1.Hair Growth Promoting Effects of 650 nm Red Light Stimulation on Human Hair Follicles and Study of Its Mechanisms via RNA Sequencing Transcriptome Analysis
Kai YANG ; Yulong TANG ; Yanyun MA ; Qingmei LIU ; Yan HUANG ; Yuting ZHANG ; Xiangguang SHI ; Li ZHANG ; Yue ZHANG ; Ji’an WANG ; Yifei ZHU ; Wei LIU ; Yimei TAN ; Jinran LIN ; Wenyu WU
Annals of Dermatology 2021;33(6):553-561
Background:
Androgenetic alopecia (AGA) leads to thinning of scalp hair and affects 60%~70% of the adult population worldwide. Developing more effective treatments and studying its mechanism are of great significance. Previous clinical studies have revealed that hair growth is stimulated by 650-nm red light.
Objective:
This study aimed to explore the effect and mechanism of 650-nm red light on the treatment of AGA by using ex vivo hair follicle culture.
Methods:
Human hair follicles were obtained from hair transplant patients with AGA. Hair follicles were cultured in Williams E medium and treated with or without 650-nm red light.Real-time RT-PCR and immunofluorescence staining were used to detect the expression level of genes and proteins in hair follicles, respectively. RNA-sequencing analysis was carried out to reveal the distinct gene signatures upon 650 nm treatment.
Results:
Low-level 650 nm red light promoted the proliferation of human hair follicles in the experimental cultured-tissue model. Consistently, 650 nm red light significantly delayed the transition of hair cycle from anagen to catagen in vitro. RNA-seq analysis and gene clustering for the differentially expressed genes suggests that leukocyte transendothelial migration, metabolism, adherens junction and other biological process maybe involved in stimulation of hair follicles by 650-nm red light treatment.
Conclusion
The effect of 650-nm red light on ex vivo hair follicles and the transcriptome set which implicates the role of red light in promoting hair growth and reversing of miniaturization process of AGA were identified.
2.Comparison of Anti-hepatocarcinoma Effect of Curcumin and Hydrazincurcumin and Mechanism Study
Ji’an ZHAO ; Limin CUI ; Liang DONG ; Wenjia NIE ; Wencong LIU ; Zengning LI
China Pharmacy 2020;31(22):2741-2750
OBJECTIVE:To compare the an ti-hepatocarcinoma effects of curcumin (CUR)and its derivative hydrazincurcumin (HZC),and to explore the mechanism. METHODS :MTT assay was used to detect the effects of CUR or HZC (2.5,5,10,20, 40,80 μmol/L)on the proliferation of HepG 2 cells. Flow cytometry was used to detect the effects of CUR or HZC (10,20,40 μmol/L)on cell cycle distribution and apoptosis of HepG 2 cells. Western blotting assay was used to detect the effects of CUR or HZC(10,20,40 μmol/L)on the expression of apoptosis-related protein in HepG 2 cells. The male SD rats were randomly divided into normal control group (n=10),CUR control group (n=10),HZC control group (n=10),model group (n=30),CUR protection group (n=30)and HZC protection group (n=30). CUR control group and HZC control group were given CUR 85917439。E-mail:zhaoji-an-88@163.com or HZC (80 mg/kg) intraperitoneally. Model group ,CUR protection group and HZC protection group were given diethylnitrosamine (50 mg/kg)intraperitoneally to establish hepatocarcinoma model ;at the same time ,2 protection groups were given CUR or HZC (80 mg/kg)intraperitoneally,twice a day,for consecutive 12 weeks. During medication ,the change of body weight and death of rats were recorded. Twenty four weeks later,liver index of rats was calculated and appearance was observed ;the number of cancer nodules was counted ;HE staining was used to observe the pathological changes of liver tissue and calculate the nuclear division index of hepatocarcinoma ;the proliferating cell nuclear antigen (PCNA)index was detected by immunohistochemistry. RESULTS :CUR and HZC could increase the inhibitory rate of HepG 2 cells(P<0.05),and increased the percentage at G 0/G1 phase and apoptotic rate of HepG 2 cells(P< 0.05). CUR and HZC could significantly decrease the protein expression of p-JAK 2,p-STAT3,Bcl-2 and Bcl-xl ,while increased the protein expression of Bax ,Cyt-c,Caspase-9,Caspase-3 and PAPR (P<0.05). Above effects of HZC were significantly better than those of CUR (P<0.05). The results of animal experiment showed that there was no death ,no liver canceration and no pathological changes in liver appearance and tissue section of the three control groups ;there was no statistical significance in body weight and its increased weight ,liver index ,nuclear division index of carcinoma or PCNA index (P>0.05). Compared with model group, survival rate of rats were increased significantly in CUR protection group and HZC protection group , while hepatocarcinoma incidence and the number of cancer nodules were decreased significantly (P<0.05);body weight and its increased weight were increased significantly ,while liver index ,nuclear division index of carcinoma and PCNA index were decreased significantly (P<0.05). There were some pathological changes in liver appearance and tissue section ;cancerous lesions with focal necrosis or cancerous lesions with patchy necrosis were observed. There was no statistical significance in the improvement of above indexes in 2 protection groups (P>0.05). CONCLUSIONS :HZC could inhibit the proliferation and induce apoptosis of HepG 2 cells by inhibiting JAK 2/STAT3 signaling pathway and regulating the activation of mitochondrial endogenous pathway,which shows stronger anti-hepatocarcinoma effect in vitro than CUR. On the other hand ,there was no significant difference in the improvement of liver caner indexes in hepatic cancer model rats between HZC and CUR.
3.Effect and mechanism of gracillin-induced autophagy in lung cancer A549 cells
Yan LI ; Yamei LI ; Geyan LEI ; Jialan KANG ; Mingxuan LIU ; Minhong ZHANG ; Jianqiong YANG
China Pharmacy 2024;35(8):912-917
OBJECTIVE To investigate the effect and mechanism of gracillin from Reineckia carnea on autophagy in non- small cell lung cancer A549 cells. METHODS Using A549 cells as subjects, the effects of different concentrations of gracillin (0.25, 0.5, 1, 2, 4 μmol/L) on the proliferation of cells were detected by CCK-8 after being treated for different time (12, 24, 48 h). Compared with the control group without medication, the effect of gracillin (2 μmol/L) on the formation of autophagosomes in cells was observed by transmission electron microscope after 24 h of exposure. The aggregation of GFP-LC3 on autophagosome membrane was detected by GFP-LC3 plasmid transfection after being treated with gracillin (0.25, 0.5, 1, 2 μmol/L) for 24 h. Quantitative real-time PCR and Western blot assay were used to detect the mRNA and protein expressions of family with sequence similarity 102 member A(FAM102A), the expressions of autophagy-related proteins [p62, Beclin-1, microtubule-associated protein 1 light chain 3B (LC3B)], and the expressions of phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway-related proteins in A549 cells after being treated with gracillin (0.25, 0.5, 1 and 2 μmol/L) for 24 h. RESULTS Gracillin significantly inhibited the proliferation of A549 cells in a concentration- and time-dependent manner. The IC50 was 2.55 μmol/L at 24 h. After 24 h of gracillin treatment, autophagosomes with bilayer membrane structure were found in the cell cytoplasm, and GFP-LC3 green fluorescent spots on autophagosome membrane were obvious, representing an increasing trend as drug concentration. Compared with the control group, mRNA and protein expressions of FAM102A (0.5, 1, 2 μmol/L groups), protein expression of Beclin-1 (1, 2 μmol/L groups) and LC3B-Ⅱ/LC3B-Ⅰ ratio (2 μmol/L group) were significantly increased in different concentrations of gracillin groups, while the protein expression of p62 (1, 2 μmol/L groups), and the protein phosphorylations of Akt (1, 2 μmol/L groups) and PI3K (2 μmol/L group) were all decreased significantly (P<0.05 or P<0.01). CONCLUSIONS Gracillin can promote excessive autophagy in A549 cells by up-regulating mRNA and protein expressions of FAM102A and inhibiting PI3K/Akt signaling pathway, thus inhibiting cell proliferation.
4.Study on Improvement Effects and Mechanism of Paeoniflorin on Myocardial Injury in Type 2 Diabetic Model Rats
Shanshan LI ; Chunyu TIAN ; Guowei ZHANG ; Leilei MA ; Ji’an LI ; Jiaxuan LIU ; Ying LIU ; Shuang LI ; Xiaolong LI ; Jiahuan SHEN
China Pharmacy 2021;32(23):2846-2853
OBJECTIVE:To study the improvement effects of paeon iflorin(PF)on myocardial injury in type 2 diabetes mellitus(T2DM)model rats and its mechanism. METHODS :The experiment was set up in the normal group ,model group , positive control group (metformin 90 mg/kg),PF high-dose ,medium-dose and low-dose groups (90,60,30 mg/kg),with 8 rats in each group. Except for normal group ,other groups were given high-glucose and high-fat diet and intraperitoneal injection of streptozotocin (30 mg/kg) to induce T 2DM model. After modeling , administration groups were given relevant medicine intragastrically,normal group and model group were given constant volume of normal saline intragastrically ,once a day ,for consecutive 4 weeks. The body weight ,fasting blood glucose and oral glucose tolerance were measured ;serum levels of glycosylated serum protein (GSP),total cholesterol (TC),triacylglycerol(TG),glutathione peroxidase (GSH-Px),superoxide dismutase(SOD),malondialdehyde(MDA),creatine kinase isoenzyme-MB (CK-MB) and troponin Ⅰ (cTn Ⅰ) were determined. The pathomorphological changes of myocardium were observed. The apoptosis index of rat cardiomyocytes was ( detected. The protein expression of B-cell lymphoma 2 (Bcl-2),Bcl-2 related X protein (Bax)and caspase- 3 in rat myocardium were detected by immunohistochemistry and Western blot. RE SULTS:Compared with normal group ,the body weight ,serum levels of GSH-Px and SOD ,protein expression of Bcl- 2 in myocardium were decreased significantly in model group(P<0.01);while fasting blood glucose ,area under blood glucose curve ,serum levels of biochemical indexes (GSP,TC, TG,MDA,CK-MB,cTnⅠ),cardiomyocyte apoptosis index ,protein expression of Bax and caspase- 3 in myocardium were increased significantly (P<0.05 or P<0.01). The arrangement of myocardium was relatively irregular ,and some muscle fibers were broken. Compared with model group ,except for body weight ,serum levels of SOD and MDA ,the protein expression of Bax in myocardium in PF low-dose group , above indexes of PF groups were reversed significantly (P<0.05 or P<0.01). CONCLUSIONS:PF can regulate glycolipid metabolism ,enhance antioxidant ability ,inhibit cardiomyocyte apoptosis and improve myocardial injury in T 2DM model rats ;the mechanism may be associated with increasing the protein expression of Bcl- 2 and down-regulating the protein expression of Bax and caspase- 3 in myocardium.