CN106771242B - A kind of experimental method proving hydrogen sulfide and gap connection relationship - Google Patents
A kind of experimental method proving hydrogen sulfide and gap connection relationship Download PDFInfo
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- CN106771242B CN106771242B CN201611169906.1A CN201611169906A CN106771242B CN 106771242 B CN106771242 B CN 106771242B CN 201611169906 A CN201611169906 A CN 201611169906A CN 106771242 B CN106771242 B CN 106771242B
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Abstract
The invention discloses a kind of experimental methods for proving hydrogen sulfide and gap connection relationship, method includes the following steps: animal blood pressure measures；The preparation of Peripheral Lymphocytes of Rat suspension；The detection of T lymphocyte subsets and Cx40, Cx43.Present invention firstly discloses the relationships that albumen is connected on hydrogen sulfide and T lymphocyte, hydrogen sulfide is prompted to pass through the gap connecting communication between influencing immunocyte, the release for adjusting inflammatory factor, to play important function in improving target organ damage caused by hypertension inflammation.
Description
Technical field
The invention belongs to field of biotechnology, specifically, being related to a kind of reality for proving hydrogen sulfide and gap connection relationship
Proved recipe method.
Background technique
Peripheral blood lymphocytes is divided into 3 groups: T lymphocyte (CD3 according to biological function and cell surface antigen expression
+), bone-marrow-derived lymphocyte (CD19+) and NK lymphocyte [natural killer cells (natural killer), CD3-CD16+ and/or
CD56+].T cell is divided into T helper cell (CD3+CD4+) again and inhibits T cell (CD3+CD8+).Lymphocyte subgroup is analyzed
Detect cellular immunity and humoral immune function important indicator, the current immune function of its general reaction body, state and
Equilibrium level, and can be with the certain diseases of auxiliary diagnosis, such as autoimmunity disease, immunologic deficiency disease, malignant tumour, blood disease, metamorphosis
Reactive disorder etc. analyzes pathogenesis, observes curative effect and detection prognosis is significant.Clinically find gap junction protein
40 gene C x40 (- 44 and+71 site) polymorphism has certain correlation with essential hypertension (EH).
Currently, in the prior art, there are no the relevant reports for the experimental method for proving hydrogen sulfide and gap connection relationship.
Summary of the invention
It is an object of the invention to overcome defect existing for above-mentioned technology, a kind of easy to operate, low-cost card is provided
The experimental method of bright hydrogen sulfide and gap connection relationship.
Itself the specific technical proposal is:
A kind of experimental method proving hydrogen sulfide and gap connection relationship, comprising the following steps:
Step 1, animal blood pressure measurement
Three groups of WKY rat, SHR and hydrogen sulfide intervention group big rat-tails are detected using full-automatic rats and mice non-invasive blood pressure measuring instrument
Arterial systolic blood pressure.It is first preheated when measurement, the inflation tail sleeve of sphygmomanometer is covered in rat root of the tail portion, after rat is quiet, repeat to survey
It takes blood pressure 3 times, as a result takes its average value.
Step 2, the preparation of Peripheral Lymphocytes of Rat suspension
By under rat anesthesia state, abdominal aorta peripheral blood is acquired, is placed in 10g/LEDTA vacuum blood collection tube.Peripheral blood
It is centrifuged, serum and haemocyte is separated, few one times of PBS solution is added in haemocyte, is mixed well, after dilution
Haemocyte be gently laid on equivalent lymphocyte separation medium, 1500r, 30min, extract cloud in 1.5ml centrifuge tube,
1800r, 6min abandon supernatant, and PBS solution is resuspended, spare.
The detection of step 3, T lymphocyte subsets and Cx40, Cx43
Streaming special dyeing pipe is taken, each group rat is respectively labeled as blank, measurement pipe and Isotype control pipe in sequence, respectively
Pipe is separately added into corresponding streaming antibody, adjusts antibody final concentration according to antibody specification, and set up negative control.Room temperature, which is protected from light, incubates
30min is educated, CELLQuest software is used on the machine and is detected.It marks and 120 μ of fixative is added in the pipe of CD4-APC and CD8-PE
L, 4 DEG C of incubation 30min, then with rupture of membranes buffer rupture of membranes, according to specification reference concentration be added goat anti-rat Cx40 antibody and
The goat anti-mouse of the rabbit-anti goat IgG of FITC label, FITC Isotype control and small anti-rat Cx43 antibody and FITC label
IgG, FITC Isotype control, 37 DEG C of incubation 30min, use on the machine CELLQuest software and are detected.Flow cytometer is adjusted
To optimum Working, with FSC/SSC gating and dead cell and cell fragment are excluded, experiment parameter is adjusted according to Isotype control
Carry out sample measures again afterwards, gating first is in CD3+ lymphocyte, then lymph of the gating after CD4+ and CD8+ cell, rupture of membranes
Cell gating is in CD4+ and CD8+, and then for gating in Cx40+, Cx43+ cell, all sample standard deviations complete the inspection to cell in 2h
It surveys, every pipe detects 20000 cells, and computer reads measured value, finally analyzes the data of collection.
Further, corresponding streaming antibody specific described in step 3 are as follows: CD3-FITC, FITC Isotype control；CD4-APC,
APC Isotype control；CD8-PE, PE Isotype control；The rabbit-anti goat IgG of goat anti-rat Cx40 antibody and FITC label, FITC
Isotype control；The goat anti-mouse IgG of small anti-rat Cx43 antibody and FITC label, FITC Isotype control.
Compared with prior art, beneficial effects of the present invention: present invention firstly discloses connect on hydrogen sulfide and T lymphocyte
The relationship of albumen is connect, prompts hydrogen sulfide by the gap connecting communication between influencing immunocyte, adjusts the release of inflammatory factor, from
And important function has been played in improving target organ damage caused by hypertension inflammation.
Detailed description of the invention
Fig. 1 is the flow chart present invention demonstrates that the experimental method of hydrogen sulfide and gap connection relationship；
Fig. 2 is the comparison of three groups of rat artery blood pressures；
Fig. 3 is three groups of rat kidney tissue pathologies changes (× 400)；
Fig. 4 is three groups of rat peripheral blood t lymphocyte subset group frequencys；Wherein, Fig. 4 A:CD3+ expression frequency；Fig. 4 B:CD8+
Expression frequency；Fig. 4 C:CD4+ expression frequency；Fig. 4 D:CD4+CD25+ expression frequency
Fig. 5 is three groups of rat peripheral blood CD4+The expression of T Cx40 cell；
Fig. 6 is three groups of rat peripheral blood CD4+The expression of Cx43 cell；
Fig. 7 is three groups of rat peripheral blood CD8+The expression of T Cx40 cell；
Fig. 8 is three groups of rat peripheral blood CD8+The expression of T Cx43 cell.
Specific embodiment
Technical solution of the present invention is described in more detail with specific embodiment with reference to the accompanying drawing.
Grouping: WKY group: normal Wistar capital of a country (Wistar-Kyoto, WKY) rat, male and female are regardless of, 9 week old, daily abdomen
Chamber injection and SHR+H2S group same amount of normal saline, continue 8 weeks.
SHR group: spontaneous hypertensive rat (spontaneously hypertensive rat, SHR), male and female are regardless of, and 9
Week old, daily intraperitoneal injection and SHR+H2S group same amount of normal saline, continue 8 weeks.
NaHS processing group: SHR, male and female are regardless of, 9 week old, and the NaHS (H of 56 μm of ol/kg weight is injected intraperitoneally daily2The confession of S
Body), continue 8 weeks.
The measurement of 1 animal blood pressure detects each group rat tail artery systolic pressure using full-automatic rats and mice non-invasive blood pressure measuring instrument.
It is first preheated when measurement, the inflation tail sleeve of sphygmomanometer is covered in rat root of the tail portion, after rat is quiet, duplicate measurements blood pressure 3 times, knot
Fruit takes its average value.
The preparation of 2 Peripheral Lymphocytes of Rat suspensions acquires under rat anesthesia state abdominal aorta peripheral blood, is placed in
In 10g/LEDTA vacuum blood collection tube.Peripheral blood is centrifuged, and serum and haemocyte are separated, and few one is added in haemocyte
PBS solution again, mixes well, the haemocyte after dilution is gently laid on equivalent lymphocyte separation medium, 1500r, 30min,
Cloud is extracted in 1.5ml centrifuge tube, 1800r, 6min abandon supernatant, and PBS solution is resuspended, spare.
The detection of 3 T lymphocyte subsets and Cx40, Cx43 take streaming special dyeing pipe, and each group rat is according to suitable
Sequence is respectively labeled as blank, measurement pipe and Isotype control pipe, each pipe and is separately added into corresponding streaming antibody, according to antibody specification tune
Whole antibody final concentration, and set up negative control.Room temperature, which is protected from light, is incubated for 30min, uses on the machine CELLQuest software and is detected.
It marks and fixative 120 μ L, 4 DEG C of incubation 30min is added in the pipe of CD4-APC and CD8-PE, then with rupture of membranes buffer rupture of membranes, according to
The rabbit-anti goat IgG of goat anti-rat Cx40 antibody and FITC label, FITC Isotype control and small is added in specification reference concentration
The goat anti-mouse IgG of anti-rat Cx43 antibody and FITC label, FITC Isotype control, 37 DEG C of incubation 30min are used on the machine
CELLQuest software is detected.Flow cytometer is adjusted to optimum Working, with FSC/SSC gating and is excluded dead thin
Born of the same parents and cell fragment carry out sample measures after adjusting experiment parameter according to Isotype control again, and gating first is thin in CD3+ lymph
Born of the same parents, then lymphocyte gating of the gating after CD4+ and CD8+ cell, rupture of membranes be in CD4+ and CD8+, then gating in Cx40+,
Cx43+ cell, all sample standard deviations complete the detection to cell in 2h, and every pipe detects 20000 cells, and computer reads measurement
Value, finally analyzes the data of collection.
Experimental result:
(1) 8 week old that are compared to of WKY rat group, SHR group and NaHS intervention group rat blood pressure start to process each group rat,
After continuing 9 weeks, measure each group rat systolic pressure, the results showed that SHR group rat systolic pressure be significantly higher than WKY rat group (P <
0.01), NaHS intervention group rat systolic pressure is significantly higher than SHR group rat, Fig. 2.
(2) HE dyeing discovery, it is seen that WKY rat group Renal Glomeruli In Rats size rule, renal cells are complete, arrangement
Neatly, renal interstitial is without lesion；There is slight atrophy in SHR group renal blood vessels wall thickening, glomerulus, and vacuole occurs in Tubular epithelial cell
And granular degeneration, renal interstitial is by inflammatory cell infiltration；Atrophy of the visible glomerulus of NaHS intervention group compared to SHR group makes moderate progress,
The vacuole and granular degeneration of renal cells are less also relative to SHR group, and renal interstitial inflammatory infiltration degree is also relative to SHR
Less, the Fig. 3 of group.
(3) Flow cytometry WKY rat group, SHR group and NaHS intervention group rat peripheral blood lymphocyte subgroup
Expression frequency, the results show that the results show that WKY rat group, SHR group and NaHS intervention group peripheral blood CD4+T lymphocyte hundred
Divide than being (67.28 ± 1.31) %, (69.31 ± 0.98) % and (67.80 ± 0.38) %, no difference of science of statistics；WKY rat
Group, SHR group and NaHS intervention group peripheral blood CD8+T lymphocyte percentage be (34.62 ± 1.42) %, (33.40 ±
0.85) % and (34.63 ± 0.62) %, no difference of science of statistics；WKY rat group, SHR group and NaHS intervention group peripheral blood CD4+/
CD8+The ratio of T lymphocyte percentage is 2.01 ± 0.14,2.09 ± 0.08 and 1.96 ± 0.04, no difference of science of statistics；WKY
Rat group, SHR group and NaHS intervention group peripheral blood CD4+CD25+T lymphocyte percentage be (7.72 ± 0.50) %, (5.94 ±
0.33) % and (6.49 ± 0.35) %, SHR group vs WKY rat group reduce, and have statistical difference (P < 0.05), SHR group vs
NaHS intervention group increases, no difference of science of statistics, Fig. 4.
(4) WKY rat group, SHR group and NaHS intervention group rat peripheral blood T lymphocyte are measured by flow cytometry
The expression frequency of the upper Cx40 of CD4+ is respectively (5.20 ± 1.69) %, (18.60 ± 2.78) %, (2.90 ± 0.80) %, SHR group
It expresses and rises compared with WKY rat group, difference is statistically significant (P < 0.01), and NaHS intervention group is expressed compared with SHR group to be declined, and difference has
Statistical significance (P < 0.01), Fig. 5.
(5) WKY rat group, SHR group and NaHS intervention group rat peripheral blood T lymphocyte are measured by flow cytometry
The expression frequency of the upper Cx43 of CD4+ is respectively (0.21 ± 0.06) %, (0.25 ± 0.06) %, (0.23 ± 0.06) %, SHR group
It expresses and rises compared with WKY rat group, no significant difference, NaHS intervention group is expressed compared with SHR group to be declined, no statistical difference meaning
Justice, Fig. 6.
(6) WKY rat group, SHR group and NaHS intervention group rat peripheral blood T lymphocyte are measured by flow cytometry
The expression frequency of the upper Cx40 of CD8+ is respectively (1.98 ± 0.32) %, (0.85 ± 0.25) %, (1.29 ± 0.25) %, SHR group
Compared with WKY rat group express decline, difference is statistically significant (P < 0.01), NaHS intervention group compared with SHR group expression rising, difference without
Statistical significance, Fig. 7.
(7) WKY rat group, SHR group and NaHS intervention group rat peripheral blood T lymphocyte are measured by flow cytometry
The expression frequency of the upper Cx43 of CD8+ is respectively (0.14 ± 0.01) %, (0.26 ± 0.11) %, (0.24 ± 0.08) %, SHR group
It expresses and rises compared with WKY rat group, no significant difference, NaHS intervention group is expressed compared with SHR group to be declined, no statistical difference meaning
Justice, Fig. 8.
The foregoing is only a preferred embodiment of the present invention, the scope of protection of the present invention is not limited to this, it is any ripe
Know those skilled in the art within the technical scope of the present disclosure, the letter for the technical solution that can be become apparent to
Altered or equivalence replacement are fallen within the protection scope of the present invention.
Claims (1)
1. a kind of experimental method for proving hydrogen sulfide and gap connection relationship, which comprises the following steps:
Grouping: WKY group: normal Wistar capital of a country, rat, male and female are regardless of, 9 week old, daily intraperitoneal injection and NaHS processing group etc.
Physiological saline is measured, continues 8 weeks；
SHR group: spontaneous hypertensive rat, male and female are regardless of, 9 week old, daily intraperitoneal injection and NaHS processing group normal salt
Water continues 8 weeks；
NaHS processing group: spontaneous hypertensive rat, male and female are regardless of, 9 week old, daily 56 μm of ol/kg weight of intraperitoneal injection
NaHS continues 8 weeks；
Step 1, animal blood pressure measurement
Each group rat tail artery systolic pressure is detected using full-automatic rats and mice non-invasive blood pressure measuring instrument；It is first preheated when measurement, by blood
The inflation tail sleeve of pressure meter covers in rat root of the tail portion, after rat is quiet, duplicate measurements blood pressure 3 times, as a result takes its average value；
Step 2, the preparation of Peripheral Lymphocytes of Rat suspension
Under rat anesthesia state, each group rat aorta peripheral blood is acquired, is placed in 10g/LEDTA vacuum blood collection tube；Outside
All blood is centrifuged, and serum and haemocyte are separated, and few one times of PBS solution is added in haemocyte, is mixed well, will be dilute
Haemocyte after releasing gently is laid on equivalent lymphocyte separation medium, 1500r, 30min, extracts cloud in 1.5ml centrifuge tube,
1800r, 6min abandon supernatant, and PBS solution is resuspended, spare；
The detection of step 3, T lymphocyte subsets and Cx40, Cx43
Streaming special dyeing pipe is taken, each group rat is respectively labeled as blank tube, measurement pipe and Isotype control pipe in sequence, each to manage
It is separately added into corresponding streaming antibody, adjusts antibody final concentration according to antibody specification, and set up negative control；Room temperature is protected from light incubation
30min uses on the machine CELLQuest software and is detected；It marks and 120 μ L of fixative is added in the pipe of CD4-APC and CD8-PE,
4 DEG C of incubation 30min, then with rupture of membranes buffer rupture of membranes, according to specification reference concentration be added goat anti-rat Cx40 antibody and
The goat anti-mouse of the rabbit-anti goat IgG of FITC label, FITC Isotype control and small anti-rat Cx43 antibody and FITC label
IgG, FITC Isotype control, 37 DEG C of incubation 30min, use on the machine CELLQuest software and are detected；Flow cytometer is adjusted
To optimum Working, with FSC/SSC gating and dead cell and cell fragment are excluded, experiment parameter is adjusted according to Isotype control
Carry out sample measures again afterwards, gating first is in CD3+ lymphocyte, then lymph of the gating after CD4+ and CD8+ cell, rupture of membranes
Cell gating is in CD4+ and CD8+, and then for gating in Cx40+, Cx43+ cell, all sample standard deviations complete the inspection to cell in 2h
It surveys, every pipe detects 20000 cells, and computer reads measured value, finally analyzes the data of collection；
Corresponding streaming antibody specific described in step 3 are as follows: CD3-FITC, FITC Isotype control；CD4-APC, APC homotype pair
According to；CD8-PE, PE Isotype control；The rabbit-anti goat IgG of goat anti-rat Cx40 antibody and FITC label, FITC Isotype control；
The goat anti-mouse IgG of small anti-rat Cx43 antibody and FITC label, FITC Isotype control.
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| EP3889603A4 (en) * | 2018-12-01 | 2022-08-10 | Mingdao Innovation (Beijing) Medical-Tech Co., Ltd. | Preparation method for lymphocyte sample for flow cytometry analysis |
| CN111208306A (en) * | 2020-03-28 | 2020-05-29 | 安徽大学 | Method for measuring expression quantity of nicotinic acetylcholine receptor in rat lymphocyte |
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Non-Patent Citations (4)
| Title |
|---|
| Effect of Novel Gasotransmitter hydrogen sulfide on renal fibrosis and connexins expression in diabetic rats;Ou Zeng et al.;《Bioengineered》;20160830;第7卷(第5期);第314-320页 |
| 硫化氢对糖尿病大鼠心肌纤维化及CX40和CX45表达的影响;肖婷 等;《中国老年学杂志》;20151130;第35卷(第21期);摘要，第6025-6026页"1 材料与方法" |
| 自发性高血压大鼠外周血CD4+/CD8+T淋巴细胞比率增加且连接子蛋白40(Cx40)和炎症因子水平升高;王爱 等;《细胞与分子免疫学杂志》;20160229;第32卷(第2期);第145-148页 |
| 自发性高血压大鼠肾损伤伴随调节性T淋巴细胞下降及连接蛋白43的表达增强;张海超 等;《免疫学杂志》;20151130;第31卷(第11期);第941-944页 |
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