1.Effects of astragaloside on TRPC6 expression on mouse podocyte induced by TGF-β1
Haiting HUANG ; Haohao WU ; Youling QIN ; Xu LIN ; Yanwu YOU ; Pengwei GUO ; Chunrong TANG
Chinese Journal of Immunology 2017;33(3):370-373
Objective:To explore the possible mechanism of astragaloside involved in the mouse podocytes injury induced by TGF-β1 in vitro.Methods:Mouse podocytes were cultured in vitro and then all cell were divided into 5 groups:normal control group , TGF-β1 treatment group ,TGF-β1 treatment +astragaloside low dose group ,TGF-β1 treatment +astragaloside middle dose group and TGF-β1 treatment +astragaloside high dose group.The proliferation rate of each group was investigated by MTT assay ,the expression of TRPC6 protein and mRNA were measured by Western blot and RT-PCR respectively after 48 hours.Results:TGF-β1 can significantly inhibit the proliferation of podocytes ( P<0.05) ,fusions of foot processes or even effaced of podocytes were observed .TGF-β1 could also increase the expression of TRPC6.Astragaloside could reduce the inhibition of TGF-β1 to the proliferain of podocytes significantly ,make the cell shape tend to be normal,and reduce the expression of TRPC6 mRNA and protein with dose-effect relation.Conclusion:TRPC6 play an impor-tant role in the TGF-β1 induecd podocytes injury .Astragaloside can alleviate podocytes injury by reduce the expression of TRPC 6.
2.Role of transient receptor potential cation channel 6 (TRPC6) channel in mouse podocyte injury induced by TGF-β1
Haiting HUANG ; Xu LIN ; Yanwu YOU ; Chunrong TANG ; Xianjun GU ; Meiying HUANG ; Junhua TAN ; Youling QIN ; Feifan HUANG
Chinese Journal of Microbiology and Immunology 2017;37(5):340-346
Objective To investigate the effects of transient receptor potential cation channel 6 (TRPC6) on the expression of nephrin, desmin and caspase 9 and on the apoptosis of podocytes in a mouse model of podocyte injury induced by TGF-β1.Methods Conditionally immortalized mouse podocytes were cultured in vitro and divided into four groups: control, TGF-β1 treatment, TGF-β1+PGPU6/GFP/Neo-TRPC6-mus-581 (TRPC6 knockdown) and TGF-β1+PGPU6/GFP/Neo-NC (negative control).Real-time RT-PCR and Western blot analysis were performed to detect the expression of nephrin, desmin and caspase 9 at mRNA and protein levels, respectively.Flow cytometry was used to analyze the apoptotic rate of podocytes.DAPI fluorescent staining was used to observe the morphological changes of apoptotic podocytes.Results Green fluorescent protein (GFP)-expressing podocytes at 48 hours after transfection were significantly more than those at 24 hours after transfection.The level of TRPC6 in mouse podocytes transfected with PGPU6/GFP/Neo-TRPC6-mus-581 was significantly decreased as compared with that of the control group (P<0.05).No significant difference in the expression of TRPC6 was observed between the negative control group and the control group.Compared with the control group, the TGF-β1 treatment group showed increased expression of desmin and caspase 9 at both mRNA and protein levels (P<0.01), but decreased expression of nephrin at mRNA and protein levels at 48 hours after TGF-β1 intervention (P<0.05).The up-regulated desmin and caspase 9 and the down-regulated nephrin induced by TGF-β1 could be inhibited by the means of TRPC6 knockdown.The apoptosis rate of podocytes in TGF-β1 treatment group was (14.0±2.1)%, while that in TRPC6 knockdown was (10.90±0.56)% (P<0.05).The apoptosis rate of podocytes in negative control group was higher than that in TGF-β1 treatment group (P>0.05).More apoptotic cells with typical morphological features of apoptosis were observed after exposure to TGF-β1 for 48 hours.Conclusion TGF-β1 could induce the apoptosis of podocytes, inhibit the expression of nephrin and enhance the expression of caspase 9 and desmin, the possible mechanisms of which may be related to TRPC6 signal pathway.These changes in TGF-β1-treated podocytes could be alleviated by inhibiting the expression of TRPC6, which might have a protective effect on podocyte injury.
3.Questionnaire investigation of the awareness of doctors' attitude to geriatric syndrome in old patients with cancer
Lingxiao WANG ; Zhangmin MENG ; Shuangshuang NIE ; Bing XIANG ; Jun LI ; Qianqian SUN ; Hai QIN ; Youling GONG ; Changchuan PAN ; Cheng YI ; Yongxue YANG ; Shuang WANG
Chinese Journal of Geriatrics 2017;36(5):579-582
Objective To improve the quality of the management of the elderly patients with cancer in Sichuan province,and to understand the attitudes and perspectives of elderly surgeons and oncologists for the management and treatment of elderly patients with cancer.Methods A face-toface questionnaire interview was conducted with oncologists (n 64) and geriatricians (n =64).128 physicians were involved in this study.Results The cancer management and therapeutics were deemed appropriate at present by 9.38% (6/64)of the geriatricians and 25.00% (16/64)of the oncologists.The 39.06% (25/64) of geriatricians used to notice geriatric syndromes,while 81.25% (52/64)of oncologists never concerned about the geriatric syndrome(P=0.011).As for the causes of the therapy-associated toxicity,oncologist versus geriatrician payed an attention to malnutrition (100.00% vs.100.00% in both groups),to mobility disorders(65.63% vs.65.63%,84/128 in both groups),to cognitive impairment/mood disorder (89.06% in geriatrician group vs.75.00% in oncologist group,P=0.038).For the factors affecting treatment decisions,a physical ability attention (oncologists vs geriatricians:70.31% vs.92.19%;P =0.002),and comorbidity (oncologists vs geriatricians:62.50% vs.79.69%,P =0.032) had statistically significant difference.In addition,lack of geriatrics knowledge was also reported by more oncologists.However,one hundred percent of participants wanted very much to cooperate with each other in their clinical work.When responding to the clinical scenario,the 10.94% (7/64)of geriatricians and 32.81% (21/64)of oncologists chose modified treatment for 65-74 years old patient with cancer(P =0.003).When the age of the patients was 75-84 years old,only 12.50 % (8/64) of geriatricians prefer end-of-life care,while 31.25 % (20/64) of oncologists chose it (P =0.010).Conclusions Selection of treatment decisions in the elderly patients with cancer affect by ageing.Both oncologists and geriatricians are concerned with the elderly patients with cancer,ageing syndrome,total sickness and functional status.And these doctors support an establishment of a multi disciplinary team cooperation for the elderly patients with cancer.Therefore,the establishment of mutual cooperation between the two professionals is necessary and feasible.