash model rats induced by bilateral oophorectomy and tamoxifen administration
Haina XIE ; Zhiqiang PAN ; Linna CAO
Acta Laboratorium Animalis Scientia Sinica 2023;31(12):1545-1554
Objective To explore the animal model of syndrome used in the study of hot flash phenomenon in women.Methods Twenty-four female SD rats were divided randomly into three groups:Con group,Ovx group,and tamoxifen group(n = 8 rats per group).Hot flashes were induced by bilateral oophorectomy and intragastric tamoxifen 10 mg/(kg·d),respectively.Open-field activity,anal temperature,and body surface infrared thermograms were detected on model days 14 and 28.The rats were then killed on day 29 and their uteruses were removed,weighed,and sectioned.Blood estradiol and catecholamine levels were determined by enzyme-linked immunosorbent assay.Gene expression levels of adrenal sex hormone synthetases(Star,Cyp11a1,Cyp17a1,Cyp19a1,Por,Hsd3b2,Hsd17b1)and catecholamine synthetases(Th,Ddc,Dbh,Pnmt)in the adrenal medulla were detected by reverse transcription-polymerase chain reaction.Results Rat body weight was significantly higher in the Ovx group compared with Con group(P<0.01),while body weight increased slowly in the tamoxifen group.The maximum body surface temperature was significantly decreased on day 28 in the Ovx group(P<0.01),the difference between the maximum and minimum abdominal temperatures was significantly increased on day 14(P<0.05),the difference between the maximum and minimum temperatures on the back was significantly increased on day 28(P<0.01),and the open-field activity was decreased(P<0.01).Compared with the sham operation group,the maximum body surface temperature in the tamoxifen group was significantly decreased(P<0.01)but the open-field activity was increased(P<0.01).The uterine index was significantly decreased in both models(P<0.01).Estradiol levels were significantly decreased(P<0.01)and NE and epinephrine were also significantly decreased in the Ovx group compared with Con group(P<0.05),and β-EP was also significantly decreased in Ovx group(P<0.05).Adrenal Cyp11a1 gene expression was significantly increased(P<0.05)while Cyp17a1 and Hsd17b1 gene expression levels were significantly decreased(P<0.05)in bilateral ovariectomized rats compared with Con group.Compared with Con group,gene expression levels of Star and Por were significantly increased(P<0.01)while Cyp17a1 gene expression was significantly decreased(P<0.01)in the tamoxifen group,and Pnmt gene expression was significantly down-regulated in Ovx group(P<0.01).Conclusions Bilateral ovariectomized rats can be used for the study of perimenopausal hot flashes,which resemble kidney Yang and Yin deficiency in traditional Chinese medicine.
2.The short-term efficacy and safety of HA280 immunoadsorption column in patients with idiopathic inflammatory myopathies
Shu LI ; Haina GAN ; Suqing XU ; Jinfeng DU ; Fen LI ; Xi XIE ; Jia WANG ; Jinwei CHEN
Chinese Journal of Rheumatology 2020;24(4):247-252
Objective:To evaluate the short-term efficacy and safety of HA280 immunoadsorption (IA) column in idiopathic inflammatory myopathies (IIM).Methods:The clinical data of 72 patients with IIM admitted to the Department of Rheumatology of Xiangya No.2 Hospital of Central South University from January 2015 to March 2018 were analyzed. Of these patients, 22 patients were treated with HA280 immunoadsorption column for three times (the immunoadsorption group) and 50 patients were treated with drugs only (the control group). The changes of clinical symptoms and signs, autoimmune antibodies, myocardial enzyme spectrum, the inflammatory markers [erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) and ferritin], immunoglobulin, complement, other biochemical indexes and pulmonary images of the patients were detected and analyzed before and after the treatment. And then the data were analyzed by Chi-square test, samples t testand Wilcoxon rank sum test. Results:Compared with the control group, the symptoms and signs were obviously improved after treatment with HA280 immunoadsorption column. In particular, the clinical improvement rate of non-specific myositis (89%, 16/18) was higher than that of the control group [(58%, 22/38), χ2=5.379, P<0.05]. And the clearance of autoantibody (control group) was grade 39.41 in average, 28.38 in average in the immunoadsorption group( Z=-2.51, P=0.01), myocardial enzyme spectrum [control 717(1 564) U/L, immunoadsorption group 126(432) U/L , Z=3.09, P<0.01], the inflammatory markers such as ESR [the control group was 24(22) mm/1 h, the immunoadsorption group was 10(7) mm/1 h, Z=-3.0, P=0.003] and immunoglobulin G [the control group was 11(5) g/L, the immunoadsorption group was 9(2) g/L, Z=-4.8, P=0.001] and immunoglobulin M [the control group was 0.9(0.4) g/L, the immunoadsorption group was 1.2(0.8) g/L, Z=-2.0, P=0.05]. Moreover, the lung CT scan showed that pulmonary lesions of the patients in the immunoadsorption group (89%, 17/19) was much more improved than the control group [(61%, 27/44), χ2=4.98, P<0.05]. No serious adverse reactions occurred. Conclusion:HA280 immunoadsorption therapy can significantly clear the autoantibodies, decrease muscle enzymes, inflammatory markers and immunoglobulin, improve lung images of some patients in a short time. It has been shown that it is safein patients with IIM. HA280 immunoadsorption therapy could be an effective treatment for IIM.
3.Expression of death receptor 5 upreguleted by chloroquine enhances the sensitivity of Huh7 cells to tumor necrosis factor related apoptosis-inducing ligand
Yanfeng WANG ; Liwu XIE ; Hongxia WANG ; Wenwei YAN ; Haina WANG ; Li JIA ; Peng BU ; Haixia MA ; Yanfeng XI
Cancer Research and Clinic 2018;30(7):438-442
Objective To explore the effect of chloroquine on death receptor 5 (DR5) expression of hepatocellular carcinoma Huh7 cells and cell proliferation and apoptosis induced by tumor necrosis factor related apoptosis-inducing ligand (TRAIL).Methods Huh7 cells were divided into four groups:the control group (1∶1 000 dimethyl sulfoxide),TRAIL group (50 μg/L),chloroquine group (10 μmol/L) and TRAIL +chloroquine group (TRAIL 50 μg/L + chloroquine 10 μmol/L).Thiazolyl blue tetrazolium bromide (MTT) assay was used to determine the proliferation activity of cells,immunofluorescence was used to detect the expression of DR5,4',6-diamidino-2-phenylindole (DAPI) staining was used to observe cell apoptosis and Western blot was used to detect the expression of cleaved poly ADP-ribose polymerase (PARP).Results TRAIL treatment could decrease Huh7 cells proliferation activity;when compared with the cell viability in the control group,the cell proliferation inhibition rate of chloroquine group,TRAIL group and TRAIL+ chloroquine group was (89±8) %,(53±10) % and (27±7) %,respectively;compared with TRAIL group alone,cell proliferation activity was decreased in TRAIL+ chloroquine group (t =3.922,P =0.017).The expression of DR5 was upregulated in chloroquine group,and the cell apoptosis signaling was activated in TRAIL + chloroquine group.The cell apoptosis rate of TRAIL group and TRAIL + chloroquine group was (10.0±2.3) % and (20.4±4.0) %,respectively,and there was a statistical difference (t =3.894,P =0.018).Conclusion Chloroquine can enhance the cell chemosensitivity to TRAIL treatment by upregulating the expression of DR5 in Huh7 cells.
4.Injurious Effect of Cisplatin on the Function of Hypothalamus-pituitary-adrenal/gonadal Axis in Mice and the Intervention Effect of Dehydroepiandrosterone
Zhiqiang PAN ; Zixin NONG ; Haina XIE ; Peike PENG
Laboratory Animal and Comparative Medicine 2023;43(3):229-242
ObjectiveTo study the pathway of cis-dichlorodiamineplatinum (DDP) inhibiting the synthesis of steroid hormones in mice, and to observe the intervention effect of dehydroepiandrosterone (DHEA).MethodsSixty adult ICR mice were randomly divided into three groups: control group, DDP modeling group, and DHEA group, with 10 male and 10 female mice in each group. The DDP modeling group mice were intraperitoneally injected with DDP solution at a dose of 2.5 mg·kg-1·d-1, once every 3 days, a total of 7 times. On the same day of modeling, the control group mice were injected with an equal amount of physiological saline intraperitoneally. The DEHA treatment group mice were treated with DDP and given a dose of 8.3 mg·kg-1·d -1 of DHEA by gavage for 21 consecutive days. The changes of fatigue indexes of mice were observed by open field, grip and rod rotation tests. The morphology changes of adrenal gland, testicular and ovarian tissue were observed by pathological section and HE staining. The levels of serum steroid hormones were detected by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The mRNA and protein expression levels of the related genes of the hypothalamus, hypophysis, adrenal, testis and ovary were tested by real-time fluorescent quantitative PCR (RT-qPCR) and Western blotting.ResultsCompared with control group, both male and female mice in DDP modeling group were significantly losing weight (P<0.05), their abilities in horizontal movement and vertical movement decreased (all P<0.05), and the stay time and grip also significantly decreased (all P<0.05) in female mice. Indexes of fatigue were improved after DHEA supplement (all P<0.05). In the DDP modeling group, the arrangement of spermatogenic cells at all levels in the testicular tissue was disordered and the testicular interstitial edema was observed, and a large number of primordial follicles in the ovarian tissue were activated, the number of atresia follicles increased, and the number of granulosa cells in the follicles decreased; while in the DHEA group, the damaged phenotype of testicles and ovaries was significantly improved. Compared with control group, the levels of serum testosterone and dihydrotestosterone in both male and female DDP modeling mice significantly decreased (P<0.01), the pregnenolone was down-regulated but corticosterone was up-regulated significantly (P<0.05) in male mice, the corticosterone was down-regulated significantly (P<0.05) in female mice. Compared with the DDP group, after DHEA supplement, the pregnenolone in male mice and the progesterone in female mice increased significantly (P<0.05), but the pregnenolone in female mice and the progesterone in male mice decreased significantly (P<0.05). Compared with control group, the expression levels of Cyp21a1 and Cyp11a1 genes in the adrenal gland and Gnrh gene in the hypothalamus of male and female mice in the DDP modeling group significantly decreased (all P<0.05); the expression levels of Hsd3b2 gene in the adrenal gland, Star, Cyp11a1, and Lhr genes in the ovaries, Crh, Pomc, and Lhb genes in the hypothalamus, pituitary, and pituitary of female mice significantly decreased (all P<0.05); the expression levels of Star gene and StAR protein in the testicles of male mice, as well as Fshb and Lhb genes in the pituitary gland, were significantly down-regulated (all P<0.05). After DHEA supplement, compared with the DDP modeling group, the mRNA expression levels of Cyp17a1 in the adrenal gland of male mice and Cyp17a1, Lhr and Fshr genes in testis were down-regulated significantly (P<0.05); the expression level of Cyp11a1 gene in the adrenal gland of female mice was also decreased (P<0.05); while the expression levels of Hsd3b2 gene in the adrenal gland, Star, Cyp11a1, Hsd3b2 and Lhr gene in the ovary, and Lhb gene in the pituitary gland were all up-regulated ( P<0.05).ConclusionThe function of hypothalamus-pituitary-adrenal/gonadal axis was inhibited by DDP intermittent injection, especially in female. Supplementation of DHEA can help regulate the homeostasis of steroid hormone levels.