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Файл:Ординатура / Хирургия / Библиотека им академика М.И. Перельмана / Книга_1649_Библиотеки_им_академика_М_И_Перельмана.pdf
Beneficial Effect and Underlying Mechanism …
201
PAF and NF-B Decreased after SMI Treatment
According to the Lab test results, we observed that much less amylase was
released to serum from pancreas after ginsenoside treatment and hypothesized
it results from the alleviated inflammatory reaction. PAF was detected in
serum via ELISA, and similar with the trend of amylase, the PAF expression
decreased gradually after the induction of pancreatitis. Moreover, the PAF of
SMI group at d3, d5 and d7 were significantly lower than counterparts in
control (P < 0.01) (Table 1). In addition to PAF, a significantly lower mRNA
and protein level of NF-B was expressed at d7 and d14 (both P < 0.05
respectively) (Table 2, Figure 1) in pancreas tissue in SMI group via real-time
quantitative PCR, compared with control group. PAF and NF-B are
inflammation-related factors. Then the hypothesis was confirmed that SMI
attenuated the inflammation of acute pancreatitis and alleviated the
progressive damage and promoted the recovery of pathological pancreas.
Table 2. Comparison of NF-κB and CD31 expression between
the two groups
Group
Number
Days post induction of pancreatitis
7d
14d
NF-κB
SMI
Control
30
30
0.83±0.03
*
1.00±0.06
0.44±0.02
*
0.76±0.05
CD31 positive
expression SMI
Control
30
30
6.41±1.14
**
3.62±0.89
9.03±2.55
**
3.22±1.92
*
P < 0.05,
**
P < 0.01, vs control.
Figure 1. NF-κB activity and β-actin expression of the pancreas by Western blot Test.
Qiang Yan, Wei Wu, Xing Yao et al.
202
Figure 2. Mean MVD of the pancreas at d7 and d14 for the two groups. A) pancreas
collected at d7 of the SMI group; B) pancreas collected at d14 of the SMI group; C)
pancreas collected at d7 of the control group; D) pancreas collected at d14 of the
control group; E) pancreas at the time of induction of pancreatitis.
SMI Promoted Angiogenesis and Improved Microcirculation
Differing from PAF and NF-B, VEGF and AF increased steadily with
the pancreatitis. Interestingly, both VEGF and AF of SMI group at d3, d5 and
d7 were significantly higher than those in control (P < 0.05) (Table 1). Both
VEGF and AF are associated with angiogenesis and establishment of
microcirculation. CD31, involved in angiogenesis, was expressed significantly
higher in SMI group both at d7 and d14 (P < 0.01 respectively) (Table 2).
Besides, the mean rate of MVD as assessed by CD31 was higher in SMI group
compared to control in IHC assay (Figure 2). Thus, SMI promoted
Beneficial Effect and Underlying Mechanism …
203
angiogenesis during the recovery of acute pancreatitis, improved
microcirculation establishment and finally alleviated the inflammation reaction
and improved the prognosis.
DISCUSSION
Studies have demonstrated that the occurrence of acute pancreatitis was
associated with pancreatic self-digestion of trypsin, microcirculation
disturbance, destruction of the pancreatic duct barrier, oxygen free radicals,
inflammatory mediators and a variety of cytokines. However, cytokine
network and immune dysfunction maybe the main reasons to precipitate the
rapid progress of pancreatitis from local lesions to systemic inflammatory
response syndrome (SIRS) and multiple organ dysfunction [8]. Although to
suppress trypsinogen activation was believed to be the primary treatment for
patients with acute pancreatitis or severe acute pancreatitis in the past years,
ways to improve microcirculation had currently been adopted to mitigate cell
damage and necrosis and consequently improve or prevent general
microcirculatory disturbance and multiple organ dysfunction given that
microcirculation dysfunction had been one of the main mechanism for the
pancreatitis [9].
AF is a large family of polypeptide which can promote angiogenesis under
physiological and pathological conditions, accelerate blood vessel maturation,
and maintenance of vascular stability. However, the VEGF is a purified class
of angiogenic factors. VEGF is a secretary protein and has been shown to only
stimulate endothelial cells and it can induce and promote angiogenesis in vivo.
PAF, a biologically active phospholipid, is expressed by blood cells, platelets,
endothelial cells, lung, liver, kidney, and other cell types. The factor was then
activated by binding the specific receptor in the target cell membrane. A large
of studies [10, 11] had demonstrated the association between the PAF and the
pathogenesis of pancreatitis. Potential mechanisms including: 1) PAF is able
to directly modulate microvascular permeability and increase venular
permeability. Increased capillary permeability permits sequestration of
macromolecules and fluid, which causes a deficiency of circulating blood
volume and microcirculatory disorders. Consequently, prolonged local
hypoxia or anoxia, and necrosis ensued. 2) PAF promotes the activation and
adhesion of platelets and may cause microthrombi formation, which can also
lead to the deterioration of pancreatic microcirculation and pancreatic necrosis.
3) PAF is shown to be involved in adhesion, chemotaxis, degranulation and,
Qiang Yan, Wei Wu, Xing Yao et al.
204
even the whole procedure of activation of leukocytes. Besides, it can amplify
inflammatory response. As a result, large amounts of oxygen free radicals
were released to the circulation and subsequently disintegrate the lysosome,
which then causes the cell damage and death. 4) Activation of phospholipase
A2 by PAF and mutual positive feedback further aggravates tissue injury. All
the above indicated that the higher the PAF is in the pancreatitis, the more
severe the disease is. SMI, a Chinese traditional prescription mainly consisting
of two herbal components, Radix ginseng Rubra and Radix ophiopogonis, is
extensively being used in China as a microcirculation protector because SMI
has been shown to protect endothelial cells and increase the vasodilators
released, which consequently dilate the vessels and amply the blood flow.
Moreover, SMI has been documented to activate the adrenal system and
enhance the ability of reticulo-endothelial system to scavenge various
pathological materials. As a result, the blood flows to vital organs like heart,
liver, brain were increased and the microcirculation was increased. In this
study, both VEGF and AF of SMI group at d3, d5 and d7 were significantly
higher than those in control (P < 0.05). This means the SMI treatment after
caerulein-induced pancreatitis may promote the angiogenesis and attenuate
inflammatory response. By contrast, PAF of amylase of SMI group at d3, d5
and d7 were significantly lower than counterparts in control, i.e., the
inflammatory response of pancreatitis was greatly mitigated after
administration of SMI.
The “excessive leukocyte activation” theory has attracted increasing
attentions for the occurrence of pancreatitis with more studies focused on the
pathogenesis of acute pancreatitis. Nevertheless, cytokines played a vital role
in the excessive leukocyte activation and the progress into systemic
inflammatory response syndrome. NF–κB is a multi-directional regulatory
protein, responsible for cytokine production and cell survival. It may modulate
gene expression in various other situations that signal rapid gene expression
and plays an important role in the immune regulation, inflammation, stress
response and apoptosis. Activated NF–κB could translocate from cytoplasm to
nucleus rapidly and bind to the κB domain of promoter or enhancer of the
target genes. This consequently regulate the transcription of factors including
cytokines such as TNF-α, IL-1β, IL-2 and IL-6, chemokines such as IL-8,
macrophage chemotactic peptides, adhesion molecules like E-selectin, growth
factors like IL-3, immunoglobulin light chain class I major hisocompatibility
complex. Therefore, the NF–κB if of paramount importance in the regulation
of inflammatory immune response given all the above factors would involve in
the inflammatory response [12, 13]. However, excessive activation of NF–κB
Beneficial Effect and Underlying Mechanism …
205
would enhance general immune response, which resulted in the tissue damage
and organ dysfunction. Telek and colleagues [14] demonstrated that the
activation of NF–κB was associated with early acinar oxidative stress and
consequently tissue damage in pancreatitis model. Further study by Hietaranta
et al., [15] shown that NF–κB activation per se is required for the
inflammatory exacerbation and extra-pancreatic organ injury such as acute
respiratory distress syndrome in the developing phase. Given the essential role
of reactive oxygen species in the activation of NF–κB, the antioxidants could
thereby used as NF–κB inhibitors. Coincidently, Kim et al., [16] revealed that
antioxidants such as N-acetylcysteine might be useful antiinflammatory agents
in acute pancreatitis mode by inhibiting oxidant-mediated activation of NF–κB
and decreasing cytokine production. The main ingredients for ShenMai Injection
are ginseng saponin, Radix saponins, flavonoids and trace Radix ginseng
polysaccharides and polysaccharides and have been shown to have a wide
variety of biological activities including immunomodulatory effect,
antioxidant, anti-inflammatory, and anti-tumor activity. As shown in this
study, the SMI significantly decreased the expression of NF–κB at d7 and d14
compared to the control. This implies the antioxidant effect of SMI may
mitigate the inflammation and enhance the survival.
CD31, also known as platelet-endothelial cell adhesion molecule
(
PECAM-1
)
is a member of the immunoglobulin gene superfamily. As a main
adhesive molecule of vascular endothelial cells, CD31 is mainly expressed in
endothelial cells, platelets, monocytes, neutrophils, naive CD8 + T cells and
NK cells [17]. As a new marker of microvessels, CD31 is superior to other
markers of endothelial cells to identify the vessels [18]. Thus, the CD31 could
be used as specific markers to differentiate the endothelial cells and
quantitative evaluation of the role of angiogenic factors played in the process
of angiogenesis [19].
MVD is a useful and common marker to reflect
angiogenesis. Given the CD31 is a main marker of vascular endothelial cells, it
plays an important role in angiogenesis and is expressed in both mature and
non-mature blood vessels. CD31 are expressed in the cytoplasm of endothelial
cells and brown staining, hence the MVD count and CD31
immunohistochemical analysis could accurately reflect the number of
microvessels [20]. In our study, more positive CD 31 were expressed in the
SMI group at d7 and d14 and so did mean MVD assessed by CD31 positive
staining. We hypothesized that the SMI may enhance CD31 positive
expression, increase MVD and prompt angiogenesis. Microcirculation was
consequently improved and this may attenuate the acute pancreatitis.
Qiang Yan, Wei Wu, Xing Yao et al.
206
In summary, SMI could promote angiogenesis and improve
microcirculation. Consequently, the inflammatory mediators were reduced and
inhibited, and the injury to pancreatic acinar cells was then greatly alleviated.
Hence, the SMI may be beneficial in the treatment of acute pancreatitis.
CONFLICTS OF INTEREST
None declared.
FUNDING
This study was partially supported by the Clinical Scientific Research of
project funds in Zhejiang Province Medical Association (No. 2009 zyc35)
REFERENCES
[1] Mitchell, R. M., M. F. Byrne, and J. Baillie, Pancreatitis. Lancet, 2003.
361(9367): p. 1447-55.
[2] Gloor, B., et al., Kupffer cell blockade reduces hepatic and systemic
cytokine levels and lung injury in hemorrhagic pancreatitis in rats.
Pancreas, 2000. 21(4): p. 414-20.
[3] Menger, M. D., T. Plusczyk, and B. Vollmar, Microcirculatory
derangements in acute pancreatitis. J Hepatobiliary Pancreat Surg,
2001. 8(3): p. 187-94.
[4] Wang, R. X., et al., Ginsenoside Rb1 attenuates agonist-induced
contractile response via inhibition of store-operated calcium entry in
pulmonary arteries of normal and pulmonary hypertensive rats. Cell
Physiol Biochem, 2015. 35(4): p. 1467-81.
[5] Zhou, Q., et al., Ginsenoside Rg1 inhibits platelet activation and arterial
thrombosis. Thromb Res, 2014. 133(1): p. 57-65.
[6] Zhou, W., et al., Ginsenoside Rb1 blocks homocysteine-induced
endothelial dysfunction in porcine coronary arteries. J Vasc Surg, 2005.
41(5): p. 861-8.
Beneficial Effect and Underlying Mechanism …
207
[7] Weidner, N., et al., Tumor angiogenesis: a new significant and
independent prognostic indicator in early-stage breast carcinoma. J Natl
Cancer Inst, 1992. 84(24): p. 1875-87.
[8] Pezzilli, R., Acute recurrent pancreatitis: an autoimmune disease?
World J Gastroenterol, 2008. 14(7): p. 999-1006.
[9] Schneider, L., et al., Protective effects and anti-inflammatory pathways
of exogenous calcitonin gene-related peptide in severe necrotizing
pancreatitis. Pancreatology, 2009. 9(5): p. 662-9.
[10] Formela, L. J., et al., Amelioration of experimental acute pancreatitis
with a potent platelet-activating factor antagonist. Br J Surg, 1994.
81(12): p. 1783-5.
[11] Chen, C., et al., Therapy for acute pancreatitis with platelet-activating
factor receptor antagonists. World J Gastroenterol, 2008. 14(30): p.
4735-8.
[12] Siebenlist, U., G. Franzoso, and K. Brown, Structure, regulation and
function of NF-kappa B. Annu Rev Cell Biol, 1994. 10: p. 405-55.
[13] Stancovski, I. and D. Baltimore, NF-kappaB activation: the I kappaB
kinase revealed? Cell, 1997. 91(3): p. 299-302.
[14] Telek, G., et al., Differential upregulation of cellular adhesion molecules
at the sites of oxidative stress in experimental acute pancreatitis. J Surg
Res, 2001. 96(1): p. 56-67.
[15] Hietaranta, A. J., et al., Relationship between NF-kappaB and
trypsinogen activation in rat pancreas after supramaximal caerulein
stimulation. Biochem Biophys Res Commun, 2001. 280(1): p. 388-95.
[16] Kim, H., et al., Suppression of NF-kappaB activation and cytokine
production by N-acetylcysteine in pancreatic acinar cells. Free Radic
Biol Med, 2000. 29(7): p. 674-83.
[17] Woodfin, A., M. B. Voisin, and S. Nourshargh, PECAM-1: a multi-
functional molecule in inflammation and vascular biology. Arterioscler
Thromb Vasc Biol, 2007. 27(12): p. 2514-23.
[18] Cristina, C., et al., VEGF and CD31 association in pituitary adenomas.
Endocr Pathol, 2010. 21(3): p. 154-60.
[19] Deepak, A. V. and B. P. Salimath, Antiangiogenic and proapoptotic
activity of a novel glycoprotein from U. indica is mediated by NF-
kappaB and Caspase activated DNase in ascites tumor model.
Biochimie, 2006. 88(3-4): p. 297-307.
[20] Sumpio, B. E., et al., MAPKs (ERK1/2, p38) and AKT can be
phosphorylated by shear stress independently of platelet endothelial cell
Qiang Yan, Wei Wu, Xing Yao et al.
208
adhesion molecule-1 (CD31) in vascular endothelial cells. J Biol Chem,
2005. 280(12): p. 11185-91.
BIOGRAPHICAL SKETCH
Name: Qiang Yan
Affiliation: Zhejiang University Huzhou hospital
Education: 09/1988-07/1993 Zhejiang Medical University, China.
Bachelor of Medicine. 09/2007-11/2013 Zhejiang University medical school,
China. MD, PhD
Address: #198 Hongqi Road, Huzhou, Zhejiang 313000, China.
Research and Professional Experience: Dr. Qiang Yan joined the
Department of Surgery in Zhejiang University Huzhou Hospital in 1997,
became the Director of the Division of HPB Surgery in 2009 and Chairman of
the Department of Surgery in 2015. Taking the special training at Department
of HPB and Minimally invasive Surgery Universität Regensburg Germany in
2009 and Stanford University USA in 2012, Dr. Yan has special experience in
advanced laparoscopic liver and pancreatic surgery.
Professional Appointments:
Chairman and Professor of Department of Surgery and Director of
Division of Hepatobiliary and pancreatic surgery, Zhejiang University Huzhou
hospital
Member of Gastrointestinal Physicians Branch of World endoscopy
Doctors Association, Member of Chinese college of Surgeons, Chinese
Medical Doctor Association,
Member of the standing committee, College of Surgeons, Zhejiang
Medical Doctor Association Member of division of Enteral parenteral
nutrition, Chinese Medical Association Zhejiang Branch.
Invited reviewer of journal of Medicine of Johns Hopkins Hospital,
Member of editor board of some Chinese medical journals.
Honors: 2005, awarded the third prize of science and technology
progress, Huzhou City, Zhejiang Province, China.
2006, awarded the third prize of medical science and technology progress,
Zhejiang Province, China.
2009, awarded the Youth Science and Technology Award of Huzhou City
Beneficial Effect and Underlying Mechanism …
209
2013, awarded the third prize of medical science and technology progress,
Zhejiang Province, China.
Publications Last 3 Years:
1. The management of injuries of choledocbo-pancreatico-duodenal
junction in Biliary operation. J Surg Con Prac, 2015, 20(3): 199-204. (In
Chinese).
2. Effects of timing of enteral nutrition in the severe acute pancreatitis.
Chin J Dig Surg, 2015, 14(5): 390-4. (In Chinese).
3. The impact of antiviral therapy on postoperative intrahepatic recurrence
of hepatitis B related hepatocellular carcinoma, Chin J Hepatobiliary
Surg, 2015, 21(2)91-5. (In Chinese).
4. Augmented miR-10b expression associated with depressed expression of
its target gene KLF4 involved in gastric carcinoma. International journal
of clinical and experimental pathology 2015, 8(5):5071-9.
5. Clinical application value of laparoscopic intraoperative
cholangiography in laparoscopic cholecystectomy. Chin J Prim Pharm,
2015, 22(7):996-8. (In Chinese).
6. Therapeutic effect of Shenmai injection on acute pancreatitis and
mechanism, Chin J Pancreatol, 2014, 14(4): 255-8. (In Chinese).
7. Clinical application value of laparoscopic treatment of mesenteric cyst.
Zh J J Traumatic. 2014, 19(6): 919-20. (In Chinese).
8. Three Points on Laparoscope Operation to Gallbladder Neck
Incarceration Stone. Journal of Zhejiang University of Traditional
Chinese Medicine, 2013, 37(11): 1319-21. (In Chinese).
9. Efficacy of postoperative antiviral combined transcatheter arterial
chemoembolization therapy in prevention of hepatitis B-related
hepatocellular. Chinese medical journal, 2013, 126(5) 855-9.
10. The Associated Ion between the VDR Gene Polymorphisms and
Susceptibility to Hepatocellular Carcinoma and the Clinicopathological
Features in Subjects Infected with HBV. Biomed Res Int, 2013, 953-74.
11. Prevention and Control System of Hypokalemia in Fast Recovery After
Abdominal Surgery. Current Therapeutic Research, 2013(3), 74:68–73.
12. Novel functional proteins interact with midkine in hepatic cancer cells.
HBPD INT 2012, 11(3):272-7.
13. Expression of FHIT, FN and PTEN in hepatocellular carcinomas. Chin J
Gen Surg, 2012, 27(2):487-90. (In Chinese).
Qiang Yan, Wei Wu, Xing Yao et al.
210
14. Clinicopathological and prognostic significance of epithelial
mesenchymal transition-related protein expression in intrahepatic
cholangiocarcinoma. OncoTargets and Therapy 2012, 5(10):255-61.
15. Effects of combined enteral nutrition support on hemorrheologic
parameters and the level of inflammatory factors in rabbits with severe
acute pancreatitis. Zhonghua yi xue za zhi, 2011, 91(28):2006-10. (In
Chinese).