Diabetes Research and Clinical Practice - Volume 120 - Supplement 1

Morisky-4 Questionnaire was completed for every patient

during the telephone interview. The end point of this study

is to evaluate the clinical biomarker such as A1C, Fasting

glucose, Cholesterol, LDL-C to determine the disease control

status objectively and the effects of pharmaceutical care. Data

was assessed by comparing baseline characteristics using

independent sample t-tests for continuous variables and chi-

square tests for categorical variables. Outcomes for continuous

variables were using paired t-tests to study the effect within

each study group. Data was run by the Excel

software.

Result:

From Oct. 2013 to May 2015, 76 eligible patients were

enrolled and informed consent were obtained. The patients

were divided into two groups by their A1C. There were 37

patients in the WC group, and 30 patients in the PC group.

Patients

’

characteristics (gender, age, education et al.) were not

statistically different between the two groups, except the

duration of disease (WC vs PC, 5.9 y vs 9.4 y p = 0.002). The

results of questionnaire showed that the patient

’

s recognition

of the importance of medication adherence was improved,

however, this acknowledgement had not been revealed by the

difference of their laboratory data.

Discussion:

Although, the results of patient

’

s clinical out-

come was not as improved as we expected. We found that

their awareness of being actively engaged in glycemic con-

trol was much better with intensive caregiver intervention.

We expected to establish a communication model for

physicians, nurses, pharmacists, and all other health care

providers to achieve and implement an individualized holistic

patient care.

Beta Cell Replacement Therapy

and Imaging

PH-01

Different dose of PSP/reg affect biological behavior of beta-cell

through induction of autophagy and apoptosis

Juan CHEN

*, Ying WANG

, Wei LI

, Wei XU

, Zilin SUN

Department of Endocrinology, Zhongda Hospital, Institute of

Diabetes, Medical School, Southeast University, China

Background and aims:

A great deal of evidence indicates

that pancreatic stone protein/regenerating protein (PSP/reg)

promotes beta-cell proliferation and the administration of PSP/

reg may be advantageous to improve diabetes. The aim of this

study was to investigate whether PSP/reg provide protective

effect only within certain concentration range and the

underlying mechanism.

Materials and methods:

MIN-6 cells were treated with or

without PSP/reg at different dose (0, 30, 100, 200, 300 ng/mL).

Cell viability was measured by CCK-8 assay, cell function was

determined by measuring insulin secretion, autophagy level

was measured by detecting LC3-II, LC3-I and p62 expression,

cell apotosis was measured by flow cytometry.

Result:

MIN-6 cells viability maintained optimum state when

the dose reached to 100 ng/mL and incubated for 24 h (P < 0.01).

However, high dose of PSP/reg (300 ng/mL) resulted in a

significant decrease in cells viability. No significant difference

in insulin secretion was observed at the dose of 30 and 0 ng/

mL. But insulin secretion significantly increased at 100 ng/mL

and decreased at 300 ng/mL. Autophagy level significantly

elevated as determined by the increased ratio of LC3-II to LC3-I

and decreased p62 expression, a dose-dependent manner in

this effect was observed. Therewas no significant difference in

the rate of apoptosis between treatment with PSP/reg (100 ng/

mL) and the control group.

Conclusion:

In the present study, low concentrations of

PSP/reg is able to preserve the viability and function of MIN-6

cells, but high concentrations of PSP/reg can exert cellular

toxicity. These evidence leads us to conclude that treatment

with PSP/reg at an appropriate concentration is necessary to

induce protective effect, the mechanism may be related to

autophagy and apoptosis.

PH-02

Involvement of glycosphingolipids in the insulin secretion

pathway

Iwao TAKAHASHI

*, Wakana MATSUYAMA

, Shinji GO

Jin-ichi INOKUCHI

, Koji NATA

Department of Medical

Biochemistry, School of Pharmacy, Iwate Medical University, Yahaba,

Division of Glycopathology, Institute of Molecular Biomembrane and

Glycobiology, Tohoku Medical and Pharmaceutical University,

Sendai, Japan

Background and aims:

Glycosphingolipids (GSLs) are amphi-

pathic molecules consisting of a hydrophilic sugar chain head

and a hydrophobic ceramide tail. They are mostly located in

the outer leaflet of the plasma membrane, in which they are

anchored by the ceramide moiety. Their sugar chain moieties

have enormous structural diversity, and the variety of glycan

structures in GSLs contributes to determine specific cell

functions. As for involvement of GSLs in the field of diabetes

study, roles of GSLs in diabetic insulin resistance are well

known. However, the contribution of GSLs to insulin secretion

-cell function is still unclear. The aim of this study is to

investigate whether the GSLs in

-cells affects insulin

secretion.

Materials andmethods:

MIN6 cells, mouse pancreatic beta-cell

line, have been reported to show functional heterogeneity with

increasing passage number. Therefore, for analysis of the

relationship of the GSLs and ß-cell function, selection of a

single optimal cell line was required. Subcloning of MIN6 cells

was performed by the limiting dilutionmethod. The cells were

then screened and selected by an index of glucose- and

potassium-stimulated insulin secretion (GSIS and KSIS). The

glycosphingolipids in MIN6 and its isolated subclone cells

were analysed by thin-layer chromatography. Insulin content,

GSIS and KSIS were examined in MIN6 subclones after

inhibition of the first enzyme for GSLs biosynthesis by D-

PDMP (a competitive inhibitor of glucosylceramide synthase).

Quantitative RT-PCR was used for analyzing mRNA expression

of components of insulin secretion pathway.

Results:

MIN6 and GSIS and/or KSIS responsible subclone cells

commonly expressed GM3 and GD1a gangliosides. Subcloned

MIN6 cells treated with D-PDMP showed about 82.8%, 81.1%

and 56.6% decrease in insulin content, GSIS and KSIS

compared to the controls (p < 0.01), respectively. In addition,

the mRNA level of Insulin1, Glut2, Sur1 and Snap25 were

decreased in D-PDMP-treated subcloned MIN6 cells when

compared with the control cells (p < 0.01). Therefore, the

decreased mRNA expression of these genes may have

reduced the insulin content and secretory response in the

D-PDMP-treated cells.

Conclusion:

These results indicate that GSLs, especially

GM3 and/or GD1a, play important role(s) in insulin secre-

tion mechanisms by influencing not only the insulin biosyn-

thesis but also the insulin secretion pathway. The reduction of

KSIS, which stimulates insulin secretion by directly depolar-

izing the plasma membrane, was decreased than that of GSIS,

suggesting that GSLs play a role in insulin secretion by

affecting the pathway downstream of ATP production.

PH-03

Cytomegalovirus transmission after islet transplant in

comparison to other solid organ transplant

Kitty Kit-Ting CHEUNG

*, Dima KABBANI

, Jutta PREIKSAITIS

Curtis MABILANGAN

, Richard ORAM

, James SHAPIRO

Peter SENIOR

Clinical Islet Transplant Program, University of

Alberta,

Provincial Laboratory for Public Health, University of

Alberta, Canada

Poster Presentations / Diabetes Research and Clinical Practice 120S1 (2016) S65

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